Rabbit IgG, monoclonal [EPR25A] - Isotype Control

41 product images

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  Flow Cytometry - Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)

  Overlay histogram showing A549 (human lung carcinoma) cells stained with Anti-Integrin alpha 2 antibody [EPR5788] ab133557 (red line). The cells were fixed with 2% paraformaldehyde. The cells were then incubated with Anti-Integrin alpha 2 antibody [EPR5788] ab133557 at 1/60 dilution. The secondary antibody used was goat anti-rabbit IgG (FITC) at 1/150 dilution. Isotype control antibody (black line) was rabbit IgG (monoclonal) (ab172730). Unlabelled control (blue line) was cells without incubation with primary antibody and secondary antibody. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 530/30 bandpass filter.
  

  Conjugated versions are available for this clone: Alexa Fluor^® 488 (Alexa Fluor® 488 Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab199091), Alexa Fluor^® 647 (Alexa Fluor® 647 Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab199093), R-PE (PE Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab209478), APC (APC Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab232814).

• 

  Immunoprecipitation - Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)

  All lanes: Immunoprecipitation - Anti-IgA antibody [EPR5367-76] (Anti-IgA antibody [EPR5367-76] ab124716) at 1/1000 dilution

  Lane 1: Human plasma

  Lane 2: Anti-IgA antibody [EPR5367-76] ab124716 at 1/30 IP in Human plasma

  Lane 3: Rabbit monoclonal IgG (ab172730) instead of Anti-IgA antibody [EPR5367-76] ab124716 in Human plasma

  Secondary

  All lanes: Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1500 dilution

  Predicted band size: 83 kDa

  Observed band size: 60 kDa

• 

  Immunoprecipitation - Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)

  Anti-ATF2 (phospho T71) antibody [E268] ab32019 Immunoprecipitating ATF2 in Human Hela whole cell lysate. For western blotting Anti-ATF2 (phospho T71) antibody [E268] ab32019 (1:1000) was used to confirm successful immunoprecipitation. Blocking and diluting buffer used was 5% NFDM/TBST.

  All lanes: Immunoprecipitation - Anti-ATF2 (phospho T71) antibody [E268] (Anti-ATF2 (phospho T71) antibody [E268] ab32019) at 0.47 µg/mL

  Lane 1: HeLa (human cervix adenocarcinoma) treated with 250ng/ml anisomycin for 30min whole cell lysate (input) at 10 µg

  Lane 2: HeLa treated with 250ng/ml anisomycin for 30min whole cell lysate (+)

  Lane 3: Rabbit monoclonal IgG (ab172730) instead of Anti-ATF2 (phospho T71) antibody [E268] ab32019 in HeLa treated with 250ng/ml anisomycin for 30min whole cell lysate (-)

  Secondary

  All lanes: Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) at 1/10000 dilution

  Predicted band size: 55 kDa

  Exposure time: 3min

• 

  Immunoprecipitation - Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)

  SLAMF6 was immunoprecipitated from 0.35 mg Jurkat (human T cell leukemia cell line from peripheral blood) whole cell lysate with Anti-SLAMF6 antibody [EPR22170] ab224201 at 1/30 dilution. Western blot was performed from the immunoprecipitate using Anti-SLAMF6 antibody [EPR22170] ab224201 at 1/500 dilution. VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366), was used for detection at 1/1000 dilution.
  

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

  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.

  All lanes: Immunoprecipitation - Anti-SLAMF6 antibody [EPR22170] (Anti-SLAMF6 antibody [EPR22170] ab224201) at 1/500 dilution

  Lane 1: Jurkat whole cell lysate (input) at 10 µg

  Lane 2: Anti-SLAMF6 antibody [EPR22170] ab224201 at 1/30 IP in Jurkat whole cell lysate (+) at 10 µg

  Lane 3: Rabbit monoclonal IgG (ab172730) instead of Anti-SLAMF6 antibody [EPR22170] ab224201 in Jurkat whole cell lysate (-) at 10 µg

  Secondary

  All lanes: Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) at 1/1000 dilution

  Developed using the ECL technique.

  Predicted band size: 37 kDa

  Exposure time: 10s

• 

  Immunoprecipitation - Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)

  SLAMF6 was immunoprecipitated from 0.35 mg Ramos (human Burkitt's lymphoma cell line) whole cell lysate with Anti-SLAMF6 antibody [EPR22170] ab224201 at 1/30 dilution. Western blot was performed from the immunoprecipitate using Anti-SLAMF6 antibody [EPR22170] ab224201 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) was used for detection at 1/5000 dilution.
  

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

  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.

  All lanes: Immunoprecipitation - Anti-SLAMF6 antibody [EPR22170] (Anti-SLAMF6 antibody [EPR22170] ab224201) at 1/1000 dilution

  Lane 1: Ramos whole cell lysate 10 μg (Input) at 10 µg

  Lane 2: Anti-SLAMF6 antibody [EPR22170] ab224201 at 1/30 IP in Ramos whole cell lysate (+) at 10 µg

  Lane 3: Rabbit monoclonal IgG (ab172730) instead of Anti-SLAMF6 antibody [EPR22170] ab224201 in Ramos whole cell lysate (-) at 10 µg

  Secondary

  All lanes: Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) at 1/5000 dilution

  Developed using the ECL technique.

  Predicted band size: 37 kDa

  Exposure time: 10s

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  Immunoprecipitation - Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)

  Anti-IL-1RA antibody [EPR6483] ab124962 (purified) at 1/20 immunoprecipitating IL-1RA in NIH/3T3 whole cell lysate.
  Lane 1 (input): NIH/3T3 whole cell lysate (10μg)
  Lane 2 (+): Anti-IL-1RA antibody [EPR6483] ab124962 + NIH/3T3 whole cell lysate.
  Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of Anti-IL-1RA antibody [EPR6483] ab124962 in NIH/3T3 whole cell lysate.
  For western blotting, VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366), was used for detection at 1/10,000 dilution.
  Blocking buffer and concentration: 5% NFDM/TBST.
  Diluting buffer and concentration: 5% NFDM /TBST.

  All lanes: Immunoprecipitation - Anti-IL-1RA antibody [EPR6483] (Anti-IL-1RA antibody [EPR6483] ab124962)

  All lanes: NIH/3T3 whole cell lysate

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  Flow Cytometry - Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)

  Overlay histogram showing K562 (human chronic myelogenous leukemia) cells stained with ab196018 (red line). The cells were fixed with 2% paraformaldehyde. The cells were then incubated with ab196018 at 1/150 dilution. The secondary antibody used was goat anti-rabbit IgG (FITC) at 1/150 dilution. Isotype control antibody (black line) was rabbit IgG (monoclonal) (ab172730). Unlabelled control (blue line) was cells without incubation with primary antibody and secondary antibody. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 530/30 bandpass filter.

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  Immunoprecipitation - Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)

  Anti-Histone H4 (acetyl K8) antibody [EP1002Y] - ChIP Grade ab45166 (purified) at 1/20 immunoprecipitating Histone H4 (acetyl K8) in HeLa treated with Trichostatin A whole cell lysate.
  

  For western blotting, VeriBlot for IP Detection Reagent (HRP) ab131366 VeriBlot for IP Detection Reagent (HRP) was used for detection (1/10000).

  Blocking buffer and concentration: 5% NFDM/TBST.

  Diluting buffer and concentration: 5% NFDM /TBST.

  All lanes: Immunoprecipitation - Anti-Histone H4 (acetyl K8) antibody [EP1002Y] - ChIP Grade (Anti-Histone H4 (acetyl K8) antibody [EP1002Y] - ChIP Grade ab45166) at 1/20 dilution

  Lane 1: HeLa (human cervix adenocarcinoma) treated with Trichostatin A whole cell lysate at 10 µg

  Lane 2: (+) Anti-Histone H4 (acetyl K8) antibody [EP1002Y] - ChIP Grade ab45166 + HeLa treated with Trichostatin A whole cell lysate

  Lane 3: (-) Rabbit monoclonal IgG (ab172730) instead of Anti-Histone H4 (acetyl K8) antibody [EP1002Y] - ChIP Grade ab45166 in HeLa treated with Trichostatin A whole cell lysate.

  Secondary

  All lanes: Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) at 1/10000 dilution

  Predicted band size: 11 kDa

  Observed band size: 11 kDa

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  Immunoprecipitation - Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)

  LINE-1 ORF1p was immunoprecipitated from 0.35 mg F9 (mouse embryonal carcinoma epithelial cell) whole cell lysate with Anti-LINE-1 ORF1p antibody [EPR21844-108] ab216324 at 1/30 dilution. Western blot was performed from the immunoprecipitate using Anti-LINE-1 ORF1p antibody [EPR21844-108] ab216324 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366), was used for detection at 1/5000 dilution.
  

  Lane 1: F9 whole cell lysate 10 μg (Input).
  Lane 2: Anti-LINE-1 ORF1p antibody [EPR21844-108] ab216324 IP in F9 whole cell lysate.
  Lane 3: Rabbit monoclonal IgG (ab172730) instead of Anti-LINE-1 ORF1p antibody [EPR21844-108] ab216324 in F9 whole cell lysate.

  Blocking/Dilution buffer: 5% NFDM/TBST.
  Exposure time: 30 seconds.

  All lanes: Immunoprecipitation - Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)

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  Flow Cytometry - Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)

  Flow Cytometry staining using rabbit Anti- antibody

  Flow cytometric analysis of HEK-293T (Human embryonic kidney epithelial cell, Left panel) / Caco-2 (Human colorectal adenocarcinoma epithelial cell, Right panel) using Anti-CD133 antibody [RM1002] - Stem Cell Marker ab278053 at 1/500 dilution (0.1µg) (red). The isotype control used was the Rabbit monoclonal IgG (ab172730), black line and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (blue). Secondary antibody used was the Goat anti-rabbit IgG (Alexa Fluor^® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) at 1/2000 dilution.

  Negative control: HEK-293T. (PMID 20167130)
  Gated on viable cells.

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  Flow Cytometry - Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)

  Flow Cytometry staining using rabbit Anti- antibody

  Flow cytometric analysis of HEK-293T (Human embryonic kidney epithelial cell, Left)/ LNCaP (Human prostate carcinoma epithelial cell, Right) cells labelling TMPRSS2 with Anti-TMPRSS2 antibody [EPR24407-87] ab280567 at 1/500 dilution (0.1μg)/ red compared with Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat anti rabbit IgG (Alexa Fluor^® 647, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 647) ab150079) at 1/2000 dilution was used as the secondary antibody.
  

  Negative control: HEK-239T (PMID: 20631123).

  Gated on viable cells.

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  Flow Cytometry (Intracellular) - Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)

  Flow cytometry overlay histogram showing wild-type HAP1 (green line) and TP53 knockout HAP1 cells stained with Anti-p53 antibody [SP161] ab227655 (red line). The cells were fixed with 4% formaldehyde (10 min) and then permeabilized with 0.1% PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS containing 10% normal goat serum to block non-specific protein-protein interaction followed by the antibody (Anti-p53 antibody [SP161] ab227655) (1x10⁶ in 100 µl at 0.008 µg/ml) for 30 min at 22°C.
  

  The secondary antibody Goat anti-Rabbit IgG H&L (Alexa Fluor^® 488, pre-adsorbed) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) was used at 1/2000 for 30 min at 22°C.

  Isotype control antibody was Rabbit IgG (monoclonal) (ab172730) used at the same concentration and conditions as the primary antibody (wild-type HAP1 - black line TP53 knockout HAP1 - grey line). Unlabelled sample was also used as a control (this line is not shown for the purpose of simplicity).

  Acquisition of >5000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter.

  This antibody gave a positive signal in TP53 HAP1 knockout cells fixed with 80% methanol (5 min) / permeabilized with 0.1% PBS-Triton X-100 for 15 min used under the same conditions.

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  ChIC/CUT&RUN sequencing - Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)

  CUT&RUN was performed using the ChIC/CUT&RUN pAG-MNAse ChIC/CUT&RUN pAG-MNase ab285373 105 HeLa cells and 0.5 μg, 1 μg and 2μg of ab172730 [EPR25A]. H3K4me3 (Anti-Histone H3 (tri methyl K4) antibody [EPR20551-225] - ChIP Grade ab213224) and CTCF (Anti-CTCF antibody [EPR18253] - ChIP Grade ab188408) are used for comparison. The resulting DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 10 million reads
  Additional screenshots of mapped reads can be downloaded here.

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  Flow Cytometry - Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)

  Overlay histogram showing SH-SY5Y (human neuroblastoma) cells stained with Anti-beta Tubulin antibody [EPR16774] - Loading Control ab179513 (red line). The cells were fixed with 2% paraformaldehyde. The cells were then incubated with Anti-beta Tubulin antibody [EPR16774] - Loading Control ab179513 at 1/150 dilution. The secondary antibody used was goat anti-rabbit IgG (FITC) at 1/150 dilution. Isotype control antibody (black line) was rabbit IgG (monoclonal) (ab172730). Unlabelled control (blue line) was cells without incubation with primary antibody and secondary antibody. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 530/30 bandpass filter.

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)

  Immunohistochemical analysis of paraffin-embedded human colon tissue using anti-Vimentin RabMAb (Anti-Vimentin antibody [EPR3776] - Cytoskeleton Marker ab92547, left panel) (brown) and Rabbit mAb IgG control (ab172730, right panel).

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  Flow Cytometry - Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)

  Overlay histogram showing A549 (human lung carcinoma) cells stained with Anti-AKT1 + AKT2 + AKT3 antibody [EPR17671] ab185633 (red line). The cells were fixed with 2% paraformaldehyde. The cells were then incubated with Anti-AKT1 + AKT2 + AKT3 antibody [EPR17671] ab185633 at 1/150 dilution. The secondary antibody used was goat anti-rabbit IgG (FITC) at 1/150 dilution. Isotype control antibody (black line) was rabbit IgG (monoclonal) (ab172730). Unlabelled control (blue line) was cells without incubation with primary antibody and secondary antibody. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 530/30 bandpass filter

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)

  Immunohistochemical analysis of paraffin-embedded mouse lung tissue using anti-Vimentin RabMAb (Anti-Vimentin antibody [EPR3776] - Cytoskeleton Marker ab92547, left panel) (brown) and Rabbit mAb IgG control (ab172730, right panel).

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  Flow Cytometry (Intracellular) - Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)

  Intracellular flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized U-937 (Human histiocytic lymphoma monocyte) treated with 100ng/ml TPA for 24 hours, then 5μg/ml LPS for 4 hours, and add 300ng/ml BFA for another 3h (Red) / Untreated control (Green) cells labelling IL-8 with Anti-IL-8 antibody [EPR26511-74] ab289967 at 1/500 dilution (0.1μg) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor^® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 1/2000 dilution was used as the secondary antibody.

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human colon tissue with unpurified Rabbit IgG ab172730 at 1/10. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. A prediluted HRP-polymer conjugated anti-rabbit IgG was used as the secondary antibody. Counterstained with Hematoxylin.

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human colon tissue with purified Rabbit IgG ab172730 at 1/100. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. A prediluted HRP-polymer conjugated anti-rabbit IgG was used as the secondary antibody. Counterstained with Hematoxylin.

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  Flow Cytometry (Intracellular) - Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)

  Flow cytometric analysis of 4% paraformaldehyde fixed 0.1% Tween-20 permeabilized HL-60 (Human acute promyelocytic leukemia promyeloblast) cells labelling S100A8+S100A9 with Anti-S100A8 + S100A9 antibody [RM1038] ab288715 at 1/500 dilution (0.1ug)(Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor^® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 1/5000 dilution was used as the secondary antibody.

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  Flow Cytometry (Intracellular) - Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
• 

  Immunocytochemistry/ Immunofluorescence - Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)

  Immunofluorescent staining of HeLa cells using anti-AIF RabMAb (Anti-AIF antibody [E20] - Mitochondrial Marker ab32516, left panel) (green) and Rabbit mAb IgG control (ab172730, right panel). DAPI nuclear staining (blue).
  

  Conjugated versions are available for this clone: Alexa Fluor^® 488 (Alexa Fluor® 488 Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab199091), Alexa Fluor^® 647 (Alexa Fluor® 647 Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab199093), R-PE (PE Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab209478), APC (APC Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab232814).

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  Flow Cytometry (Intracellular) - Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
• 

  Immunocytochemistry/ Immunofluorescence - Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)

  Immunocytochemistry/immunofluorescence analysis of HeLa cells with purified Rabbit IgG ab172730 at 1/100. Cells were fixed with 4% paraformaldehyde. An Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/200) was used as the secondary antibody. Counterstained with DAPI (blue).

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  Immunocytochemistry/ Immunofluorescence - Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)

  Immunocytochemistry/immunofluorescence analysis of HeLa cells with unpurified Rabbit IgG ab172730 at 1/10. Cells were fixed with 4% paraformaldehyde. An Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/200) was used as the secondary antibody. Counterstained with DAPI (blue).

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  Flow Cytometry - Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)

  Flow Cytometry staining of Human PBMC (human peripheral blood mononuclear cell) using rabbit Anti- antibody

  Flow cytometric analysis of Human PBMC (human peripheral blood mononuclear cell) cells labelling CD72 with Anti-CD72 antibody [EPR29739-578] ab324351 at 1/500 dilution (0.1ug) / Right compared with a Rabbit monoclonal IgG (ab172730) / Left isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody).

  Goat Anti-Rabbit IgG (Alexa Fluor® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 1/5000 dilution was used as the secondary antibody.

  Gated on viable cells.
  
  Cells are co-stained with CD19 conjugated to Alexa Fluor® 647.

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  Flow Cytometry (Intracellular) - Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)

  Flow Cytometry (Intracellular) staining of C6 (rat glial tumor glial cell) using rabbit Anti- antibody

  Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized C6 (rat glial tumor glial cell) labelling HDAC1 with Anti-HDAC1 antibody [RM1004] - Nuclear Loading Control ab281585 at 1/50 (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

• 

  Flow Cytometry (Intracellular) - Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)

  Flow Cytometry (Intracellular) staining of C6 ( rat glial tumor glial cell) using rabbit Anti- antibody

  Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized C6 (rat glial tumor glial cell) cells labelling HDAC1 with Anti-HDAC1 antibody [EPR23847-170] - Nuclear Loading Control ab280198 at 1/500 dilution (0.1µg) followed by a Goat Anti-Rabbit IgG (Alexa Fluor^® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at a 1/5000 dilution (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

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  Flow Cytometry - Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)

  Flow Cytometry staining using rabbit Anti- antibody

  Flow cytometric analysis of KARPAS-299 (human T cell lymphoma cell) (Right) / HepG2 (human hepatocellular carcinoma epithelial cell) (Left) cells labelling HLA DR + DP + DQ with Anti-HLA DR + DP + DQ antibody [CR3/43] ab7856 at 1/1000 dilution (0.1&micor;g)/Red followed by a secondary antibody Goat Anti-Rabbit IgG (Alexa Fluor^® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) used at a 1/5000 dilution compared with isotype control Rabbit monoclonal IgG (ab172730) / Black and unlabelled control Cell without incubation with primary antibody and secondary antibody / Blue.

  Gated on viable cells.
  Negative control: HepG2.

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  Flow Cytometry (Intracellular) - Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)

  Flow Cytometry (Intracellular) staining using rabbit Anti- antibody

  Flow cytometric analysis of 4% paraformaldehyde fixed, 90% methanol permeabilized SIRT1 KO A549(Human SIRT1 knock out human lung carcinoma epithelial cell, Green) / A549(Megenta) cells labelling SIRT1 with Anti-SIRT1 antibody [19A7AB4] ab110304 at 1/1000 dilution (0.1µg)/Red followed by a Goat Anti-Rabbit IgG (Alexa Fluor^® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at a 1/5000 dilution (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black and Grey) isotype control.

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  Immunoprecipitation - Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)

  KLF5 was immunoprecipitated from 0.35 mg HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate with Anti-KLF5 antibody [EPR29066-77] ab324105 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using Anti-KLF5 antibody [EPR29066-77] ab324105 at 1/1000 dilution.

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

  To minimize protein degradation, cells were lysed immediately after harvest and then applied for Immunoprecipitation as soon as possible.

  All lanes: Immunoprecipitation - Anti-KLF5 antibody [EPR29066-77] (Anti-KLF5 antibody [EPR29066-77] ab324105) at 1/1000 dilution

  Lane 1: HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 10 µg

  Lane 2: Anti-KLF5 antibody [EPR29066-77] ab324105 at 1/30 IP in HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate

  Lane 3: Rabbit monoclonal IgG (ab172730) instead of Anti-KLF5 antibody [EPR29066-77] ab324105 in HeLa whole cell lysate

  Secondary

  All lanes: Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) at 1/5000 dilution

  Observed band size: 50 kDa

  Exposure time: 58s

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  Immunoprecipitation - Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)

  NRAS was immunoprecipitated from 0.35 mg MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate 10 ug with Anti-NRAS antibody [EPR25186-24] ab300431 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using Anti-NRAS antibody [EPR25186-24] ab300431 at 1/1000 dilution.
  

  VeriBlot for IP secondary antibody(HRP)(VeriBlot for IP Detection Reagent (HRP) ab131366) was used at 1/5000 dilution.

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

  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.

  All lanes: Immunoprecipitation - Anti-NRAS antibody [EPR25186-24] (Anti-NRAS antibody [EPR25186-24] ab300431) at 1/1000 dilution

  Lane 1: MCF7 whole cell lysate

  Lane 2: Anti-NRAS antibody [EPR25186-24] ab300431 at 1/30 IP in MCF7 whole cell lysate at 10 µg

  Lane 3: Rabbit monoclonal IgG (ab172730) instead of Anti-NRAS antibody [EPR25186-24] ab300431 in MCF7 whole cell lysate at 10 µg

  Secondary

  All lanes: Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) at 1/5000 dilution

  Developed using the ECL technique.

  Observed band size: 21 kDa

  Exposure time: 180s

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  Flow Cytometry (Intracellular) - Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)

  Flow Cytometry (Intracellular) staining of PC-12 (rat adrenal gland pheochromocytoma cell) using rabbit Anti- antibody

  Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized PC-12 (rat adrenal gland pheochromocytoma cell) cells labelling Histone H3 with Anti-Histone H3 antibody [RM1288] - Nuclear Marker ab322707 at 1/500 dilution (0.1ug) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody).

  Goat Anti-Rabbit IgG (Alexa Fluor® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 1/5000 dilution was used as the secondary antibody.

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  Flow Cytometry (Intracellular) - Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)

  Flow Cytometry (Intracellular) staining of NIH/3T3 (mouse embryonic fibroblast) using rabbit Anti- antibody

  Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized NIH/3T3 (mouse embryonic fibroblast) cells labelling Histone H3 with Anti-Histone H3 antibody [RM1288] - Nuclear Marker ab322707 at 1/500 dilution (0.1ug) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody).

  Goat Anti-Rabbit IgG (Alexa Fluor® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 1/5000 dilution was used as the secondary antibody.

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  Immunoprecipitation - Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)

  Gamma-aminobutyric acid receptor subunit delta was immunoprecipitated from 0.35 mg Mouse cerebellum tissue lysate 10 ug with Anti-GABRD antibody [EPR25324-253] ab300348 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using Anti-GABRD antibody [EPR25324-253] ab300348 at 1/1000 dilution.
  

  VeriBlot for IP secondary antibody(HRP)(VeriBlot for IP Detection Reagent (HRP) ab131366) was used at 1/5000 dilution.

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

  All lanes: Immunoprecipitation - Anti-GABRD antibody [EPR25324-253] (Anti-GABRD antibody [EPR25324-253] ab300348) at 1/1000 dilution

  Lane 1: Mouse cerebellum tissue lysate

  Lane 2: Anti-GABRD antibody [EPR25324-253] ab300348 at 1/30 IP in mouse cerebellum tissue lysate

  Lane 3: Rabbit monoclonal IgG (ab172730) instead of Anti-GABRD antibody [EPR25324-253] ab300348 in mouse cerebellum tissue lysate

  Secondary

  All lanes: Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) at 1/5000 dilution

  Observed band size: 62 kDa

  Exposure time: 3min

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  Flow Cytometry (Intracellular) - Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)

  Flow Cytometry (Intracellular) staining using rabbit Anti- antibody

  Intracellular Flow Cytometry analysis of LNCaP (Human prostate carcinoma epithelial cell) cells labeling PSMA with Anti-PSMA antibody [EP3253] - BSA and Azide free ab247436 at 1/100 dilution (1 µg) (Red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. Goat anti-rabbit IgG (Alexa Fluor® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody was used at 1/2000 dilution. Isotype control - Rabbit monoclonal IgG (ab172730) (Black). Unlabeled control - cell without incubation with primary antibody and secondary antibody (Blue).

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  Immunoprecipitation - Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)

  Gamma-aminobutyric acid receptor subunit delta was immunoprecipitated from 0.35 mg Rat cerebellum tissue lysate 10 ug with Anti-GABRD antibody [EPR25324-253] ab300348 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using Anti-GABRD antibody [EPR25324-253] ab300348 at 1/1000 dilution.
  

  VeriBlot for IP secondary antibody(HRP)(VeriBlot for IP Detection Reagent (HRP) ab131366) was used at 1/5000 dilution.

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

  All lanes: Immunoprecipitation - Anti-GABRD antibody [EPR25324-253] (Anti-GABRD antibody [EPR25324-253] ab300348) at 1/1000 dilution

  Lane 1: Rat cerebellum tissue lysate at 10 µg

  Lane 2: Anti-GABRD antibody [EPR25324-253] ab300348 at 1/30 IP in rat cerebellum tissue lysate at 10 µg

  Lane 3: Rabbit monoclonal IgG (ab172730) instead of Anti-GABRD antibody [EPR25324-253] ab300348 in rat cerebellum tissue lysate at 10 µg

  Secondary

  All lanes: Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) at 1/5000 dilution

  Observed band size: 62 Da

  Exposure time: 41s

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  Immunoprecipitation - Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)

  ASIC1 was immunoprecipitated from 0.35 mg Mouse cerebellum tissue lysate 10 ug with Anti-ASIC1 antibody [EPR25411-161] ab300563 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using Anti-ASIC1 antibody [EPR25411-161] ab300563 at 1/1000 dilution.
  

  VeriBlot for IP secondary antibody(HRP)(VeriBlot for IP Detection Reagent (HRP) ab131366) was used at 1/5000 dilution.

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

  All lanes: Immunoprecipitation - Anti-ASIC1 antibody [EPR25411-161] (Anti-ASIC1 antibody [EPR25411-161] ab300563) at 1/1000 dilution

  Lane 1: Mouse cerebellum tissue lysate at 10 µg

  Lane 2: Anti-ASIC1 antibody [EPR25411-161] ab300563 at 1/30 IP in Mouse cerebellum tissue lysate at 10 µg

  Lane 3: Rabbit monoclonal IgG (ab172730) instead of Anti-ASIC1 antibody [EPR25411-161] ab300563 in mouse cerebellum tissue lysate at 10 µg

  Secondary

  All lanes: Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) at 1/5000 dilution

  Observed band size: 70 kDa, 150 kDa

  Exposure time: 3min

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  Flow Cytometry (Intracellular) - Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)

  Flow Cytometry (Intracellular) staining using rabbit Anti- antibody

  Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized JAR (human placenta choriocarcinoma epithelial cell) treated with 100uM Forskolin for 72 hours (Green) / Untreated control (Magenta) cells labelling hCG beta (pan CGB) with ab324382 at 1/50 dilution (1ug) (Magenta) and Green compared with a Rabbit monoclonal IgG (ab172730) (Black) and Grey isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody).

  Goat Anti-Rabbit IgG (Alexa Fluor® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 1/5000 dilution was used as the secondary antibody.

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  Flow Cytometry (Intracellular) - Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)

  Flow Cytometry (Intracellular) staining using rabbit Anti- antibody

  Intracellular Flow Cytometry analysis of 293T (human embryonic kidney epithelial cell) transfected with myc-tagged PADI4 construct, then treated with 10mM CaCl2 and 10uM Ionomycin for 4h labeling Histone H3 (citrulline R2 + R8 + R17) with Anti-Histone H3 (citrulline R2 + R8 + R17) antibody [RM1001] ab281584 at 1/500 dilution. Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. Goat anti rabbit IgG (Alexa Fluor® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) at 1/2000 dilution was used as a secondary antibody.
  Isotype control : Rabbit monoclonal IgG (ab172730)/left.