Goat Anti-Rabbit IgG H&L (HRP)

38 product images

• 

  Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051)

  Blocking buffer: 5% NFDM/TBST.

  All lanes: Western blot - Anti-RAB29 (phospho T71) antibody [MJF-R24-17-1] (Anti-RAB29 (phospho T71) antibody [MJF-R24-17-1] ab241062) at 1/5000 dilution

  Lane 1: HEK-293 (human embryonic kidney) over-expressing HA-tagged RAB29 and LRRK2 [Y1699C], whole cell lysate at 15 µg

  Lane 2: MLi-2 (100nM for 60min) treated HEK-293 over-expressing HA-tagged RAB29 and LRRK2 [Y1699C], whole cell lysate at 15 µg

  Secondary

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

  Predicted band size: 23 kDa

  Observed band size: 23 kDa

• 

  Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051)

  The basal expression level of AKT1 (phospho S473) varies in different cell lines reported by PMID: 19372546. But to detect clear signal, treatment is strongly recommended when using this antibody.
  

  Blocking and Diluting buffer: 5% NFDM/TBST

  All lanes: Western blot - Anti-AKT1 (pS473) + AKT2 (pS474) + AKT3 (pS472) antibody [EPR18853] (Anti-AKT1 (pS473) + AKT2 (pS474) + AKT3 (pS472) antibody [EPR18853] ab192623) at 1/1000 dilution

  Lane 1: LNCaP (Human prostate carcinoma epithelial cell) whole cell lysates at 20 µg

  Lane 2: LNCaP (Human prostate carcinoma epithelial cell) treated with 0.1 uM Calyculin A for 30 minutes whole cell lysates at 20 µg

  Lane 3: A549 (Human lung carcinoma epithelial cell) whole cell lysates at 20 µg

  Lane 4: MCF7 (Human breast adenocarcinoma epithelial cell) whole cell lysates at 20 µg

  Lane 5: HepG2 (Human hepatocellular carcinoma epithelial cell) whole cell lysates at 20 µg

  Lane 6: HUVEC (Human umbilical vein endothelial cell) whole cell lysates at 20 µg

  Lane 7: C2C12 (Mouse myoblasts myoblast) whole cell lysates at 20 µg

  Lane 8: Mouse brain lysates at 20 µg

  Lane 9: Rat heart lysates at 20 µg

  Secondary

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

  Predicted band size: 56 kDa

  Observed band size: 56 kDa

  Exposure time: 50s

• 

  Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051)

  This data was developed using Anti-ALDH3A1 antibody [EPR7405] ab128902, the same antibody clone in a different buffer formulation.

  All lanes: Western blot - Anti-ALDH3A1 antibody [EPR7405] (Anti-ALDH3A1 antibody [EPR7405] ab128902) at 1/5000 dilution

  All lanes: A549 (Human lung carcinoma epithelial cell) whole cell lysate at 15 µg

  Secondary

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

  Predicted band size: 50 kDa

  Observed band size: 50 kDa

• 

  Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051)

  All lanes: Western blot - Anti-Nucleolin (phospho T84) antibody [EPR8080] (Anti-Nucleolin (phospho T84) antibody [EPR8080] ab155977) at 1/5000 dilution

  Lane 1: Untreated K-562 (Human chronic myelogenous leukemia lymphoblast) whole cell lysate at 15 µg

  Lane 2: K-562 (Human chronic myelogenous leukemia lymphoblast) treated with 100nM Calyculin A for 30 minutes whole cell lysate at 15 µg

  Lane 3: K-562 (Human chronic myelogenous leukemia lymphoblast) treated with 100nM Calyculin A for 30 minutes, then the membrane treated with Lambda Phosphatase for 1 hour at 15 µg

  Secondary

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

  Predicted band size: 76 kDa

• 

  Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051)

  This data was developed using Anti-CXCL7/PBP antibody [EPR19780] ab206966, the same antibody clone in a different buffer formulation.

  All lanes: Western blot - Anti-CXCL7/PBP antibody [EPR19780] (Anti-CXCL7/PBP antibody [EPR19780] ab206966) at 1/1000 dilution

  All lanes: Human NAP2 active recombinant protein at 0.01 µg

  Secondary

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

  Predicted band size: 14 kDa

  Observed band size: 11 kDa

  Exposure time: 5s

• 

  Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051)

  Blocking/Dilution buffer: 5% NFDM/TBST.

  Active human PKC alpha full length protein (Catalog#Recombinant human PKC alpha protein (Active) ab55672) contains aa1-672 with GST-tag; Active human PKC gamma full length protein (Catalog#Recombinant human PKC gamma protein ab60842) contains aa1-677 with GST-tag.

  All lanes: Western blot - Anti-PKC alpha + beta 2 + gamma antibody [EPR18104] (Anti-PKC alpha + beta 2 + gamma antibody [EPR18104] ab184746) at 1/1000 dilution

  Lane 1: Active human PKC alpha full length protein at 0.02 µg

  Lane 2: Active human PKC gamma full length protein at 0.02 µg

  Secondary

  All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution

  Predicted band size: 77 kDa, 78 kDa

  Exposure time: 1s

• 

  Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051)

  This data was developed using Anti-CASZ1 antibody [EPR23824-4] ab259847, the same antibody clone in a different buffer formulation.
  

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

  Negative control: Mouse liver (PMID: 16631614)

  The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID: 33594190, 32060262)

  Exposure time: 81 seconds

  All lanes: Western blot - Anti-CASZ1 antibody [EPR23824-4] (Anti-CASZ1 antibody [EPR23824-4] ab259847) at 1/1000 dilution

  Lane 1: Mouse lung tissue lysate at 20 µg

  Lane 2: Mouse E18 heart tissue lysate at 20 µg

  Lane 3: Mouse P20 retina tissue lysate at 20 µg

  Lane 4: Mouse liver tissue lysate at 20 µg

  Secondary

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

  Predicted band size: 190 kDa

  Observed band size: 150 kDa, 250 kDa

• 

  Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051)

  This data was developed using Anti-PKA alpha/beta/gamma (catalytic subunit) (phospho T197) antibody [EP2606Y] ab75991, the same antibody clone in a different buffer formulation.

  All lanes: Western blot - Anti-PKA alpha/beta/gamma (catalytic subunit) (phospho T197) antibody [EP2606Y] (Anti-PKA alpha/beta/gamma (catalytic subunit) (phospho T197) antibody [EP2606Y] ab75991) at 1/1000 dilution

  Lane 1: Untreated HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate at 15 µg

  Lane 2: HeLa (Human cervix adenocarcinoma epithelial cell) starved overnight, then treated with 100nM Calyculin A for 30 minutes, whole cell lysate at 15 µg

  Lane 3: HeLa (Human cervix adenocarcinoma epithelial cell) starved overnight, then treated with 100nM Calyculin A for 30 minutes whole cell lysate, then the membrane treated with Lambda Phosphatase for 1 hour at 15 µg

  Secondary

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

  Predicted band size: 40 kDa

  Observed band size: 40 kDa

• 

  Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051)

  All lanes: Western blot - Anti-Cytochrome P450 1A2 antibody [EPR6138(2)] (Anti-Cytochrome P450 1A2 antibody [EPR6138(2)] ab151728) at 1/2000 dilution

  All lanes: HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates at 15 µg

  Secondary

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

  Predicted band size: 58 kDa

  Observed band size: 58 kDa

• 

  Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051)

  This data was developed using Anti-alpha 1 Fetoprotein antibody [EPR26548-19] ab290637, the same antibody clone in different buffer formulation.
  

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

  All lanes: Western blot - Anti-alpha 1 Fetoprotein antibody [EPR26548-19] (Anti-alpha 1 Fetoprotein antibody [EPR26548-19] ab290637) at 1000 Cells

  Lane 1: Untreated Hepa1-6 (mouse hepatoma epithelial cell) whole cell lysate at 20 µg

  Lane 2: Hepa1-6 treated with 300ng/ml Brefeldin A for 24h whole cell lysate at 20 µg

  Secondary

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

  Predicted band size: 68 kDa

  Observed band size: 70 kDa

  Exposure time: 37s

• 

  Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051)

  This data was developed using Anti-PU.1/Spi1 antibody [EPR3158Y] ab76543, the same antibody clone in a different buffer formulation.
  

  PU.1 is a 37-42 kDa member of the ets family of transcription factors. The molecule weight observed is consistent with what has been described in the literature (PMID: 28362429, 31586032, 28586009).

  All lanes: Western blot - Anti-PU.1/Spi1 antibody [EPR3158Y] (Anti-PU.1/Spi1 antibody [EPR3158Y] ab76543) at 1/1000 dilution

  Lane 1: Daudi (Human Burkitt's lymphoma lymphoblast) whole cell lysate at 20 µg

  Lane 2: THP-1 (Human monocytic leukemia monocyte) whole cell lysate at 20 µg

  Lane 3: Raji (Human Burkitt's lymphoma B lymphocyte) whole cell lysate at 20 µg

  Secondary

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

  Predicted band size: 31 kDa

  Observed band size: 37-42 kDa

• 

  Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051)

  This data was produced using Anti-LYVE1 antibody [RM1008] ab281587, the same clone but in a different formulation.
  

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

  Several bands are observed including soluble, glycosylated and non-glycosylated forms which are consistent with the literature (PMID: 26966180).

  Exposure time: 37 seconds

  Lanes 1 and 3: Mouse lung lysate at 20 µg

  Lane 2: Mouse brain lysate at 20 µg

  Secondary

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

• 

  Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051)

  Exposure times: Lanes 1,2: 1 second; Lanes 3-6: 5 seconds.

  Blocking/Dilution buffer: 5% NFDM/TBST.

  All lanes: Western blot - Anti-RPS6 (phospho S240 + S244) antibody [EPR20770] (Anti-RPS6 (phospho S240 + S244) antibody [EPR20770] ab215214) at 1/5000 dilution

  Lane 1: MCF7 (human breast adenocarcinoma cell line) grown in serum-free media overnight, whole cell lysate at 20 µg

  Lane 2: MCF7 grown in serum-free media overnight, then grown in 20% FBS media for 30 minutes, whole cell lysate at 20 µg

  Lane 3: NIH/3T3 (mouse embryonic fibroblast cell line) grown in serum-free media for 4 hours, whole cell lysate at 20 µg

  Lane 4: NIH/3T3 grown in serum-free media for 4 hours, then grown in 20% FBS media for 30 minutes, whole cell lysate at 20 µg

  Lane 5: PC-12 (rat adrenal gland pheochromocytoma cell line) grown in serum-free media overnight, whole cell lysate at 20 µg

  Lane 6: PC-12 grown in serum-free media overnight, then grown in 20% FBS media for 30 minutes, whole cell lysate at 20 µg

  Secondary

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

  Developed using the ECL technique.

  Predicted band size: 29 kDa

  Observed band size: 32 kDa

• 

  Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051)

  This data was produced using Anti-Sodium Potassium ATPase antibody [EP1845Y] - Plasma Membrane Loading Control ab76020, the same antibody clone in a different formulation
  

  Blocking/Diluting buffer and concentration 5% NFDM/TBST

  We suggest not to boil the sample after lysis.

  All lanes: Western blot - Anti-Sodium Potassium ATPase antibody [EP1845Y] - BSA and Azide free (Anti-Sodium Potassium ATPase antibody [EP1845Y] - BSA and Azide free ab167390) at 1/100000 dilution

  Lane 1: HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate prepared from RIPA lysis method at 15 µg

  Lane 2: HeLa whole cell lysate prepared from 1% SDS HOT lysis method at 15 µg

  Lane 3: HeLa whole cell lysate prepared from RIPA lysis method at 15 µg

  Lane 4: HeLa whole cell lysate prepared from 1%SDS HOT lysis method at 15 µg

  Lane 5: Raw264.7 (Mouse abelson murine leukemia virus-induced tumor) whole cell lysate prepared from RIPA lysis method at 15 µg

  Lanes 6 and 8: Raw264.7 whole cell lysate prepared from 1%SDS HOT lysis method at 15 µg

  Lane 7: Raw264.7 whole cell lysate prepared from RIPA lysis method at 15 µg

  Secondary

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

  Predicted band size: 113 kDa

  Observed band size: 100 kDa

  Exposure time: 10s

• 

  Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051)

  Exposure times: Lane 1: 1 minute; Lane 2: 20 seconds; Lane 3: 10 seconds.

  Blocking/Dilution buffer: 5% NFDM/TBST.

  The blot was developed on a BIO-RAD® ChemiDoc™ MP instrument.

  All lanes: Western blot - Anti-Thrombomodulin antibody [EPR18217-209] (Anti-Thrombomodulin antibody [EPR18217-209] ab230010) at 1/1000 dilution

  Lane 1: Mouse lung tissue lysate at 20 µg

  Lane 2: bEND.3 (mouse brain endothelioma cell line) whole cell lysate at 10 µg

  Lane 3: Mouse placenta tissue lysate at 10 µg

  Secondary

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

  Lanes 2 - 3: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution

  Developed using the ECL technique.

  Predicted band size: 60 kDa

  Observed band size: 105 kDa, 75 kDa

• 

  Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051)

  Blocking/Dilution buffer: 5% NFDM/TBST.

  All lanes: Western blot - Anti-CRISPR-Cas9 antibody [EPR19633] (Anti-CRISPR-Cas9 antibody [EPR19633] ab202657) at 1/20000 dilution

  Lane 1: Empty vector with GFP-Myc tag (vector control) transfected HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate at 20 µg

  Lane 2: HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate transfected with CRISPR-Cas9 (Q99ZW2, Streptococcus pyogenes serotype M1) with GFP-Myc tag at 20 µg

  Lane 3: HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate transfected with CRISPR-Cas9 (G3ECR1, Streptococcus thermophilus, N–terminal aa1-800) with GFP-Myc tag at 20 µg

  Lane 4: HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate transfected with CRISPR-Cas9 (G3ECR1, Streptococcus thermophilus, C–terminal aa801-1409) with GFP-Myc tag at 20 µg

  Lane 5: Empty vector with Myc-His tag (vector control) transfected HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate at 20 µg

  Lane 6: HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate transfected with CRISPR-Cas9 (A1IQ68, Neisseria meningitidis serogroup A / serotype 4A (strain Z2491)) with Myc-His tag at 20 µg

  Lane 7: HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate transfected with CRISPR-Cas9 (G3ECR1, Streptococcus thermophilus) with Myc-His tag at 20 µg

  Lane 8: HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate transfected with CRISPR-Cas9 (Q03JI6, Streptococcus thermophilus (strain ATCC BAA-491 / LMD-9)) with Myc-His tag at 20 µg

  Lane 9: HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate transfected with CRISPR-Cas9 (J7RUA5, Staphylococcus aureus subsp. aureus) with Myc-His tag at 20 µg

  Secondary

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

  Predicted band size: 164 kDa

  Exposure time: 5s

• 

  Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051)

  All lanes: Western blot - Anti-cAMP Protein Kinase Catalytic subunit antibody [EP2102Y] (Anti-cAMP Protein Kinase Catalytic subunit antibody [EP2102Y] ab76238) at 1/20000 dilution

  Lane 1: MCF7 (Human breast adenocarcinoma epithelial cell) whole cell lysates at 15 µg

  Lane 2: NIH/3T3 (Mouse embryonic fibroblast) whole cell lysates at 15 µg

  Lane 3: C6 (Rat glial tumor glial cell) whole cell lysates at 15 µg

  Secondary

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

  Predicted band size: 41 kDa

  Observed band size: 42 kDa

• 

  Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051)

  This data was developed using Anti-VIRMA/KIAA1429 antibody [EPR24279-23] ab271136, the same antibody clone in a different buffer formulation.
  

  The observed MW are consistent with what has been described in the literature (PMID: 34490238).

  Blocking and dilution buffer: 5% NFDM/TBST.

  All lanes: Western blot - Anti-VIRMA/KIAA1429 antibody [EPR24279-23] (Anti-VIRMA/KIAA1429 antibody [EPR24279-23] ab271136) at 1/1000 dilution

  Lane 1: PC-3 (human prostate adenocarcinoma epithelial cell), whole cell lysate at 20 µg

  Lane 2: SK-OV-3 (human ovarian cancer epithelial cell), whole cell lysate at 20 µg

  Lane 3: T-47D (human ductal breast epithelial tumor epithelial cell), whole cell lysate at 20 µg

  Lane 4: C2C12 (mouse myoblasts), whole cell lysate at 20 µg

  Lane 5: Mouse liver cancer lysate at 20 µg

  Secondary

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

  Predicted band size: 202 kDa

  Observed band size: 202 kDa

  Exposure time: 26s

• 

  Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051)

  This data was developed using Anti-SMAD1 + SMAD5 + SMAD9 (phospho S463 + S465 + S467) antibody [EP728(2)AY] ab76296, the same antibody clone in a different buffer formulation.

  All lanes: Western blot - Anti-SMAD1 + SMAD5 + SMAD9 (phospho S463 + S465 + S467) antibody [EP728(2)AY] (Anti-SMAD1 + SMAD5 + SMAD9 (phospho S463 + S465 + S467) antibody [EP728(2)AY] ab76296) at 1/1000 dilution

  Lane 1: Untreated C6 (Rat glial tumor glial cell) whole cell lysate at 15 µg

  Lane 2: C6 treated with 10ng/ml TGF beta 3 and 50µM MG-132 for 30 min whole cell lysate at 15 µg

  Lane 3: C6 treated with 10ng/ml TGF beta 3 and 50µM MG-132 for 30 min whole cell lysate, then the membrane treated with Alkaline Phosphatase for 1 hour at 15 µg

  Secondary

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

  Observed band size: 52 kDa

  Exposure time: 7s

• 

  Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051)

  This blot was produced using a 10% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 5% Bovine Serum Albumin before being incubated with Anti-CYP1A1 antibody ab124295 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution.

  All lanes: Western blot - Anti-CYP1A1 antibody (Anti-CYP1A1 antibody ab124295) at 1 µg/mL

  Lanes 1 and 3: Lung (Human) Tissue Lysate at 10 µg

  Lanes 2 and 4: Human placenta tissue lysate - total protein (ab29745) at 10 µg

  Secondary

  All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution

  Developed using the ECL technique.

  Performed under reducing conditions.

  Predicted band size: 58 kDa

  Observed band size: 48 kDa, 55 kDa, 73 kDa

  Exposure time: 90s

• 

  Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051)

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

  Anti-CD130 (gp130) antibody [EPR24557-50] ab283685 was shown to specifically react with IL6ST (CD130/gp130) in wild-type A549 cells. Loss of signal was observed when knockout cell line Human IL6ST (CD130/gp130) knockout A549 cell line ab266938 (knockout cell lysate Human IL6ST (CD130/gp130) knockout A549 cell lysate ab257207) was used. Wild-type and IL6ST (CD130/gp130) knockout samples were subjected to SDS-PAGE. Anti-CD130 (gp130) antibody [EPR24557-50] ab283685 and Anti-GAPDH antibody [EPR16891] - Loading Control ab181602 (Rabbit anti-GAPDH loading control) were incubated for 1 hour at room temperature at 1/1000 dilution and 1/200, 000 dilution respectively. Blots were developed with Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) secondary antibody at 1/50, 000 dilution and 1/100, 000 dilution for 1 hour at room temperature before imaging. The blot was developed on a BIO-RAD® ChemiDoc™ MP instrument using the ECL technique.

  Samples are non-boiled as boiling may cause protein aggregates.

  Exposure time: 70 seconds

  All lanes: Western blot - Anti-CD130 (gp130) antibody [EPR24557-50] (Anti-CD130 (gp130) antibody [EPR24557-50] ab283685) at 1/1000 dilution

  Lane 1: Wild-type A549 (human lung carcinoma epithelial cell) whole cell lysate at 20 µg

  Lane 2: IL6ST (CD130/gp130) knockout A549 (human lung carcinoma epithelial cell) whole cell lysate at 20 µg

  Lane 3: PC-3 (human prostate adenocarcinoma epithelial cell) whole cell lysate at 20 µg

  Secondary

  All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution

  Predicted band size: 104 kDa

  Observed band size: 130 kDa

• 

  Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051)

  Blocking/Dilution buffer: 5% NFDM/TBST.

  All lanes: Western blot - Anti-RNA polymerase beta antibody [EPR18704] (Anti-RNA polymerase beta antibody [EPR18704] ab191598) at 1/2000 dilution

  All lanes: E.coli whole cell lysates at 10 µg

  Secondary

  All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution

  Predicted band size: 150 kDa

  Observed band size: 150 kDa

  Exposure time: 30s

• 

  Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051)

  Active human PKC alpha full length protein (Catalog#Recombinant human PKC alpha protein (Active) ab55672) contains aa1-672 with GST-tag; Active human PKC beta 1 full length protein(Catalog#Recombinant human PKC beta 1 protein ab60840) contains aa1-671 with GST-tag; Active human PKC beta 2 full length protein(Catalog#Recombinant human PKC beta 2 protein ab60841) contains aa1-673 with GST-tag; Active human PKC gamma full length protein(Catalog#Recombinant human PKC gamma protein ab60842) contains aa1-677 with GST-tag; Active human PKC delta full length protein(Catalog#Recombinant human PKC delta protein ab60844) contains aa1-676 with GST-tag; Active human PKC eta full length protein(Catalog#Recombinant human PKC eta protein ab60849) contains aa1-683 with GST-tag; Active human PKC epsilon full length protein(Catalog#Recombinant human PKC epsilon protein ab60847) contains aa1-737 with GST-tag; Active human PKC theta full length protein(Catalog#Recombinant human PKC theta/PRKCQ protein ab56641) contains aa1-706 with GST-tag; Active human PKC mu full length protein(Catalog#Recombinant human PKC mu/PKD protein ab60873) contains aa1-912 with GST-tag.
  

  Blocking/Diluting buffer 5% NFDM/TBST

  All lanes: Western blot - Anti-PKC antibody [EPR16898] (Anti-PKC antibody [EPR16898] ab179522) at 1/2000 dilution

  Lane 1: Active human PKC alpha full length protein at 0.02 µg

  Lane 2: Active human PKC beta 1 full length protein at 0.02 µg

  Lane 3: Active human PKC beta 2 full length protein at 0.02 µg

  Lane 4: Active human PKC gamma full length protein at 0.02 µg

  Lane 5: Active human PKC delta full length protein at 0.02 µg

  Lane 6: Active human PKC eta full length protein at 0.02 µg

  Lane 7: Active human PKC epsilon full length protein at 0.02 µg

  Lane 8: Active human PKC theta full length protein at 0.02 µg

  Lane 9: Active human PKC mu full length protein at 0.02 µg

  Secondary

  All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution

  Predicted band size: 77 kDa

  Exposure time: 2s

• 

  Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051)

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

  All lanes: Western blot - Anti-Netrin 1 antibody [EPR5428] (Anti-Netrin 1 antibody [EPR5428] ab126729) at 1/500 dilution

  Lane 1: Rat brain lysates prepared in RIPA lysis method at 20 µg

  Lane 2: Rat brain lysates prepared in 1%SDS Hot lysis method at 20 µg

  Secondary

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

  Developed using the ECL technique.

  Predicted band size: 67 kDa

  Observed band size: 75 kDa

  Exposure time: 180s

• 

  Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051)

  All lanes: Western blot - Anti-DNA Polymerase beta antibody [EPR12722(B)] (Anti-DNA Polymerase beta antibody [EPR12722(B)] ab175197) at 1/1000 dilution

  Lane 1: HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate at 20 µg

  Lane 2: Jurkat (Human T cell leukemia T lymphocyte) whole cell lysate at 20 µg

  Lane 3: K-562 (Human chronic myelogenous leukemia lymphoblast) whole cell lysate at 20 µg

  Lane 4: A431 (Human epidermoid carcinoma epithelial cell) whole cell lysate at 20 µg

  Lane 5: Mouse brain lysate at 20 µg

  Lane 6: Mouse spleen lysate at 20 µg

  Lane 7: Rat brain lysate at 20 µg

  Lane 8: Rat spleen lysate

  Lane 9: C6 (Rat glial tumor glial cell) whole cell lysate

  Secondary

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

  Predicted band size: 38 kDa

  Observed band size: 38 kDa

• 

  Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051)

  This antibody shows low affinity on HeLa cells and detects a strong non-specific band around 75 KD.

  All lanes: Western blot - Anti-ARF6 antibody [EPR8357] (Anti-ARF6 antibody [EPR8357] ab131261) at 1/1000 dilution

  All lanes: HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates at 15 µg

  Secondary

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

  Predicted band size: 20 kDa

  Observed band size: 18 kDa

• 

  Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051)

  All lanes: Western blot - Anti-DDX17 antibody [EPR13807(B)] (Anti-DDX17 antibody [EPR13807(B)] ab180190) at 1/1000 dilution

  Lane 1: HEK-293 (Human embryonic kidney epithelial cell) whole cell lysate at 20 µg

  Lane 2: MCF7 (Human breast adenocarcinoma epithelial cell) whole cell lysate at 20 µg

  Lane 3: Rat brain lysate at 20 µg

  Lane 4: Rat testis lysate at 20 µg

  Secondary

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

  Predicted band size: 72 kDa

• 

  Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051)

  Blocking buffer: 5% NFDM /TBST

  All lanes: Western blot - Anti-RNF185+RNF5/NG2 antibody [EPR14070] (Anti-RNF185+RNF5/NG2 antibody [EPR14070] ab181227) at 1/1000 dilution

  Lane 1: Human RNF185 full-length ORF recombinant protein with GST-tag at N-terminal at 0.01 µg

  Lane 2: Human RNF5 full-length ORF recombinant protein with GST-tag at N-terminal at 0.01 µg

  Secondary

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

  Developed using the ECL technique.

  Predicted band size: 20 kDa

  Observed band size: 47.45 kDa

  Exposure time: 1s

• 

  Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051)

  Blocking and dilution buffer: 5% NFDM/TBST.
  

  Exposure times:

  Lanes 1-3: 10 seconds.

  Lane 4: 8 seconds.

  All lanes: Western blot - Anti-Histone H4 (mono methyl R92) antibody [EPR21782] (Anti-Histone H4 (mono methyl R92) antibody [EPR21782] ab219797) at 1/1000 dilution

  Lane 1: NIH/3T3 (mouse embryo fibroblast cell line) whole cell lysate at 20 µg

  Lane 2: C6 (rat glioma cell line) whole cell lysate at 20 µg

  Lane 3: PC-12 (rat adrenal gland pheochromocytoma cell line) whole cell lysate at 20 µg

  Lane 4: HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg

  Secondary

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

  Predicted band size: 11 kDa

  Observed band size: 11 kDa

• 

  Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051)

  The bands detected are consistent with what have been described in the literatures (PMID: 11741954, 27033701). Full-length Integrin alpha V: 150kDa; Integrin alpha V light chain: 27-30kDa

  All lanes: Western blot - Anti-Integrin alpha V antibody [EPR5583] (Anti-Integrin alpha V antibody [EPR5583] ab124968) at 1/1000 dilution

  Lane 1: A549 (Human lung carcinoma epithelial cell) whole cell lysate at 20 µg

  Lane 2: A431 (Human epidermoid carcinoma epithelial cell) whole cell lysate at 20 µg

  Lane 3: HUVEC (Human umbilical vein endothelial cell) whole cell lysate at 20 µg

  Lane 4: U-87 MG (Human glioblastoma-astrocytoma epithelial cell) whole cell lysate at 20 µg

  Lane 5: NIH/3T3 (Mouse embryonic fibroblast) whole cell lysate at 20 µg

  Lane 6: C6 (Rat glial tumor glial cell) whole cell lysate at 20 µg

  Secondary

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

  Predicted band size: 116 kDa

  Observed band size: 150 kDa, 27-30 kDa

• 

  Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051)

  This data was developed using Anti-EpCAM antibody [RM1016] ab282457, the same antibody clone in a different buffer formulation.
  

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

  Negative control: HeLa (PMID: 31806375), NIH/3T3 (PMID: 23264216).

  The MW observed is consistent with the literature: PMID 19136966; PMID 23618806. The band between 12-15kDa has been documented in literature as the cleaved form of EpCAM (PMID: 23409978).

  Exposure time: 8 seconds.

  All lanes: Western blot - Anti-EpCAM antibody [RM1016] - BSA and Azide free (Anti-EpCAM antibody [RM1016] - BSA and Azide free ab283866) at 1/1000 dilution

  Lane 1: HCT 116 (Human colorectal carcinoma epithelial cell) whole cell lysate at 20 µg

  Lane 2: HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate at 20 µg

  Lane 3: 4T1 (Mouse mammary gland carcinoma epithelial cell) whole cell lysate at 20 µg

  Lane 4: NIH/3T3 (Mouse embryonic fibroblast) whole cell lysate at 20 µg

  Lane 5: Human colon lysate at 20 µg

  Lane 6: Mouse colon lysate at 20 µg

  Lane 7: Rat colon lysate at 20 µg

  Lane 8: Mouse kidney lysate at 20 µg

  Lane 9: Rat kidney lysate at 20 µg

  Secondary

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

  Predicted band size: 35 kDa

  Observed band size: 12 kDa, 36 kDa, 40 kDa

• 

  Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051)

  Blocking/Dilution buffer: 5% NFDM/TBST.
  

  Active human PKC alpha full length protein (Catalog#Recombinant human PKC alpha protein (Active) ab55672) contains aa1-672 with GST-tag; Active human PKC beta 1 full length protein (Catalog#Recombinant human PKC beta 1 protein ab60840) contains aa1-671 with GST-tag; Active human PKC beta 2 full length protein (Catalog#Recombinant human PKC beta 2 protein ab60841) contains aa1-673 with GST-tag; Active human PKC gamma full length protein (Catalog#Recombinant human PKC gamma protein ab60842) contains aa1-677 with GST-tag; Active human PKC delta full length protein (Catalog#Recombinant human PKC delta protein ab60844) contains aa1-676 with GST-tag; Active human PKC mu full length protein (Catalog#Recombinant human PKC mu/PKD protein ab60873) contains aa1-912 with GST-tag.

  All lanes: Western blot - Anti-PKC antibody [EPR16791] (Anti-PKC antibody [EPR16791] ab179523) at 1/2000 dilution

  Lane 1: Active human PKC alpha full length protein at 0.02 µg

  Lane 2: Active human PKC beta 1 full length protein at 0.02 µg

  Lane 3: Active human PKC beta 2 full length protein at 0.02 µg

  Lane 4: Active human PKC gamma full length protein at 0.02 µg

  Lane 5: Active human PKC delta full length protein at 0.02 µg

  Lane 6: Active human PKC mu full length protein at 0.02 µg

  Secondary

  All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution

  Predicted band size: 77 kDa

  Exposure time: 3min

• 

  Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051)

  This data was developed using Anti-beta Catenin non-phospho S37/T41 antibody [EPR23969-131] ab246504, the same antibody clone in a different buffer formulation.
  

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

  Exposure time 92 seconds

  An extra band around 45 kDa was observed.

  All lanes: Western blot - Anti-beta Catenin non-phospho S37/T41 antibody [EPR23969-131] (Anti-beta Catenin non-phospho S37/T41 antibody [EPR23969-131] ab246504) at 1/1000 dilution

  Lane 1: HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate at 20 µg

  Lane 2: HCT 116 (human colorectal carcinoma epithelial cell) whole cell lysate at 20 µg

  Lane 3: LLC1 (mouse lung carcinoma ) whole cell lysate at 20 µg

  Lane 4: C6 (rat glial tumor glial cell) whole cell lysate at 20 µg

  Secondary

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

  Predicted band size: 85 kDa

  Observed band size: 95 kDa

• 

  Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051)

  The bands observed are consistent with what have been described in PMID 30649915 as isoforms of p63.
  

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-p63 antibody [EPR5701] ab124762).

  All lanes: Western blot - Anti-p63 antibody [EPR5701] (Anti-p63 antibody [EPR5701] ab124762) at 1/1000 dilution

  Lane 1: HaCaT (Human skin keratinocyte) whole cell lysates at 20 µg

  Lane 2: A431 (Human epidermoid carcinoma epithelial cell) whole cell lysates at 20 µg

  Lane 3: Mouse skin lysates at 20 µg

  Lane 4: Mouse thymus lysates at 20 µg

  Lane 5: Mouse bladder lysates at 20 µg

  Lane 6: Rat skin lysates at 20 µg

  Lane 7: Rat bladder lysates at 20 µg

  Lane 8: PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysates at 20 µg

  Secondary

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

  Predicted band size: 77 kDa

  Observed band size: 37-75 kDa

• 

  Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051)

  The molecular weight observed is consistent with what has been described in the literature (PMID: 10809957).
  

  Negative control: SH-SY5Y (PMID: 26749374), K562 (PMID: 21037579).

  Blocking/Dilution buffer: 5% NFDM/TBST.

  All lanes: Western blot - Anti-CD1d antibody [EPR22526-9] (Anti-CD1d antibody [EPR22526-9] ab256344) at 1/1000 dilution

  Lane 1: Jurkat (human T cell leukemia T lymphocyte), whole cell lysate at 20 µg

  Lane 2: MOLT-4 (human lymphoblastic leukemia T lymphoblast), whole cell lysate at 20 µg

  Lane 3: SH-SY5Y (human neuroblastoma epithelial cell), whole cell lysate at 20 µg

  Lane 4: K562 (human chronic myelogenous leukemia lymphoblast), whole cell lysate at 20 µg

  Secondary

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

  Predicted band size: 38 kDa

  Observed band size: 48 kDa

  Exposure time: 37s

• 

  Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051)

  This data was developed using Anti-Nav1.5/SCN5A antibody [EPR25136-48] ab300048, the same antibody clone in a different buffer formulation.
  

  5% NFDM/TBST was used as a blocking and diluting buffer and concentration.

  Negative control: liver, spleen(PMID: 1309946)
  
  Samples are non-boiled as boiling may cause protein aggregates.
  
  This blot was developed using a higher sensitivity ECL substrate.

  All lanes: Western blot - Anti-Nav1.5/SCN5A antibody [EPR25136-48] (Anti-Nav1.5/SCN5A antibody [EPR25136-48] ab300048) at 1/1000 dilution

  Lane 1: Mouse heart tissue lysate at 20 µg

  Lane 2: Mouse liver tissue lysate at 20 µg

  Lane 3: Mouse spleen tissue lysate at 20 µg

  Secondary

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

  Predicted band size: 226 kDa

  Observed band size: 250 kDa

  Exposure time: 3min

• 

  Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051)

  This data was developed using 259919, the same antibody clone in a different buffer formulation.
  

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

  Negative control: HL-60 (PMID: 10445843 )

  The two reactive bands represent different isoforms

  Exposure time: 37 seconds.

  All lanes: Western blot - Anti-L3MBTL1 antibody [EPR23810-121] (Anti-L3MBTL1 antibody [EPR23810-121] ab259919) at 1/1000 dilution

  Lane 1: IMR-32 (human neuroblastoma neuroblast) whole cell lysate at 20 µg

  Lane 2: SH-SY5Y (human neuroblastoma epithelial cell) whole cell lysate at 20 µg

  Lane 3: SW480 (human colorectal adenocarcinoma epithelial cell) whole cell lysate at 20 µg

  Lane 4: HL-60 (human Acute Promyelocytic Leukemia promyeloblast) whole cell lysate at 20 µg

  Lane 5: Neuro-2a (mouse neuroblastoma neuroblast) whole cell lysate at 20 µg

  Lane 6: NIH/3T3 (mouse embryonic fibroblast) whole cell lysate at 20 µg

  Lane 7: C6 (rat glial tumor glial cell) whole cell lysate at 20 µg

  Secondary

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

  Predicted band size: 83 kDa

  Observed band size: 100 kDa

• 

  Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051)

  Blocking/Dilution buffer: 5% NFDM/TBST.

  All lanes: Western blot - Anti-SF3B3 antibody [EPR18441] (Anti-SF3B3 antibody [EPR18441] ab209403) at 1/5000 dilution

  Lane 1: HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg

  Lane 2: Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate at 20 µg

  Secondary

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

  Predicted band size: 136 kDa

  Observed band size: 136 kDa

  Exposure time: 8s