Anti-GAPDH antibody [EPR16891] - Loading Control

37 product images

• 

  Immunoprecipitation - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)

  GAPDH was immunoprecipitated from 1mg of HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell extract with ab181602 at 1/60 dilution. Western blot was performed from the immunoprecipitate using ab181602 at 1/5000 dilution. Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG, was used as secondary antibody at 1/1500 dilution. Lane 1: HeLa whole cell extract.
  

  Lane 2: PBS instead of HeLa whole cell extract.

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

  All lanes: Immunoprecipitation - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)

  Predicted band size: 36 kDa

  Observed band size: 36 kDa

• 

  Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)

  Blocking/Dilution buffer: 5% NFDM/TBST.

  All lanes: Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) at 1/10000 dilution

  Lane 1: Mouse kidney lysates at 10 µg

  Lane 2: Mouse spleen lysates at 10 µg

  Lane 3: RAW 264.7 (Mouse macrophage cells transformed with Abelson murine leukemia virus) whole cell lysates at 10 µg

  Lane 4: PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysates at 10 µg

  Lane 5: Rat brain lysates at 10 µg

  Secondary

  All lanes: Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

  Predicted band size: 36 kDa

  Observed band size: 36 kDa

• 

  Immunocytochemistry/ Immunofluorescence - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)

  Immunocytochemistry/immunofluorescence staining of 4% paraformaldehyde fixed; 0.1% triton X 100 permeabilized HeLa (human cervix adenocarcinoma) cells labeling GAPDH with ab181602 at dilution of 1/500. The secondary antibody used was Alexa Fluor^® 488; goat anti-rabbit IgG (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) at a dilution of 1/400. Nucleus was counter-stained with DAPI (blue). Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291, a mouse anti-tubulin antibody (1/500) was used to stain tubulin along with Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120 (AlexaFluor^®594 goat anti-mouse secondary, 1/500). The negative controls are shown in the bottom middle and right hand panels- for negative control 1 primary antibody (ab181602; 1/500) and secondary antibody (Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120; 1/500) was used. For negative control 2 primary antibody (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291; 1/500) and secondary antibody (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077; 1/400) was used.

• 

  Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)

  All lanes: Western blot - Anti-ErbB2 / HER2 antibody [SP101] (Anti-ErbB2 / HER2 antibody [SP101] ab231438)

  All lanes: SK-BR-3 (Human breast adenocarcinoma epithelial cell) 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 0.05 µg/mL

  Predicted band size: 137 kDa

  Observed band size: 180 kDa

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)

  Immunohistochemical analysis of paraffin-embedded rat spleen tissue labeling GAPDH with ab181602 at 1/2000 dilution, followed by prediluted HRP Polymer for Rabbit/Mouse IgG. Nucleus and cytoplasmic staining on lymphocyte of rat spleen is observed. Counter stained with Hematoxylin.
  

  Negative control: Using PBS instead of primary ab, secondary ab is prediluted HRP Polymer for Rabbit/Mouse IgG.

  Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• 

  Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)

  This blot was produced using a 4-12% Bis-tris gel under the MES 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 before being incubated with Anti-GAPDH antibody ab181602 overnight at 4°C. Antibody binding was detected using the Goat Anti-Rabbit IgG H&L (IRDye^® 680RD) preadsorbed (Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed ab216777) at a 1:10,000 dilution for 1hr at room temperature and then imaged using the Licor Odyssey CLx.
  

  Image shows merged signal (red and green). Green - Anti-beta Actin antibody [AC-15] (Anti-beta Actin antibody [AC-15] ab6276)Anti-beta Actin antibody [AC-15] ab6276 observed at 42 kDa. Red - GAPDH loading control, ab181602, observed at 37 kDa.

  Lanes 1 - 2: Western blot - Anti-beta Actin antibody [AC-15] (Anti-beta Actin antibody [AC-15] ab6276) at 1/5000 dilution

  Lanes 1 - 2: Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) at 1/10000 dilution

  Lane 1: Wild-type HAP1 cell lysate (20 µg)

  Lane 2: Beta actin knockout HAP1 cell lysate (20 µg)

  Secondary

  Lanes 1 - 2: Western blot - Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed ab216772) at 1/10000 dilution

  Lanes 1 - 2: Western blot - Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed ab216777) at 1/10000 dilution

  Predicted band size: 36 kDa, 41 kDa

  Lane 1: Wild-type HAP1 cell lysate (20 μg)
  Lane 2: Beta actin knockout HAP1 cell lysate (20 μg)
  

  Lanes 1 and 2: Merged signal (red and green). Green - beta actin, Anti-beta Actin antibody [AC-15] ab6276 observed at 42 kDa. Red - loading control, ab181602 observed at 37 kDa.

  Anti-beta Actin antibody [AC-15] ab6276 was shown to specifically react with beta actin in wild-type HAP1 cells. No band was observed when beta actin knockout samples were used. Wild-type and beta actin knockout samples were subjected to SDS-PAGE. Anti-beta Actin antibody [AC-15] ab6276 (beta actin) and ab181602 (loading control to GAPDH) were diluted 1/5000 and 1/10 000 and incubated overnight at 4°C. Blots were developed with Goat anti-Mouse IgG H&L (IRDye^® 800CW) preadsorbed (Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed ab216772) and Goat Anti-Rabbit IgG H&L (IRDye^® 680RD) preadsorbed (Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed ab216777) secondary antibodies at 1/10 000 dilution for 1 h at room temperature before imaging.

• 

  Flow Cytometry (Intracellular) - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)

  Intracellular flow cytometric analysis of 2% paraformaldehyde-fixed Jurkat (Human T cell leukemia cells from peripheral blood) cells labeling GAPDH with ab181602 at 1/180 dilution (red) compared with a rabbit monoclonal IgG isotype control (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (FITC) at 1/150 dilution was used as the secondary antibody.
  

• 

  Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)

  Blocking/Dilution buffer: 5% NFDM/TBST.

  All lanes: Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) at 1/50000 dilution

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

  Lane 2: Xenopus tropicalis muscle lysates at 20 µg

  Lane 3: UMNSAH/DF-1 (Transformed chicken embyronic fibroblast cells) whole cell lysates at 20 µg

  Lane 4: Jurkat (Human T cell leukemia cells from peripheral blood) whole cell lysates at 20 µg

  Secondary

  All lanes: Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

  Predicted band size: 36 kDa

  Observed band size: 36 kDa

• 

  Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)

  Different batches of ab181602 were tested on HeLa (Human cervix adenocarcinoma epithelial cell) lysate at 1.0 µg/ml. 15 µg of lysate was loaded in each lane. Bands observed at 36 kDa.

  All lanes: Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)

  Predicted band size: 36 kDa

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)

  Immunohistochemical analysis of paraffin-embedded human transitional cell carcinoma of bladder tissue labeling GAPDH with ab181602 at 1/2000 dilution, followed by prediluted HRP Polymer for Rabbit/Mouse IgG. Cytoplasmic and nucleus staining on the tumor cells of transitional cell carcinoma of human bladder is observed. Counter stained with Hematoxylin.
  

  Negative control: Using PBS instead of primary ab, secondary ab is prediluted HRP Polymer for Rabbit/Mouse IgG.

  Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• 

  Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)

  Blocking/Dilution buffer: 5% NFDM/TBST.

  All lanes: Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) at 1/10000 dilution

  Lane 1: COS-1 (African green monkey kidney fibroblast-like cell line) whole cell lysates at 20 µg

  Lane 2: Zebrafish lysates at 20 µg

  Secondary

  All lanes: Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

  Predicted band size: 36 kDa

  Observed band size: 36 kDa

• 

  Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)

  Blocking/Dilution buffer: 5% NFDM/TBST.

  All lanes: Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) at 1/10000 dilution

  Lane 1: Human fetal brain lysates at 10 µg

  Lane 2: Human fetal heart lysates at 10 µg

  Secondary

  All lanes: Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution

  Predicted band size: 36 kDa

  Observed band size: 36 kDa

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)

  Immunohistochemical analysis of paraffin-embedded mouse spleen tissue labeling GAPDH with ab181602 at 1/2000 dilution, followed by prediluted HRP Polymer for Rabbit/Mouse IgG. Nucleus and cytoplasmic staining on lymphocytes of mouse spleen is observed. Counter stained with Hematoxylin.
  

  Negative control: Using PBS instead of primary ab, secondary ab is prediluted HRP Polymer for Rabbit/Mouse IgG.

  Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• 

  Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)

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

  Negative control: spleen, liver, testis, lung(PMID:15194045)

  The expression profile observed is consistent with differential glycosylation of this protein (PMID: 1717152).

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

  Exposure time: Lane1:15 seconds, lanes 2-6:180 seconds

  All lanes: Western blot - Anti-CEACAM5 antibody [EPR28723-86] (Anti-CEACAM5 antibody [EPR28723-86] ab317618) at 1/1000 dilution

  Lane 1: Mouse colon cancer tissue lysate at 20 µg

  Lane 2: Mouse colon tissue lysate at 20 µg

  Lane 3: Mouse spleen tissue lysate at 20 µg

  Lane 4: Mouse liver tissue lysate at 20 µg

  Lane 5: Mouse testis tissue lysate at 20 µg

  Lane 6: Mouse lung tissue 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: 106 kDa, 130-180 kDa, 36 kDa

• 

  Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)

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

  The expression of Tau (O-GlcNAc S400) is reduced following deglycosylation induced by PNGase F.

  The identity of the higher MW band at approximately 110 kDa is unknown.

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

  All lanes: Western blot - Anti-Tau (O-GlcNAc S400) antibody [EPR24891-48] (Anti-Tau (O-GlcNAc S400) antibody [EPR24891-48] ab320076) at 1/1000 dilution

  Lane 1: 293T cells transfected with a human wild-type Tau 2N4R expression vector containing a myc-His-tag® were treated with 10 uM Thiamet G for 48 hours whole cell lysate at 20 µg

  Lane 2: 293T cells transfected with a human wild-type Tau 2N4R expression vector containing a myc-His-tag® were treated with 10 uM Thiamet G for 48 hours whole cell lysate treated with Peptide:N-glycosidase F (PNGase F) 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: 25-75 kDa, 36 kDa

  Exposure time: 92s

• 

  Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)

  The identity of the bands between 25 kDa and 37 kDa are unknown.

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

  All lanes: Western blot - Anti-Squalene Epoxidase antibody [EPR29195-89] (Anti-Squalene Epoxidase antibody [EPR29195-89] ab317628) at 1/1000 dilution

  Lane 1: F9 (mouse embryonal carcinoma epithelial cell) transfected with scrambled siRNA control whole cell lysate at 20 µg

  Lane 2: F9 transfected with siRNA specifically targeting Squalene Epoxidase 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: 64 kDa, 36 kDa

  Exposure time: 62s

• 

  Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)

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

  Low expression: J774A.1(PMID: 8969198).

  The expression molecular weight observed is consistent with what has been described in the literature (PMID: 37170687).

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

  All lanes: Western blot - Anti-Semaphorin 4D/CD100 antibody [EPR26247-28] (Anti-Semaphorin 4D/CD100 antibody [EPR26247-28] ab320634) at 1/1000 dilution

  Lane 1: EL4 (mouse lymphoma T lymphocyte) whole cell lysate at 20 µg

  Lane 2: J774A.1 (mouse reticum cell sarcoma monocyte/macrophage ) 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: 120 kDa, 150 kDa, 36 kDa

  Exposure time: 180s

• 

  Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)

  Low expression: HEL, HepG2.

  The identity of the lower MW band at approximately 26 kDa (in lanes 1-3) is unknown.

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

  All lanes: Western blot - Anti-PKC epsilon antibody [EPR28194-50] (Anti-PKC epsilon antibody [EPR28194-50] ab317498) at 1/1000 dilution

  Lane 1: U-2 OS (human bone osteosarcoma epithelial cell) whole cell lysate at 20 µg

  Lane 2: HEL (human erythroleukemia erythroblast) whole cell lysate at 20 µg

  Lane 3: HepG2 (human hepatocellar carcinoma epithelial cell) 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: 85 kDa, 36 kDa

  Exposure time: 15s

• 

  Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)

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

  Low expression: colon (PMID: 8876214)

  The identity of the lower MW band at approximately 40 kDa is unknown.

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

  All lanes: Western blot - Anti-Semaphorin 4D/CD100 antibody [EPR26247-28] (Anti-Semaphorin 4D/CD100 antibody [EPR26247-28] ab320634) at 1/1000 dilution

  Lane 1: Mouse lymph node tissue lysate at 20 µg

  Lane 2: Mouse kidney tissue lysate at 20 µg

  Lane 3: Mouse hippocampus tissue lysate at 20 µg

  Lane 4: Mouse colon tissue 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: 120-150 kDa, 36 kDa

  Exposure time: 37s

• 

  Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)

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

  Negative control: placenta, spleen (PMID: 1321343).

  The molecular weight observed is consistent with what has been described in the literature (PMID: 29615706). This antibody can recognize CaMKII beta, CaMKII alpha, CaMKII delta and CaMKII gamma. They have many isofroms and isoforms have different molecular weight.

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

  Exposure time: Lanes 1, 5: 26 seconds; Lanes 2-4, 6-7: 125 seconds

  All lanes: Western blot - Anti-CAMK2B/2D/2G (phospho T287) + CAMK2A (phospho T286) antibody [EPR28075-37] (Anti-CAMK2B/2D/2G (phospho T287) + CAMK2A (phospho T286) antibody [EPR28075-37] ab320638) at 1/1000 dilution

  Lane 1: Mouse brain tissue lysate at 20 µg

  Lane 2: Mouse cerebellum tissue lysate at 20 µg

  Lane 3: Mouse placenta tissue lysate at 20 µg

  Lane 4: Mouse spleen tissue lysate at 20 µg

  Lane 5: Rat brain tissue lysate at 20 µg

  Lane 6: Rat cerebellum tissue lysate at 20 µg

  Lane 7: Rat placenta tissue 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: 50-60 kDa, 36 kDa

• 

  Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)

  The identity of the lower MW band at approximately 26 kDa (in lanes 1-7) are unknown.

  The identity of the higher MW band at approximately 240 kDa (only detected in fresh cells) is unknown.

  In lanes 1-3, the lysate was stored at -80? prior to Western Blotting. The bands between 37 kDa and 84 kDa are likely to be degradation products. In lanes 4-7, the lysates were freshly made and used for Western Blotting immediately to minimize protein degradation.

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

  All lanes: Western blot - Anti-PKC epsilon antibody [EPR28194-50] (Anti-PKC epsilon antibody [EPR28194-50] ab317498) at 1/1000 dilution

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

  Lanes 2 and 6: NIH/3T3 (mouse embryonic fibroblast) whole cell lysate at 20 µg

  Lanes 3 and 7: PC-12 (rat adrenal gland pheochromocytoma cell) whole cell lysate at 20 µg

  Lane 4: HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 20 µg

  Lane 5: 293T (human embryonic kidney epithelial cell) 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: 85 kDa, 36 kDa

  Exposure time: 180s

• 

  Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)

  Low expression: NIH:OVCAR-3 (PMID: 24386507), U-937.

  The identity of the lower MW band at approximately 40 kDa is unknown.

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

  All lanes: Western blot - Anti-ECT2 antibody [EPR29014-24] (Anti-ECT2 antibody [EPR29014-24] ab317825) at 1/1000 dilution

  Lane 1: LoVo (human colorectal adenocarcinoma epithelial cell) whole cell lysate at 20 µg

  Lane 2: HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 20 µg

  Lane 3: NIH:OVCAR-3 (human ovary adenocarcinoma epithelial cell) whole cell lysate at 20 µg

  Lane 4: U-937 (human histiocytic lymphoma monocyte) 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: 103 kDa, 36 kDa

  Exposure time: 125s

• 

  Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)

  The expression of Bid is downregulated in response to TNF alpha and BV-6 treatment (PMID: 11462023).

  The identity of the higher MW bands at approximately 250 kDa are unknown.

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

  All lanes: Western blot - Anti-Bid antibody [EPR27444-102] (Anti-Bid antibody [EPR27444-102] ab317809) at 1/1000 dilution

  Lane 1: Untreated L-929 (mouse connective tissue fibroblast) whole cell lysate at 50 µg

  Lane 2: L-929 treated with 20ng/ml TNF alpha and 10µM BV-6 for 10h whole cell lysate at 50 µg

  Lane 3: C2C12 (mouse myoblast) whole cell lysate at 50 µg

  Secondary

  All lanes: Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

  Observed band size: 22 kDa, 36 kDa

  Exposure time: 6s

• 

  Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)

  Low expression: mouse lung (PMID: 22446738).

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

  All lanes: Western blot - Anti-Bid antibody [EPR27444-102] (Anti-Bid antibody [EPR27444-102] ab317809) at 1/1000 dilution

  Lane 1: Mouse spleen tissue lysate at 50 µg

  Lane 2: Mouse thymus tissue lysate at 50 µg

  Lane 3: Mouse lung tissue lysate at 50 µg

  Secondary

  All lanes: Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

  Observed band size: 22 kDa, 36 kDa

  Exposure time: 6s

• 

  Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)

  Negative control: HeLa, JEG-3 (PMID: 2739723).

  The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID: 2739723).

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

  All lanes: Western blot - Anti-Brn-2 antibody [EPR29141-41] (Anti-Brn-2 antibody [EPR29141-41] ab317750) at 1/1000 dilution

  Lane 1: A375 (human malignant melanoma epithelial cell) whole cell lysate at 20 µg

  Lane 2: HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 20 µg

  Lane 3: JEG-3 (human placenta epithelial cell) 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: 54 kDa, 36 kDa

  Exposure time: 114s

• 

  Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)

  Negative control: heart, liver (PMID: 2739723).

  The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID: 2739723).

  The identity of the higher MW band at approximately 75 kDa (in lane 2) and the lower MW band at approximately 40 kDa (in lane 3) are unknown.

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

  All lanes: Western blot - Anti-Brn-2 antibody [EPR29141-41] (Anti-Brn-2 antibody [EPR29141-41] ab317750) at 1/1000 dilution

  Lane 1: Mouse E17 brain tissue lysate at 50 µg

  Lane 2: Mouse heart tissue lysate at 50 µg

  Lane 3: Mouse liver brain tissue lysate at 50 µg

  Lane 4: Rat P5 brain tissue lysate at 50 µg

  Lane 5: Rat liver tissue lysate at 50 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

  Developed using the ECL technique.

  Observed band size: 54 kDa, 36 kDa

  Exposure time: 8s

• 

  Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)

  Lane1 and 2 were freshly made and used for Western Blotting immediately to minimize protein degradation.

  
  
  Lanes 1 and 2 were developed using a higher sensitivity ECL substrate. The high-sensitivity ECL substrate allows for the detection of proteins in the mid-femtogram range.
  

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

  Exposure time: Lane1-2: 136 seconds, lane3: 180 seconds

  All lanes: Western blot - Anti-PNAD antibody [EPR28215-189] (Anti-PNAD antibody [EPR28215-189] ab318270) at 1/1000 dilution

  Lane 1: 293T (human embryonic kidney epithelial cell) transfected with scrambled siRNA control whole cell lysate at 20 µg

  Lane 2: 293T transfected with siRNA specifically targeting PNAD whole cell lysate at 20 µg

  Lane 3: HepG2 (human hepatocellular carcinoma epithelial cell) 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: 34 kDa, 36 kDa

• 

  Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)

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

  Low expression: skeletal muscle.

  To minimize protein degradation, tissues were lysed immediately after harvest and then applied to a gel and transfer membrane for Western blotting as soon as possible.

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

  All lanes: Western blot - Anti-TOX antibody [EPR28108-10] (Anti-TOX antibody [EPR28108-10] ab322259) at 1/1000 dilution

  Lane 1: Mouse thymus tissue lysate at 20 µg

  Lane 2: Mouse skeletal muscle tissue 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: 57 kDa, 63 kDa, 36 kDa

  Exposure time: 6s

• 

  Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)

  Low expression: heart (PMID:1556149), spleen (PMID:1556149), skeletal muscle.

  The identity of the bands between 25 kDa and 50 kDa are unknown.

  Lanes 6-11 are applied with Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 and lanes 1-5 are applied with Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000.

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

  All lanes: Western blot - Anti-PKC epsilon antibody [EPR28194-50] (Anti-PKC epsilon antibody [EPR28194-50] ab317498) at 1/1000 dilution

  Lane 1: Human hypothalamus tissue lysate at 20 µg

  Lane 2: Human cerebellum tissue lysate at 20 µg

  Lane 3: Human hippocampus tissue lysate at 20 µg

  Lane 4: Human skeletal muscle tissue lysate at 20 µg

  Lane 5: Human spleen tissue lysate at 20 µg

  Lane 6: Mouse cerebral cortex tissue lysate at 20 µg

  Lane 7: Mouse skeletal muscle tissue lysate at 20 µg

  Lane 8: Mouse heart tissue lysate at 20 µg

  Lane 9: Rat cerebral cortex tissue lysate at 20 µg

  Lane 10: Rat skeletal muscle tissue lysate at 20 µg

  Lane 11: Rat heart tissue lysate at 20 µg

  Secondary

  Lanes 1 - 5: Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

  Lanes 10, 11, 6, 7, 8 and 9: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution

  Observed band size: 85 kDa, 36 kDa

  Exposure time: 6s

• 

  Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)

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

  All lanes: Western blot - Anti-ACSL6/ACS2 antibody [EPR29174-76] (Anti-ACSL6/ACS2 antibody [EPR29174-76] ab317703) at 1/1000 dilution

  All lanes: Jurkat (human T cell leukemia T lymphocyte from peripheral blood) 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: 74 kDa, 36 kDa

  Exposure time: 180s

• 

  Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)

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

  Low expression: A-204, HT-1080, RD (PMID: 12447693).

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

  All lanes: Western blot - Anti-ID2 antibody [EPR29025-9] (Anti-ID2 antibody [EPR29025-9] ab320646) at 1/1000 dilution

  Lane 1: Untreated A-673 (human muscle ewings sarcoma cell line) whole cell lysate at 30 µg

  Lane 2: A-673 treated with 20μM MG-132 for 4 hours whole cell lysate at 30 µg

  Lane 3: Untreated HCT-15 (human colon epithelial cell) whole cell lysate at 30 µg

  Lane 4: HCT-15 treated with 20μM MG-132 for 4 hours whole cell lysate at 30 µg

  Lane 5: A-204 (human muscle rhabdomyosarcoma cell) whole cell lysate at 30 µg

  Lane 6: HT-1080 (human fibrosarcoma epithelial cell) whole cell lysate at 30 µg

  Lane 7: RD (human muscle mtinucleated spindle-shaped cell) whole cell lysate at 30 µg

  Lane 8: Untreated HCT 116 (human colorectal carcinoma epithelial cell) whole cell lysate at 30 µg

  Lane 9: HCT 116 treated with 10μM MG-132 for 4 hours whole cell lysate at 30 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: 15 kDa, 36 kDa

  Exposure time: 180s

• 

  Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)

  Low expression: liver, kidney (PMID: 12767919, PMID: 10502316).

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

  All lanes: Western blot - Anti-ACSL6/ACS2 antibody [EPR29174-76] (Anti-ACSL6/ACS2 antibody [EPR29174-76] ab317703) at 1/1000 dilution

  Lane 1: Rat brain tissue lysate at 20 µg

  Lane 2: Rat liver tissue lysate at 20 µg

  Lane 3: Mouse brain tissue lysate at 20 µg

  Lane 4: Mouse hypothalamus tissue lysate at 20 µg

  Lane 5: Mouse kidney tissue 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: 74 kDa, 36 kDa

  Exposure time: 180s

• 

  Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)

  Negative control: HepG2 (PMID: 20093481).

  Low expression: LoVo (PMID: 22701813).

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

  All lanes: Western blot - Anti-FGFR2IIIb/FGFR2b antibody [EPR28355-27] (Anti-FGFR2IIIb/FGFR2b antibody [EPR28355-27] ab318274) at 1/1000 dilution

  Lane 1: KATO III (human stomach spherical) whole cell lysate at 20 µg

  Lane 2: HepG2 (human hepatocellular carcinoma epithelial cell) whole cell lysate at 20 µg

  Lane 3: LoVo (human colorectal adenocarcinoma epithelial cell) 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: 85-135 kDa, 36 kDa

  Exposure time: 48s

• 

  Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)

  Low expression: liver, lung (PMID: 12767919, PMID: 10502316).

  The identity of the band at approximately 37 kDa (in lane 4) is unknown.

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

  All lanes: Western blot - Anti-ACSL6/ACS2 antibody [EPR29174-76] (Anti-ACSL6/ACS2 antibody [EPR29174-76] ab317703) at 1/1000 dilution

  Lane 1: Human hypothalamus tissue lysate at 50 µg

  Lane 2: Human cerebellum tissue lysate at 50 µg

  Lane 3: Human lung tissue lysate at 50 µg

  Lane 4: Human liver tissue lysate at 50 µg

  Secondary

  All lanes: Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

  Observed band size: 74 kDa, 36 kDa

  Exposure time: 180s

• 

  Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)

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

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

  All lanes: Western blot - Anti-EXOSC10/RRP6 antibody [EPR28902-62] (Anti-EXOSC10/RRP6 antibody [EPR28902-62] ab319992) at 1/1000 dilution

  Lane 1: HeLa (human cervical adenocarcinoma epithelial cell) transfected with scrambled siRNA control whole cell lysate at 20 µg

  Lane 2: Hela transfected with siRNA specifically targeting EXOSC10/RRP6 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: 100 kDa, 36 kDa

  Exposure time: 48s

• 

  Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)

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

  Low expression: lung, skeletal muscle, spleen (PMID: 12963731)

  Negative control: liver (PMID: 12963731)

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

  All lanes: Western blot - Anti-ATP6V0D2 antibody [EPR29173-521] (Anti-ATP6V0D2 antibody [EPR29173-521] ab321809) at 1/1000 dilution

  Lane 1: Human kidney tissue lysate at 40 µg

  Lane 2: Human liver tissue lysate at 40 µg

  Lane 3: Human lung tissue lysate at 40 µg

  Lane 4: Mouse kidney tissue lysate at 40 µg

  Lane 5: Mouse skeletal muscle tissue lysate at 40 µg

  Lane 6: Mouse spleen tissue lysate at 40 µg

  Lane 7: Mouse lung tissue lysate at 40 µg

  Secondary

  All lanes: Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

  Observed band size: 40 kDa, 36 kDa

  Exposure time: 37s

• 

  Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602)

  Low expression: skeletal muscle (PMID: 16571602).

  Anti-MCRS1/MSP58 antibody [EPR27229-81] ab316109 detects isoform of MCRS1, around 40 kDa in Western blot (PMID: 25816988).

  Lanes 1-3 are applied with Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000; Lanes 4-9 are applied with Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000.

  The identity of the band at around 20 kDa (in lane 1 and 2) is unknown.

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

  All lanes: Western blot - Anti-MCRS1/MSP58 antibody [EPR27229-81] (Anti-MCRS1/MSP58 antibody [EPR27229-81] ab316109) at 1/1000 dilution

  Lane 1: Human testis tissue lysate at 20 µg

  Lane 2: Human spleen tissue lysate at 20 µg

  Lane 3: Human skeletal muscle tissue lysate at 20 µg

  Lane 4: Mouse testis tissue lysate at 20 µg

  Lane 5: Mouse spleen tissue lysate at 20 µg

  Lane 6: Mouse skeletal muscle tissue lysate at 20 µg

  Lane 7: Rat testis tissue lysate at 20 µg

  Lane 8: Rat spleen tissue lysate at 20 µg

  Lane 9: Rat skeletal muscle tissue lysate at 20 µg

  Secondary

  Lanes 1 - 3: Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG (Merck DC03L) at 1/2000 dilution

  Lanes 4 - 9: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution

  Observed band size: 52 kDa, 36 kDa

  Exposure time: 180s