Anti-6X His tag® antibody [EPR20547] - ChIP Grade

40 product images

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  Immunoprecipitation - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204)

  His-tagged Staphylococcus aureus cas9 was immunoprecipitated from 0.35 mg of HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) transfected with a His-tagged Staphylococcus aureus cas9 (J7RUA5; aa1-1053; 125 kDa) construct, whole cell lysate with ab213204 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab213204 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366), was used for detection at 1/10,000 dilution
  

  Lane 1: HEK-293T transfected with His-tagged Staphylococcus aureus cas9 construct, whole cell lysate 10 μg (Input).

  Lane 2: ab213204 IP in HEK-293T transfected with His-tagged Staphylococcus aureus cas9 construct, whole cell lysate.

  Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab213204 in HEK-293T transfected with His-tagged Staphylococcus aureus cas9 construct, whole cell lysate.

  Blocking/Dilution buffer: 5% NFDM/TBST.

  Exposure time: 1 second.

  All lanes: Immunoprecipitation - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204)

  Observed band size: 125 kDa

• 

  ChIP - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204)

  Chromatin was prepared from MCF7 (human breast adenocarcinoma cell line) cells transfected with 6X His-tagged GATA3 according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10 minutes. The ChIP was performed with 25μg of chromatin, 2μg of ab213204 (blue), and 20μl of A/G sepharose beads slurry (10μl of sepharose A beads + 10μl of sepharose G beads). 2μg of rabbit normal IgG was added to the beads control (yellow). The immunoprecipitated DNA was quantified by real time PCR (Sybr green approach).
  

  ChIP was performed according to the literature (PMID:22951069).

• 

  Immunocytochemistry/ Immunofluorescence - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204)

  Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) cells transfected with GFP-Myc-His vector expression construct labeling 6X His tag® with ab213204 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody at 1/1000 dilution (white). Confocal image showing positive staining on HEK-293T cells transfected with GFP-Myc-His vector expression construct.
  

  The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) (Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889) (red) at 1/200 dilution.

  Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody at 1/1000 dilution.

• 

  Flow Cytometry - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204)

  Flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol permeabilized HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) transfected with GFP-Myc-His vector labeling 6X His tag® with ab213204 (right panel) at 1/5000 dilution compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) (left panel). Goat Anti-Rabbit IgG H&L (Alexa Fluor® 647) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 647) ab150079) at 1/2000 dilution was used as the secondary antibody.
  

  Gate is set between transfected and untransfected HEK-293T cells.

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204)

  Immunohistochemical analysis of agarose-embedded HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) transfected with a His-tagged Staphylococcus aureus cas9 (J7RUA5; aa1-1053; 125kDa) construct labeling 6X His tag® with ab213204 at 1/16000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use.
  

  Left image: Positive staining on HEK-293T transfected with a His-tagged Staphylococcus aureus cas9 (J7RUA5; aa1-1053; 125kDa) construct. Right image: No staining on HEK-293T transfected with an empty expression vector.

  Counter stained with Hematoxylin.

  Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

  Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204)

  Immunohistochemical analysis of paraffin-embedded human liver tissue labeling 6X His tag® with ab213204 at 1/16000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
  

  Negative control: No staining on human liver.

  Counter stained with Hematoxylin.

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204)

  Immunohistochemical analysis of paraffin-embedded rat stomach tissue labeling 6X His tag® with ab213204 at 1/16000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
  

  Negative control: No staining on rat stomach.

  Counter stained with Hematoxylin.

• 

  Western blot - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204)

  Blocking/Dilution buffer: 5% NFDM/TBST.

  All lanes: Western blot - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204) at 1/20000 dilution

  Lane 1: His-tagged mouse Cathepsin D recombinant protein (aa65-310; 27kDa) at 0.02 µg

  Lane 2: His-tagged mouse CDKN2A/p16INK4a recombinant protein (aa1-168;18kDa) at 0.02 µg

  Secondary

  All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution

  Developed using the ECL technique.

  Observed band size: 18 kDa, 27 kDa

  Exposure time: 1s

• 

  Western blot - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204)

  Blocking/Dilution buffer: 5% NFDM/TBST.
  

  Exposure time: Lane 1-3: 1 second; Lane 4-5: 10 seconds.

  All lanes: Western blot - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204) at 1/5000 dilution

  Lane 1: HEK-293T (human epithelial cell from embryonic kidney transformed with large T antigen) transfected with an empty expression vector, whole cell lysate at 20 µg

  Lane 2: HEK-293T (human epithelial cell from embryonic kidney transformed with large T antigen) transfected with a His-tagged Staphylococcus aureus cas9 (J7RUA5; aa1-1053;125kDa) construct, whole cell lysate at 20 µg

  Lane 3: HEK-293T (human epithelial cell from embryonic kidney transformed with large T antigen) transfected with a His-tagged human Brachyury (O15178; aa1-435;50kDa) construct, whole cell lysate at 20 µg

  Lane 4: HEK-293T (human epithelial cell from embryonic kidney transformed with large T antigen) transfected with a His-tagged human PRDM9 (Q9NQV7; aa1-894;100kDa) construct, whole cell lysate at 20 µg

  Lane 5: HEK-293T (human epithelial cell from embryonic kidney transformed with large T antigen) transfected with a His-tagged human GAPDH (P04406; aa1- 335;36kDa) construct, whole cell lysate at 20 µg

  Secondary

  All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution

  Developed using the ECL technique.

  Observed band size: 100 kDa, 125 kDa, 36 kDa, 50 kDa

• 

  Western blot - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204)

  6X His tag® Western blot staining using rabbit Anti-6X His tag® antibody

  In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) staining at 1/200000 dilution.

  In Western blot, Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204) staining at 1/5000 dilution.

  All lanes: Western blot - Anti-Aggrecan antibody [RM2036] (Anti-Aggrecan antibody [RM2036] ab315486) at 1/1000 dilution

  Lane 1: 293T cells transfected with an empty vector containing a His-tag, whole cell lysate at 20 µg

  Lane 2: 293T cells transfected with a Human ACAN expression vector containing a His-tag, whole cell lysate at 20 µg

  Secondary

  All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution

  Observed band size: 75 kDa, 36 kDa

  Exposure time: 1s

• 

  Western blot - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204)

  6X His tag® Western blot staining using rabbit Anti-6X His tag® antibody

  Blocking and diluting buffer and concentration: 5% NFDM/TBST.

  This antibody does not cross-react with human MAP2 and MAP4.

  In Western blot, Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204) staining at 1/5000 dilution.

  All lanes: Western blot - Anti-Tau (N-terminal) antibody [EPR29521-8] (Anti-Tau (N-terminal) antibody [EPR29521-8] ab324236) at 1/1000 dilution

  Lane 1: 293T cells transfected with an empty vector containing a myc-His-tag®, whole cell lysate at 20 µg

  Lane 2: 293T cells transfected with a human Tau 2N4R expression vector containing a myc-His-tag®, whole cell lysate at 20 µg

  Lane 3: 293T cells transfected with a human PNS-Tau expression vector containing a DDDDK-tag, whole cell lysate at 20 µg

  Lane 4: 293T cells transfected with a human MAP2 expression vector containing a DDDDK-tag, whole cell lysate at 20 µg

  Lane 5: 293T cells transfected with a human MAP4 expression vector containing a DDDDK-tag, whole cell lysate at 20 µg

  Secondary

  All lanes: Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/100000 dilution

  Observed band size: 33-60 kDa, 15-150 kDa, 37-250 kDa, 36 kDa

  Exposure time: 1s

• 

  Western blot - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204)

  6X His tag® Western blot staining using rabbit Anti-6X His tag® antibody

  Blocking and diluting buffer and concentration: 5% NFDM/TBST.

  This antibody does not cross-react with recombinant human PACSIN2 and PACSIN3 by Western Blot.

  In Western blot, Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204) staining at 1/5000 dilution.

  All lanes: Western blot - Anti-PACSIN1 antibody [EPR29589-587] (Anti-PACSIN1 antibody [EPR29589-587] ab323573) at 1/1000 dilution

  Lane 1: No-tagged human PACSIN1 recombinant protein at 20 ng

  Lane 2: His-tagged human PACSIN2 recombinant protein at 20 ng

  Lane 3: His-tagged human PACSIN3 recombinant protein at 20 ng

  Secondary

  All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution

  Observed band size: 51 kDa, 50-55 kDa

  Exposure time: 180s

• 

  Western blot - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204)

  6X His tag® Western blot staining using rabbit Anti-6X His tag® antibody

  Blocking and diluting buffer and concentration: 5% NFDM/TBST

  Anti-Histone H3 antibody [EPR16987] - Nuclear Marker and ChIP Grade (Anti-Histone H3 antibody [EPR16987] - Nuclear Marker and ChIP Grade ab176842) was used as loading control at a 1/100000 dilution.

  Anti-6X His tag^® antibody [EPR20547] - ChIP Grade (ab213204) was used as total protein control at a 1/5000 dilution.

  All lanes: Western blot - Anti-HIV1 p24 antibody [24-2] (Anti-HIV1 p24 antibody [24-2] ab85536) at 1/1000 dilution

  Lane 1: 293T (human embryonic kidney epithelial cell) cells transfected with an empty vector containing a myc-His-tag® whole cell lysate at 4 µg

  Lane 2: 293T cells transfected with a Human immunodeficiency virus type 1 group M subtype B (isolate HXB2) (HIV-1) expression vector containing a myc-His-tag® whole cell lysate at 4 µg

  Lane 3: 293T cells transfected with a Human immunodeficiency virus type 1 group M subtype D (isolate NDK) (HIV-1) expression vector containing a myc-His-tag® whole cell lysate at 4 µg

  Lane 4: 293T cells transfected with a Human immunodeficiency virus type 1 group M subtype B (isolate WMJ22) (HIV-1) expression vector containing a myc-His-tag® whole cell lysate at 4 µg

  Lane 5: 293T cells transfected with a Human immunodeficiency virus type 1 group M subtype K (isolate 96CM-MP535) (HIV-1) expression vector containing a myc-His-tag® whole cell lysate at 4 µg

  Secondary

  All lanes: Peroxidase-Conjugated Goat anti-Mouse IgG (H+L) at 1/10000 dilution

  Observed band size: 28 kDa

  Exposure time: 3s

• 

  Western blot - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204)

  6X His tag® Western blot staining using rabbit Anti-6X His tag® antibody

  Blocking and diluting buffer and concentration: 1% BSA/TBST.

  Anti-Vinculin antibody [EPR8185] (Anti-Vinculin antibody [EPR8185] - Loading Control ab129002) (1/10000) (124KDa); Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204) (1/5000);

  In Western blot, Anti-STING antibody [EPR13130-55] (Anti-STING antibody [EPR13130-55] ab239074) staining at 1/1000 dilution.

  All lanes: Western blot - Anti-STING (phospho S366) antibody [EPR29040-93] (Anti-STING (phospho S366) antibody [EPR29040-93] ab324229) at 1/1000 dilution

  Lane 1: 293T (human embryonic kidney epithelial cell) cells transfected with an empty vector containing a myc-His-tag®, whole cell lysate at 20 µg

  Lane 2: 293T cells transfected with a human wild-type STING expression vector containing a myc-His-tag®, whole cell lysate at 20 µg

  Lane 3: 293T cells transfected with a human wild-type STING expression vector containing a myc-His-tag® treated first with 80nM TPA for 24h, then change fresh medium, transfect 10ug/ml poly(dA:dT) and treated with 10 uM MG-132 for 6 hours, 100 uM Calyculin A was then added for additional 30 minutes, whole cell lysate at 20 µg

  Lane 4: 293T cells transfected with a human STING (S366A mutation) expression vector containing a myc-His-tag®, whole cell lysate at 20 µg

  Lane 5: 293T cells transfected with a human STING (S366A mutation) expression vector containing a myc-His-tag® treated first with 80nM TPA for 24h, then change fresh medium, transfect 10ug/ml poly(dA:dT) and treated with 10 uM MG-132 for 6 hours, 100 uM Calyculin A was then added for additional 30 minutes, whole cell lysate at 20 µg

  Secondary

  All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution

  Observed band size: 40 kDa, 124 kDa

  Exposure time: 59s

• 

  Dot Blot - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204)

  6X His tag® Dot Blot staining using rabbit Anti-6X His tag® antibody

  Dot blot analysis of SFRP2 using Anti-SFRP2 antibody [EPR26088-157] ab318185 at 1:1000 (0.501 ug/ml) followed by a Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1:100,000 dilution.

  Blocking and diluting buffer and concentration: 5% NFDM/TBST.

  Lane 1: His-tagged human SFRP2 recombinant protein
  
  Lane 2: His-tagged human SFRP1 recombinant protein

  Anti-6X His tag ® antibody [EPR20547] - ChIP Grade (ab213204) (1:1000) as total protein control.

  This antibody does not cross-react with human SFRP1.
  

  All lanes: Dot Blot - Anti-SFRP2 antibody [EPR26088-157] (Anti-SFRP2 antibody [EPR26088-157] ab318185) at 1/1000 dilution

  Lane 1: His-tagged human SFRP2 recombinant protein

  Lane 2: His-tagged human SFRP1 recombinant protein

  Secondary

  All lanes: Dot Blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution

  Exposure time: 180s

• 

  Dot Blot - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204)

  6X His tag® Dot Blot staining using rabbit Anti-6X His tag® antibody

  Dot blot analysis of LARP1 using Anti-LARP1 antibody [EPR28797-92] ab317575 at 1:1000 (0.519 ug/ml) followed by a Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1:100,000 dilution.

  Blocking and diluting buffer and concentration: 5% NFDM/TBST.

  This antibody does not cross-react with human LARP1B.

  All lanes: Dot Blot - Anti-LARP1 antibody [EPR28797-92] (Anti-LARP1 antibody [EPR28797-92] ab317575) at 1/1000 dilution

  Lane 1: His-tagged human LARP1 fragment

  Lane 2: His-tagged human LARP1B fragment

  Secondary

  All lanes: Dot Blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution

  Exposure time: 180s

• 

  Western blot - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204)

  6X His tag® Western blot staining using rabbit Anti-6X His tag® antibody

  This antibody does not cross-react with mouse Syntenin-2.

  In Western blot, Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204) staining at 1/5000 dilution.

  All lanes: Western blot - Anti-Syntenin antibody [EPR28604-40] (Anti-Syntenin antibody [EPR28604-40] ab315342) at 1/1000 dilution

  Lane 1: His-tagged mouse Syntenin-1 recombinant protein at 10 ng

  Lane 2: His-tagged mouse Syntenin-2 recombinant protein at 10 ng

  Secondary

  All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution

  Observed band size: 35 kDa

  Exposure time: 180s

• 

  Western blot - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204)

  6X His tag® Western blot staining using rabbit Anti-6X His tag® antibody

  Blocking and diluting buffer and concentration: 5% NFDM/TBST.

  The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID: 15123664, PMID: 8421459).

  In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) staining at 1/200000 dilution.

  In Western blot, Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204) staining at 1/5000 dilution.

  All lanes: Western blot - Anti-Surfactant Protein A/PSAP antibody [EPR29097-290] (Anti-Surfactant Protein A/PSAP antibody [EPR29097-290] ab322404) at 1/1000 dilution

  Lane 1: 293T (human embryonic kidney epithelial cell) cells transfected with an empty vector containing a His-tag whole cell lysate at 20 µg

  Lane 2: 293T cells transfected with a mouse SP-A expression vector containing a His-tag, whole cell lysate at 20 µg

  Secondary

  All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution

  Observed band size: 35 kDa, 70 kDa, 124 kDa

  Exposure time: 3s

• 

  Dot Blot - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204)

  6X His tag® Dot Blot staining using rabbit Anti-6X His tag® antibody

  Dot blot analysis of CD55 using Anti-CD55 antibody [EPR28962-549] ab324067 at 1:1000 (0.489 ug/ml) followed by a Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1:100,000 dilution.

  Lane 1: 293T cells transfected with a mouse CD55 expression vector containing a myc-His-tag®, whole cell lysate
  
  Lane 2: 293T cells transfected with a mouse DAF2 expression vector containing a myc-His-tag®, whole cell lysate

  Exposure time: 180 seconds.

  Blocking and diluting buffer and concentration: 5% NFDM/TBST.

  This antibody does not cross-react with mouse DAF2 .

  In Dot Blot, Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204) staining at 1/5000 dilution.

  All lanes: Dot Blot - Anti-CD55 antibody [EPR28962-549] (Anti-CD55 antibody [EPR28962-549] ab324067) at 1/1000 dilution

  Lane 1: 293T cells transfected with a mouse CD55 expression vector containing a myc-His-tag®, whole cell lysate

  Lane 2: 293T cells transfected with a mouse DAF2 expression vector containing a myc-His-tag®, whole cell lysate

  Secondary

  All lanes: Dot Blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution

  Exposure time: 180s

• 

  Western blot - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204)

  6X His tag® Western blot staining using rabbit Anti-6X His tag® antibody

  This antibody does not cross-react with human beta Synuclein and alpha Synuclein.

  In Western blot, Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204) staining at 1/5000 dilution.

  In Western blot, Anti-alpha + beta Synuclein antibody [EP1646Y] - (Anti-alpha + beta Synuclein antibody [EP1646Y] ab51252) staining at 1/1000 dilution.

  All lanes: Western blot - Anti-gamma Synuclein/SNCG antibody [EPR26218-79] (Anti-gamma Synuclein/SNCG antibody [EPR26218-79] ab315442) at 1/1000 dilution

  Lane 1: His-tagged human gamma Synuclein recombinant protein at 10 ng

  Lane 2: His-tagged human beta Synuclein recombinant protein at 10 ng

  Lane 3: His-tagged human alpha Synuclein recombinant protein at 10 ng

  Secondary

  All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution

  Observed band size: 14 kDa, 20 kDa, 19 kDa

  Exposure time: 59s

• 

  Western blot - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204)

  6X His tag® Western blot staining using rabbit Anti-6X His tag® antibody

  Blocking and diluting buffer and concentration: 5% NFDM/TBST
  This antibody does not cross-react with human STAG2 (isoform-1).
  In Western blot, Anti-Vinculin antibody [EPR8185] (Anti-Vinculin antibody [EPR8185] - Loading Control ab129002) staining at 1/10000 dilution.
  In Western blot, Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204) staining at 1/5000 dilution
  Exposure time: 180 seconds

  All lanes: Western blot - Anti-SA2 (isoform-2) antibody [EPR26892-61] (Anti-SA2 (isoform-2) antibody [EPR26892-61] ab313877) at 1/1000 dilution

  Lane 1: 293T cells transfected with an empty vector containing a His tag, whole cell lysate at 20 µg

  Lane 2: 293T cells transfected with a human STAG2 (isoform-1) expression vector containing a His-tag, whole cell lysate at 20 µg

  Lane 3: 293T cells transfected with a human STAG2 (isoform-2) expression vector containing a His-tag, whole cell lysate  at 20 µg

  Secondary

  All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution

  Observed band size: 25-30 kDa, 150 kDa

  Exposure time: 180s

• 

  Western blot - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204)

  6X His tag® Western blot staining using rabbit Anti-6X His tag® antibody

  This antibody does not cross-react with overexpressed Flagellin (Salmonella anatum), Flagellin (Salmonella typhimurium (strain LT2 /SGSC1412 / ATCC 700720)) and Flagellin (Salmonella paratyphi A (strain ATCC9150 / SARB42)).

  In Western blot, Anti-Vinculin antibody [EPR8185] (Anti-Vinculin antibody [EPR8185] - Loading Control ab129002) staining at 1/10000 dilution.

  In Western blot, Anti-6X His tag antibody [EPR20547]-CHIP Grade (ab213204) staining at 1/5000 dilution.

  All lanes: Western blot - Anti-Flagellin antibody [EPR28356-51] (Anti-Flagellin antibody [EPR28356-51] ab316010) at 1/1000 dilution

  Lane 1: 293T cells transfected with a Flagellin (E.coli K12) expression vector containing a His-tag, whole cell lysate at 20 µg with 5% NFDM/TBST

  Lane 2: 293T cells transfected with a Flagellin (Salmonella anatum) expression vector containing a His-tag, whole cell lysate at 20 µg with 5% NFDM/TBST

  Lane 3: 293T cells transfected with a Flagellin (Salmonella typhimurium (strain LT2 / SGSC1412 / ATCC 700720)) expression vector containing a His-tag, whole cell lysate at 20 µg with 5% NFDM/TBST

  Lane 4: 293T cells transfected with a Flagellin (Salmonella paratyphi A (strain ATCC 9150 / SARB42)) expression vector containing a His-tag, whole cell lysate at 20 µg with 5% NFDM/TBST

  Secondary

  All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution

  Observed band size: 51 kDa, 124 kDa

  Exposure time: 15s

• 

  Western blot - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204)

  6X His tag® Western blot staining using rabbit Anti-6X His tag® antibody

  Rfx Cas13d may cleave to two fragments close to C-term, and this antibody can recognize the full length and N-terminal fragment. His-tag antibody can recognize full length and C-terminal fragment because the His-tag of this overexpression vector located on C-term.

  In Western blot, Anti-Vinculin antibody [EPR8185] (Anti-Vinculin antibody [EPR8185] - Loading Control ab129002) staining at 1/10000 dilution.

  In Western blot, Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204) staining at 1/5000 dilution.

  All lanes: Western blot - Anti-Rfx Cas13d (N-term) antibody [EPR23965-31] (Anti-Rfx Cas13d (N-term) antibody [EPR23965-31] ab314741) at 1/1000 dilution

  Lane 1: 293T (human embryonic kidney epithelial cell) cells transfected with an empty vector containing a myc-His-tag®, whole cell lysate at 10 µg with 5% NFDM/TBST

  Lane 2: 293T cells transfected with a Ruminococcus flavefaciens cas13d expression vector containing a myc-His-tag® (C-term), whole cell lysate at 10 µg with 5% NFDM/TBST

  Secondary

  All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution

  Observed band size: 110 kDa, 70 kDa, 124 kDa, 40 kDa

  Exposure time: 6s

• 

  Western blot - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204)

  6X His tag® Western blot staining using rabbit Anti-6X His tag® antibody

  In Western blot, Anti-Vinculin antibody [EPR8185] (Anti-Vinculin antibody [EPR8185] - Loading Control ab129002) staining at 1/10000 dilution.

  In Western blot, Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204) staining at 1/5000 dilution.

  All lanes: Western blot - Anti-Hepatitis B Virus Core Antigen antibody [EPR28251-34] (Anti-Hepatitis B Virus Core Antigen antibody [EPR28251-34] ab316283) at 1/1000 dilution

  Lane 1: 293T cells (human embryonic kidney epithelial cell) transfected with an empty vector containing a myc-His-tag®, whole cell lysate at 20 µg with 5% NFDM/TBST

  Lane 2: 293T cells transfected with a Virus wild-type Hepatitis B Virus Core Antigen expression vector containing a myc-His-tag®, whole cell lysate at 20 µg with 5% NFDM/TBST

  Secondary

  All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution

  Observed band size: 25 kDa, 124 kDa

  Exposure time: 6s

• 

  Western blot - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204)

  6X His tag® Western blot staining using rabbit Anti-6X His tag® antibody

  Blocking and diluting buffer and concentration: 5% NFDM/TBST.
  
  This antibody does not cross-react with human UCK1.
  
  In Western blot, anti-His antibody (ab213204) staining at 1/5000 dilution.
  
  In Western blot, anti- Vinculin antibody (Anti-Vinculin antibody [EPR8185] - Loading Control ab129002) loading control staining at 1/10000 dilution.
  
  Exposure time: 6 seconds.

  All lanes: Western blot - Anti-UCK2 antibody [EPR27020-82] (Anti-UCK2 antibody [EPR27020-82] ab308252) at 1/1000 dilution

  Lane 1: 293T cells transfected with an empty vector containing a His tag whole cell lysate at 20 µg

  Lane 2: 293T cells transfected with a Human UCK2 expression vector containing a His-tag whole cell lysate at 20 µg

  Lane 3: 293T cells transfected with a Human UCK1 expression vector containing a His-tag whole cell lysate at 20 µg

  Secondary

  All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution

  Observed band size: 30 kDa

• 

  Western blot - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204)

  6X His tag® Western blot staining using rabbit Anti-6X His tag® antibody

  All lanes: Western blot - Anti-CUX1+CUX2 antibody [EPR26509-154] (Anti-CUX1+CUX2 antibody [EPR26509-154] ab309139) at 1/1000 dilution

  Lane 1: 293T cells transfected with an empty vector containi a His-tag whole cell lysate at 20 µg

  Lane 2: 293T cells transfected with a human CUX1 expression vector containi a His-tag whole cell lysate at 20 µg

  Lane 3: 293T cells transfected with a human CUX2 expression vector containi a His-tag whole cell lysate at 20 µg

  Secondary

  All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution

  Observed band size: 200 kDa

  Exposure time: 6s

• 

  Western blot - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204)

  6X His tag® Western blot staining using rabbit Anti-6X His tag® antibody

  Blocking and diluting buffer and concentration: 5% NFDM/TBST

  The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID: 18649180; PMID: 30448479).

  This blot was developed using a high sensitivity ECL substrate. The high-sensitivity ECL substrate used allows for the detection of proteins in the mid-femtogram range.

  Alkaline phosphatase. -: untreated membrane; +: phosphatase treated membrane.

  In Western blot, anti-His antibody (ab213204) staining at 1/5000 dilution.

  Anti-GAPDH antibody (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) loading control staining at 1/200000 dilution.

  Anti-Connexin 43 / GJA1 antibody [EPR21153] (Anti-Connexin 43 / GJA1 antibody [EPR21153] - Intercellular Junction Marker ab217676) total protein control staining at 1/1000 dilution.

  All lanes: Western blot - Anti-Connexin 43 / GJA1 (phospho Y265) antibody [EPR26416-134] (Anti-Connexin 43 / GJA1 (phospho Y265) antibody [EPR26416-134] ab312836) at 1/1000 dilution

  Lane 1: HeLa (human cervical adenocarcinoma epithelial cell) cells transfected with an empty vector containi a myc-His-tag® whole cell lysate at 20 µg

  Lane 2: HeLa cells transfected with a human GJA1 expression vector containi a myc-His-tag® whole cell lysate at 20 µg

  Lane 3: HeLa cells transfected with a human GJA1(Mutant Y265F) expression vector containi a myc-His-tag® whole cell lysate at 20 µg

  Lane 4: HeLa cells transfected with a RSV-SR-A V-SRC expression vector containi a myc-His-tag® whole cell lysate at 20 µg

  Lane 5: HeLa cells co-transfected with a human GJA1 expression vector containi a myc-His-tag® and a RSV-SR-A V-SRC expression vector containi a myc-His-tag® whole cell lysate at 20 µg

  Lane 6: HeLa cells co-transfected with a human GJA1(Mutant Y265F) expression vector containi a myc-His-tag® and a RSV-SR-A V-SRC expression vector containi a myc-His-tag® whole cell lysate at 20 µg

  Secondary

  All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution

  Developed using the ECL technique.

  Observed band size: 43 kDa

  Exposure time: 92s

• 

  Western blot - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204)

  6X His tag® Western blot staining using rabbit Anti-6X His tag® antibody

  Blocking and diluting buffer and concentration: 5% NFDM/TBST
  In Western blot, anti- Vinculin antibody (Anti-Vinculin antibody [EPR8185] - Loading Control ab129002) loading control staining at 1/10000 dilution.
  In Western blot, anti-His antibody (ab213204) staining at 1/5000 dilution.
  Exposure time: 6 seconds

  All lanes: Western blot - Anti-SYT13 antibody [EPR26923-67] (Anti-SYT13 antibody [EPR26923-67] ab313329) at 1/1000 dilution

  Lane 1: 293T cells transfected with an empty vector containi a His-tag whole cell lysate at 20 µg

  Lane 2: 293T cells transfected with a Human SYT13 expression vector containi a His-tag whole cell lysate at 20 µg

  Secondary

  All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution

  Observed band size: 55 kDa

  Exposure time: 6s

• 

  Western blot - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204)

  6X His tag® Western blot staining using rabbit Anti-6X His tag® antibody

  This antibody does not cross-react with human PLXNB2 and human PLXNB3.

  His tag not visible in empty vector control image as out of range due to small size

  In Western blot, Anti-Vinculin antibody [EPR8185] (Anti-Vinculin antibody [EPR8185] - Loading Control ab129002) staining at 1/10000 dilution.

  In Western blot, Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204) staining at 1/5000 dilution.

  All lanes: Western blot - Anti-Plexin B1 antibody [EPR26309-65] (Anti-Plexin B1 antibody [EPR26309-65] ab316312) at 1/1000 dilution

  Lane 1: 293T (human embryonic kidney epithelial cell) cells transfected with an empty vector containing a His-tag, whole cell lysate at 20 µg with 5% NFDM/TBST

  Lane 2: 293T cells transfected with a PLXNB1 expression vector containing a His-tag, whole cell lysate at 20 µg with 5% NFDM/TBST

  Lane 3: 293T cells transfected with a PLXNB2 expression vector containing a His-tag, whole cell lysate at 20 µg with 5% NFDM/TBST

  Lane 4: 293T cells transfected with a PLXNB3 expression vector containing a His-tag, whole cell lysate at 20 µg with 5% NFDM/TBST

  Secondary

  All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution

  Observed band size: 75-300 kDa, 124 kDa

  Exposure time: 70s

• 

  Western blot - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204)

  6X His tag® Western blot staining using rabbit Anti-6X His tag® antibody

  Blocking and diluting buffer and concentration: 5% NFDM/TBST.

  The identity of the lower MW band at approximately 190 kDa and higher MW band at approximately 460 kDa are unknown.

  In Western blot, Anti-Vinculin antibody [EPR8185] (Anti-Vinculin antibody [EPR8185] - Loading Control ab129002) staining at 1/10000 dilution.

  In Western blot, Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204) staining at 1/5000 dilution.

  All lanes: Western blot - Anti-ATM (phospho S1987) antibody [EPR28058-71] (Anti-ATM (phospho S1987) antibody [EPR28058-71] ab315019) at 1/1000 dilution

  Lane 1: 293T cells transfected with an empty vector containing a myc-His-tag®, whole cell lysate at 20 µg

  Lane 2: 293T cells transfected with a mouse wild-type ATM expression vector containing a myc-His-tag®, whole cell lysate at 20 µg

  Lane 3: 293T cells transfected with a mouse ATM (S1987A mutation) expression vector containing a myc-His-tag®, whole cell lysate at 20 µg

  Secondary

  All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution

  Observed band size: 349 kDa, 124 kDa

  Exposure time: 180s

• 

  Dot Blot - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204)

  6X His tag® Dot Blot staining using rabbit Anti-6X His tag® antibody

  Dot blot analysis of Perilipin-2 using Anti-Perilipin-2 antibody [EPR28638-74] ab318134 at 1:1000 (0.524 ug/ml) followed by a Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1:100,000 dilution.

  Blocking and diluting buffer and concentration: 5% NFDM/TBST.

  This antibody does not cross-react with human Perilipin-3 and Perilipin-5.

  Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204) (1:1000) as total protein control.

  All lanes: Dot Blot - Anti-Perilipin-2 antibody [EPR28638-74] (Anti-Perilipin-2 antibody [EPR28638-74] ab318134) at 1/1000 dilution

  Lane 1: His-tagged human Perilipin-2 recombinant protein

  Lane 2: His-tagged human Perilipin-5 recombinant protein

  Lane 3: His-tagged human Perilipin-3 recombinant protein

  Secondary

  All lanes: Dot Blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution

  Exposure time: 10s

• 

  Dot Blot - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204)

  6X His tag® Dot Blot staining using rabbit Anti-6X His tag® antibody

  Dot blot analysis of Nav1.7 using Anti-Nav1.7 antibody [EPR26246-318] ab323849 at 1:1000 (0.493 ug/ml) followed by a Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1:100,000 dilution.

  Lane1: 293T cells transfected with a mouse SCN2A expression vector containing a His-tag, whole cell lysate
  
  Lane2: 293T cells transfected with a mouse SCN3A expression vector containing a His-tag, whole cell lysate
  
  Lane3: 293T cells transfected with a mouse SCN1A expression vector containing a His-tag, whole cell lysate
  
  Lane4: 293T cells transfected with a mouse SCN9A expression vector containing a His-tag, whole cell lysate

  Exposure time: 180 seconds.

  Blocking and diluting buffer and concentration: 5% NFDM/TBST.

  This antibody does not cross-react with mouse SCN1A, SCN2A and SCN3A.

  In Dot Blot, Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204) staining at 1/5000 dilution.

  All lanes: Dot Blot - Anti-Nav1.7 antibody [EPR26246-318] (Anti-Nav1.7 antibody [EPR26246-318] ab323849) at 1/1000 dilution

  Lane 1: 293T cells transfected with a mouse SCN2A expression vector containing a His-tag, whole cell lysate

  Lane 2: 293T cells transfected with a mouse SCN3A expression vector containing a His-tag, whole cell lysate

  Lane 3: 293T cells transfected with a mouse SCN1A expression vector containing a His-tag, whole cell lysate

  Lane 4: 293T cells transfected with a mouse SCN9A expression vector containing a His-tag, whole cell lysate

  Secondary

  All lanes: Dot Blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution

  Exposure time: 180s

• 

  Western blot - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204)

  6X His tag® Western blot staining using rabbit Anti-6X His tag® antibody

  This antibody does not cross-react with mouse Cyclin E2.

  In Western blot, Anti-Vinculin antibody [EPR8185] (Anti-Vinculin antibody [EPR8185] - Loading Control ab129002) staining at 1/10000 dilution.

  In Western blot, Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204) staining at 1/5000 dilution.

  All lanes: Western blot - Anti-Cyclin E1 antibody [EPR25394-23] (Anti-Cyclin E1 antibody [EPR25394-23] ab317436) at 1/1000 dilution

  Lane 1: 293T cells transfected with an empty vector containing a His-tag, whole cell lysate at 20 µg with NFDM/TBST

  Lane 2: 293T cells transfected with a mouse Cyclin E1 expression vector containing a His-tag, whole cell lysate at 20 µg with NFDM/TBST

  Lane 3: 293T cells transfected with a mouse Cyclin E2 expression vector containing a His-tag, whole cell lysate at 20 µg with NFDM/TBST

  Secondary

  All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution

  Observed band size: 50 kDa, 124 kDa

  Exposure time: 10s

• 

  Western blot - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204)

  6X His tag® Western blot staining using rabbit Anti-6X His tag® antibody

  In Western blot, Anti-Vinculin antibody [EPR8185] (Anti-Vinculin antibody [EPR8185] - Loading Control ab129002) staining at 1/10000 dilution.

  In Western blot, Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204) staining at 1/5000 dilution.

  All lanes: Western blot - Anti-POLE (mutated P286R) antibody [EPR28640-56] (Anti-POLE (mutated P286R) antibody [EPR28640-56] ab318136) at 1/1000 dilution

  Lane 1: 293T cells transfected with an empty vector containing a myc-His-tag®, whole cell lysate at 20 µg with NFDM/TBST

  Lane 2: 293T cells transfected with a human wild-type POLE expression vector containi a myc-His-tag®, whole cell lysate at 20 µg with NFDM/TBST

  Lane 3: 293T cells transfected with a human POLE (mutated P286R) expression vector containi a myc-His-tag®, whole cell lysate at 20 µg with NFDM/TBST

  Secondary

  All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution

  Observed band size: 261 kDa, 124 kDa

  Exposure time: 180s

• 

  Western blot - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204)

  6X His tag® Western blot staining using rabbit Anti-6X His tag® antibody

  Blocking and diluting buffer and concentration: Intercept® (TBS) Blocking Buffer diluted with an equal volume of TBS.

  The samples were run on a Bis-Tris gel under reducing conditions.

  Western blot: Anti-Alpha-S1-casein antibody (Anti-Alpha-S1-casein antibody [EPR29093-719] ab323188) staining at 1/1000 dilution, shown in green(up); Mouse anti-GAPDH antibody 6C5 loading control staining at 1/20000 dilution, shown in magenta; Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204) at 1/5000 dilution, shown in green(down).

  Milk Powder Trim was diluted using 1%TBS buffer.

  Avoid using milk-based buffers for blocking and dilution, as they might cause interference.

  In Western blot, Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204) staining at 1/5000 dilution.

  All lanes: Western blot - Anti-Alpha-S1-casein antibody [EPR29093-719] (Anti-Alpha-S1-casein antibody [EPR29093-719] ab323188) at 1/1000 dilution

  Lane 1: 293T (human embryonic kidney epithelial cell) cells transfected with an empty vector containing a myc-His-tag®, whole cell lysate at 20 µg

  Lane 2: 293T cells transfected with a Bovine Alpha-S1-casein expression vector containing a myc-His-tag®, whole cell lysate at 20 µg

  Secondary

  Lanes 1 - 2: Goat Anti-Rabbit IgG H&L (800CW) at 1/100000 dilution

  Lanes 1 - 2: Goat Anti-Mouse IgG H&L (680RD) at 1/100000 dilution

  Performed under reducing conditions.

  Observed band size: 34 kDa, 36 kDa

  Exposure time: 180s

• 

  Western blot - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204)

  6X His tag® Western blot staining using rabbit Anti-6X His tag® antibody

  In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) staining at 1/200000 dilution.

  In Western blot, Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204) staining at 1/5000 dilution.

  All lanes: Western blot - Anti-gamma Synuclein/SNCG antibody [EPR26218-79] (Anti-gamma Synuclein/SNCG antibody [EPR26218-79] ab315442) at 1/1000 dilution

  Lane 1: 293T cells transfected with an empty vector containing a myc-His-tag®, whole cell lysate at 5 µg

  Lane 2: 293T cells transfected with a human gamma Synuclein expression vector containing a myc-His-tag®, whole cell lysate at 5 µg

  Lane 3: 293T cells transfected with a mouse gamma Synuclein expression vector containing a myc-His-tag®, whole cell lysate at 5 µg

  Secondary

  All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution

  Observed band size: 17 kDa, 19 kDa, 36 kDa

  Exposure time: 6s

• 

  Western blot - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204)

  6X His tag® Western blot staining using rabbit Anti-6X His tag® antibody

  In Western blot, Anti-Vinculin antibody [EPR8185] (Anti-Vinculin antibody [EPR8185] - Loading Control ab129002) staining at 1/10000 dilution.

  In Western blot, Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204) staining at 1/5000 dilution.

  All lanes: Western blot - Anti-HERV-K-env antibody [EPR27172-7] (Anti-HERV-K-env antibody [EPR27172-7] ab319031) at 1/1000 dilution

  Lane 1: 293T (human embryonic kidney epithelial cell) cells transfected with an empty vector containing a myc-His-tag®, whole cell lysate at 20 µg with NFDM/TBST

  Lane 2: 293T cells transfected with a human HERV-K-env expression vector containing a myc-His-tag®, whole cell lysate at 20 µg with NFDM/TBST

  Secondary

  All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution

  Observed band size: 75 kDa, 124 kDa

  Exposure time: 15s

• 

  Western blot - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204)

  6X His tag® Western blot staining using rabbit Anti-6X His tag® antibody

  Blocking and diluting buffer and concentration: 5% NFDM/TBST.

  This antibody does weak cross-react with Bacillus subtilis RecA, but does not cross-react with Staphylococcus aureus RecA and Helicobacter pylori RecA .

  In Western blot, Anti-Vinculin antibody [EPR8185] (Anti-Vinculin antibody [EPR8185] - Loading Control ab129002) staining at 1/10000 dilution.

  In Western blot, Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204) staining at 1/5000 dilution.

  All lanes: Western blot - Anti-RecA antibody [EPR29094-561] (Anti-RecA antibody [EPR29094-561] ab323376) at 1/1000 dilution

  Lane 1: 293T (human embryonic kidney epithelial cell) cells transfected with an empty vector containing a His-tag, whole cell lysate at 20 µg

  Lane 2: 293T cells transfected with a E.coli RecA expression vector containing a His-tag, whole cell lysate at 20 µg

  Lane 3: 293T cells transfected with a Bacillus subtilis RecA expression vector containing a His-tag, whole cell lysate at 20 µg

  Lane 4: 293T cells transfected with a Staphylococcus aureus RecA expression vector containing a His-tag, whole cell lysate at 20 µg

  Lane 5: 293T cells transfected with a Helicobacter pylori RecA expression vector containing a His-tag, whole cell lysate at 20 µg

  Secondary

  All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution

  Observed band size: 35-45 kDa, 124 kDa

  Exposure time: 15s

• 

  Western blot - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204)

  6X His tag® Western blot staining using rabbit Anti-6X His tag® antibody

  Blocking and diluting buffer and concentration: 5% NFDM/TBST.

  In Western blot, Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204) staining at 1/5000 dilution.

  In Western blot, Anti-Vinculin antibody [EPR8185] (Anti-Vinculin antibody [EPR8185] - Loading Control ab129002) staining at 1/10000 dilution.

  In Western blot, Anti-Rb antibody [EPR17512-291] (Anti-Rb antibody [EPR17512] ab181616) staining at 1/1000 dilution.

  All lanes: Western blot - Anti-Rb (phospho S795) antibody [EPR28156-6] (Anti-Rb (phospho S795) antibody [EPR28156-6] ab321974) at 1/1000 dilution

  Lane 1: 293T cells transfected with a human Rb expression vector containing a His-tag, whole cell lysate at 20 µg

  Lane 2: 293T cells transfected with a human Rb (S795A mutation) expression vector containing a His-tag, whole cell lysate at 20 µg

  Secondary

  All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution

  Observed band size: 106 kDa, 124 kDa

  Exposure time: 1s

• 

  Western blot - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204)

  6X His tag® Western blot staining using rabbit Anti-6X His tag® antibody

  In Western blot, Anti-Vinculin antibody [EPR8185] (Anti-Vinculin antibody [EPR8185] - Loading Control ab129002) staining at 1/10000 dilution.

  In Western blot, Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204) staining at 1/5000 dilution.

  All lanes: Western blot - Anti-SCN11A antibody [EPR28603-66] (Anti-SCN11A antibody [EPR28603-66] ab316324) at 1/1000 dilution

  Lane 1: 293T cells transfected with an empty vector containing a His-tag, whole cell lysate at 10 µg with 5% NFDM/TBST

  Lane 2: 293T (human embryonic kidney epithelial cell) cells transfected with human SCN11A expression vector containi a myc-His-tag®, whole cell lysate at 10 µg with 5% NFDM/TBST

  Secondary

  All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution

  Observed band size: 204 kDa, 124 kDa

  Exposure time: 3s