Anti-beta Actin 抗体 [mAbcam 8226] - Loading Control

• WB

Unknown

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 5% Milk before being incubated with ab8226 overnight at 4°C. Antibody binding was detected using Goat Anti-Mouse IgG H&L (Alexa Fluor® 790) (ab175783) at a 1 : 10,000 dilution for 1hr at room temperature and then imaged using the Licor Odyssey CLx.

All lanes:

Western blot - Anti-beta Actin antibody [mAbcam 8226] - Loading Control (ab8226) at 1/1000 dilution

Lane 1:

A431 (Human epidermoid carcinoma cell line) Whole Cell Lysate at 20 µg

Lane 2:

HEK293 (Human embryonic kidney cell line) Whole Cell Lysate at 20 µg

Lane 3:

NIH 3T3 (Mouse embryo fibroblast cell line) Whole Cell Lysate at 20 µg

Lane 4:

PC12 (Rat adrenal gland pheochromocytoma cell line) Whole Cell Lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Mouse IgG H&L (Alexa Fluor® 790) (<a href='/products/secondary-antibodies/goat-mouse-igg-h-l-alexa-fluor-790-ab175783'>ab175783</a>) at 1/10000 dilution

Predicted band size: 36 kDa,41 kDa

Observed band size: 42 kDa

false

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-beta Actin antibody [mAbcam 8226] - Loading Control (AB8226)

Immunohistochemical analysis of formalin fixed paraffin embedded human colon labelling beta actin with ab8226 at a concentration of 0.06 µg/ml. The immunostaining was performed on a Ventana DISCOVERY ULTRA (Roche Tissue Diagnostics) instrument with an OptiView DAB IHC Detection Kit. Heat mediated antigen retrieval was conducted for 32 mins at 100°C with ULTRA cell conditioning solution (CC1, pH 8.5). ab8226 anti-beta actin antibody [mAbcam 8226] was incubated at 37°C for 16 mins. Sections were counterstained with Hematoxylin II. Image inset shows absence of staining in secondary antibody only control.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-beta Actin antibody [mAbcam 8226] - Loading Control (AB8226)

IHC image of ab8226 staining beta Actin in human colon formalin fixed paraffin embedded tissue sections*, performed on a Leica Bond. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab8226, 0.1μg/ml working concentration, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. No primary antibody was used in the secondary only control (shown on the inset).
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-beta Actin antibody [mAbcam 8226] - Loading Control (AB8226)

This data was developed using the same antibody clone in a different buffer formulation containing only PBS (ab264083). ab264083 staining beta Actin in HeLa cells. The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% PBS-Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab264083 at 5µg/ml and ab6046, Rabbit polyclonal to beta Tubulin - Loading Control. Cells were then incubated with ab150117, Goat polyclonal Secondary Antibody to Mouse IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (shown in green) and ab150080, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 594) at 1/1000 dilution (shown in pseudocolour magenta). Nuclear DNA was labelled with DAPI (shown in blue). Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-beta Actin antibody [mAbcam 8226] - Loading Control (AB8226)

IHC image of ab8226 staining beta Actin in rat colon formalin fixed paraffin embedded tissue sections, performed on a Leica Bond. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab8226, 0.5μg/ml working concentration, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. No primary antibody was used in the secondary only control (shown on the inset).
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

• WB

Lab

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 ab205719, 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 - Goat Anti-Mouse IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-mouse-igg-h-l-hrp-ab205719'>ab205719</a>) at 1/5000 dilution

Predicted band size: 36 kDa

Observed band size: 42 kDa

true

Exposure time: 10s

• WB

Unknown

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

Predicted band size: 41 kDa

Observed band size: 42 kDa

true

Exposure time: 10s

• WB

Lab

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) (<a href='/products/secondary-antibodies/donkey-mouse-igg-h-l-hrp-ab205724'>ab205724</a>) at 1/2000 dilution

Predicted band size: 36 kDa

Observed band size: 42 kDa

true

Exposure time: 15s

• WB

Project

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

Predicted band size: 41 kDa

Observed band size: 42 kDa

false

Exposure time: 8min

• WB

Lab

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:

<a href='/products/secondary-antibodies/donkey-mouse-igg-h-l-hrp-ab205724'>ab205724</a> (Left Image) at 1/2000 and a competitor secondary (Right Image) at 1/2000. Notice the increased background of the competitor product.

Predicted band size: 36 kDa

Observed band size: 42 kDa

false

Exposure time: 15s

• WB

Unknown

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) (<a href='/products/secondary-antibodies/rabbit-mouse-igg-h-l-alkaline-phosphatase-ab97043'>ab97043</a>) at 1/5000 dilution

Lanes 3 - 4:

Western blot - Rabbit Anti-Mouse IgG H&L (Alkaline Phosphatase) (<a href='/products/secondary-antibodies/rabbit-mouse-igg-h-l-alkaline-phosphatase-ab97043'>ab97043</a>) at 1/20000 dilution

Lanes 5 - 6:

Western blot - Rabbit Anti-Mouse IgG H&L (Alkaline Phosphatase) (<a href='/products/secondary-antibodies/rabbit-mouse-igg-h-l-alkaline-phosphatase-ab97043'>ab97043</a>) at 1/50000 dilution

Predicted band size: 36 kDa

true

Exposure time: 20min

• WB

Unknown

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) (<a href='/products/secondary-antibodies/rabbit-mouse-igg-h-l-hrp-ab6728'>ab6728</a>) at 1/5000 dilution

Predicted band size: 41 kDa

false

Exposure time: 10s

• WB

Unknown

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 - Goat Anti-Mouse IgG H&L (Alkaline Phosphatase) (<a href='/products/secondary-antibodies/goat-mouse-igg-h-l-alkaline-phosphatase-ab97020'>ab97020</a>) at 1/5000 dilution

Lanes 3 - 4:

Western blot - Goat Anti-Mouse IgG H&L (Alkaline Phosphatase) (<a href='/products/secondary-antibodies/goat-mouse-igg-h-l-alkaline-phosphatase-ab97020'>ab97020</a>) at 1/20000 dilution

Lanes 5 - 6:

Western blot - Goat Anti-Mouse IgG H&L (Alkaline Phosphatase) (<a href='/products/secondary-antibodies/goat-mouse-igg-h-l-alkaline-phosphatase-ab97020'>ab97020</a>) at 1/50000 dilution

Predicted band size: 36 kDa

true

Exposure time: 4min