Anti-beta Actin antibody [AC-15] - Loading Control

6 product images

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  Western blot - Anti-beta Actin antibody [AC-15] - Loading Control (ab6276)

  Beta Actin Western blot staining using mouse Anti-beta Actin antibody

  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, ab6276 observed at 42 kDa. Red - loading control, Anti-GAPDH antibody [EPR16891] - Loading Control ab181602 observed at 37 kDa.

  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. ab6276 (beta actin) and Anti-GAPDH antibody [EPR16891] - Loading Control 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.

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

  Lanes 1 - 2: Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (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

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  This image is courtesy of a customer review submitted by Dr Mark Elliott

  Western blot - Anti-beta Actin antibody [AC-15] - Loading Control (ab6276)

  Beta Actin Western blot staining using mouse Anti-beta Actin antibody

  Western Blot of ab6276 (used as loading control) with whole tissue lysate of human grey matter from BA20 (temporal cortex). ab6276 was diluted 1/50000 and incubated with the sample for 16 hours at 4°C. 5 μg of lysate was loaded onto the gel, which was blocked with 5% milk for 1 hour at 20°C. An Alexa Fluor® 680 conjugated goat anti-mouse antibody, diluted 1/5000, was used as the secondary.
  

  Bands below actin in image are synaptophysin (SYN).

  All lanes: Western blot - Anti-beta Actin antibody [AC-15] - Loading Control (ab6276)

  Predicted band size: 41 kDa

  Observed band size: 42 kDa

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  Immunocytochemistry/ Immunofluorescence - Anti-beta Actin antibody [AC-15] (ab6276)

  ab6276 staining beta Actin in SV40LT-SMC cells. The cells were fixed with 4% formaldehyde (10 min), permeabilized in 0.1% Triton X-100 for 5 minutes and then blocked in 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab6276 at a working concentration of 5μg/ml and Alexa Fluor® 647 Anti-alpha Tubulin antibody [EP1332Y] - Microtubule Marker ab190573, Rabbit monoclonal [EP1332Y] to alpha Tubulin (Alexa Fluor® 647, shown in red) at 1/250 overnight at +4°C, followed by a further incubation at room temperature for 1h with an anti-mouse AlexaFluor® 488 (Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed ab150117) at 2 μg/ml (shown in green). Nuclear DNA was labelled in blue with DAPI.
  Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

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  Western blot - Anti-beta Actin antibody [AC-15] - Loading Control (ab6276)

  Beta Actin Western blot staining using mouse Anti-beta Actin antibody

  All lanes: Western blot - Anti-beta Actin antibody [AC-15] - Loading Control (ab6276) at 1 µg/mL

  Lane 1: HeLa (human epithelial cell line from cervix adenocarcinoma) cell lysate

  Lane 2: Jurkat (human T cell leukemia cell line from peripheral blood) cell lysate

  Lane 3: COS-7 (african green monkey kidney fibroblast-like cell line) cell lysate

  Lane 4: NIH/3T3 (mouse embryonic fibroblast cell line) cell lysate

  Lane 5: PC-12 (rat adrenal gland pheochromocytoma cell line) cell lysate

  Lane 6: Rat2 (rat fibroblast cell line) cell lysate

  Lane 7: CHO (chinese hamster ovary cell line) cell lysate

  Lane 8: MDBK (bovine kidney cell line) cell lysate

  Lane 9: MDCK (canine kidney cell line) cell lysate

  Secondary

  All lanes: Goat Anti-Mouse IgG-Peroxidase

  Developed using the ECL technique.

  Predicted band size: 41 kDa

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  Western blot - Anti-beta Actin antibody [AC-15] - Loading Control (ab6276)

  Beta Actin Western blot staining using mouse Anti-beta Actin antibody

  All lanes: Western blot - Anti-beta Actin antibody [AC-15] - Loading Control (ab6276) at 1/5000 dilution

  Lane 1: HeLa nuclear at 20 µg

  Lane 2: HeLa whole cell lysate at 20 µg

  Lane 3: A431 cell lysate at 20 µg

  Lane 4: Jurkat cell lysate at 20 µg

  Lane 5: HEK293 cell lysate at 20 µg

  Secondary

  All lanes: Alexa Fluor anti mouse at 1/5000 dilution

  Performed under reducing conditions.

  Predicted band size: 41 kDa

  Observed band size: 42 kDa

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  This image is courtesy of David Grimm, Yale University, USA.

  Western blot - Anti-beta Actin antibody [AC-15] (ab6276)

  MDCK cells induced with increasing amounts of doxycycline to control expression of the gene of interest. All cells were normalized for loading with an albumin protein standard assay. Anti-beta actin (ab6276) was used at a concentration of 1:5000 in a milk blocking solution. B-actin blotting confirms the albumin assay in showing that an equal amount of lysate was loaded in each lane.

  All lanes: Western blot - Anti-beta Actin antibody [AC-15] (ab6276)

  Predicted band size: 41 kDa