Western blot - Anti-beta Actin antibody [mAbcam 8226] - Loading Control (ab8226)
This blot was produced using a 4-12% 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 2% Bovine Serum Albumin before being incubated with ab8226 overnight at 4°C. Antibody binding was detected using ab205724, and visualised using ECL development solution ab133406.
All lanes:
Western blot - Anti-beta Actin antibody [mAbcam 8226] - Loading Control (ab8226) at 1 µg/mL
Lane 1:
Liver (Human) Tissue Lysate at 10 µg
Lane 2:
Liver (Mouse) Tissue Lysate at 10 µg
Lane 3:
Liver (Rat) Tissue Lysate at 10 µg
Lane 4:
HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 10 µg
Lane 5:
NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate at 10 µg
Lane 6:
PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell Lysate at 10 µg
Secondary
All lanes:
Western blot - Donkey Anti-Mouse IgG H&L (HRP) (ab205724) at 1/2000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 36 kDa
Observed band size: 42 kDa
Exposure time: 15s
Western blot - Anti-beta Actin antibody [mAbcam 8226] - Loading Control (ab8226)
Lane 1:
Western blot - Anti-beta Actin antibody [mAbcam 8226] - Loading Control (ab8226) at 1/1000 dilution
Lane 2:
Western blot - Anti-beta Actin antibody [mAbcam 8226] - Loading Control (ab8226) at 1/10000 dilution
Lanes 3 - 4:
Western blot - Anti-beta Actin antibody [mAbcam 8226] - Loading Control (ab8226) at 1/500 dilution
Lanes 1 - 2:
HeLa (Human epithelial cell line from cervix adenocarcinoma) cell lysate at 20 µg
Lane 3:
HEK293 (Human epithelial cell line from embryonic kidney) cell lysate at 20 µg
Lane 4:
NIH/3T3 (Mouse embryo fibroblast cell line) cell lysate at 20 µg
Secondary
All lanes:
Western blot - Rabbit Anti-Mouse IgG H&L (HRP) (ab6728) at 1/5000 dilution
Performed under reducing conditions.
Predicted band size: 41 kDa
Exposure time: 10s
Western blot - Anti-beta Actin antibody [mAbcam 8226] - Loading Control (ab8226)
Western blot image using the Optiblot Reducing Electrophoresis Kit - 10 x 10 cm (4-20%) with the Prism Ultra Protein Ladder (ab116028) 5μl used. We recommend using our ECL substrate kit (ab65623).
All lanes:
Western blot - Anti-beta Actin antibody [mAbcam 8226] - Loading Control (ab8226) at 1 µg/mL
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
Lane 3:
A431 (Human epidermoid carcinoma cell line) whole cell lysate at 20 µg
Lane 4:
HEK293 (Human epithelial cell line from embryonic kidney) whole cell lysate at 20 µg
Lane 5:
HepG2 (Human liver hepatocellular carcinoma cell line) whole cell lysate at 20 µg
Secondary
All lanes:
HRP-conjugated goat anti-mouse IgG at 1/5000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 41 kDa
Observed band size: 42 kDa
Exposure time: 10s
Western blot - Anti-beta Actin antibody [mAbcam 8226] - Loading Control (ab8226)
All lanes:
Western blot - Anti-beta Actin antibody [mAbcam 8226] - Loading Control (ab8226) at 1 µg/mL
Lanes 1 and 4:
HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 10 µg
Lanes 2 and 5:
Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate at 10 µg
Lanes 3 and 6:
NIH/3T3 (Mouse embryo fibroblast cell line) whole cell lysate at 10 µg
Secondary
All lanes:
Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
Performed under reducing conditions.
Predicted band size: 41 kDa
Observed band size: 42 kDa
Exposure time: 8min
Western blot - Anti-beta Actin antibody [mAbcam 8226] - Loading Control (ab8226)
All lanes:
Western blot - Anti-beta Actin antibody [mAbcam 8226] - Loading Control (ab8226) at 1 µg/mL
All lanes:
HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 10 µg
Secondary
Lanes 1 - 2:
Western blot - Rabbit Anti-Mouse IgG H&L (Alkaline Phosphatase) (ab97043) at 1/5000 dilution
Lanes 3 - 4:
Western blot - Rabbit Anti-Mouse IgG H&L (Alkaline Phosphatase) (ab97043) at 1/20000 dilution
Lanes 5 - 6:
Western blot - Rabbit Anti-Mouse IgG H&L (Alkaline Phosphatase) (ab97043) at 1/50000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 36 kDa
Exposure time: 20min
Western blot - Anti-beta Actin antibody [mAbcam 8226] - Loading Control (ab8226)
This blot was produced using a 4-12% 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 2% Bovine Serum Albumin before being incubated with ab8226 overnight at 4°C. Antibody binding was detected using ab205724 (Left Image) and a competitor secondary (Right Image), and visualised using ECL development solution ab133406.
All lanes:
Western blot - Anti-beta Actin antibody [mAbcam 8226] - Loading Control (ab8226) at 1 µg/mL
Lane 1:
Liver (Human) Tissue Lysate at 10 µg
Lane 2:
Liver (Mouse) Tissue Lysate at 10 µg
Lane 3:
Liver (Rat) Tissue Lysate at 10 µg
Secondary
All lanes:
ab205724 (Left Image) at 1/2000 and a competitor secondary (Right Image) at 1/2000. Notice the increased background of the competitor product.
Performed under reducing conditions.
Predicted band size: 36 kDa
Observed band size: 42 kDa
Exposure time: 15s
