Anti-beta Actin antibody - Loading Control

19 product images

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  Western blot - Anti-beta Actin antibody - Loading Control (ab8227)

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

  Lanes 1-3: Blocked with 2% BSA
  

  Lanes 4-6: Blocked with 3% Milk

  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 or 3% Milk before being incubated with ab8227 overnight at 4°C. Antibody binding was detected using an anti rabbit HRP secondary antibody, and visualised using ECL development solution ECL Substrate Kit (High Sensitivity) ab133406

  All lanes: Western blot - Anti-beta Actin antibody - Loading Control (ab8227) at 1 µg/mL

  Lane 1: HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate (blocked with 2% BSA) at 10 µg

  Lane 2: NIH/3T3 (mouse embryo fibroblast cell line) whole cell lysate (blocked with 2% BSA) at 10 µg

  Lane 3: Rat Liver tissue lysate (blocked with 2% BSA) at 10 µg

  Lane 4: HeLa whole cell lysate (blocked with 3% Milk) at 10 µg

  Lane 5: NIH/3T3 whole cell lysate (blocked with 3% Milk) at 10 µg

  Lane 6: Rat Liver tissue lysate (blocked with 3% Milk) at 10 µg

  Secondary

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

  Developed using the ECL technique.

  Performed under reducing conditions.

  Predicted band size: 41 kDa

  Observed band size: 45 kDa

  Exposure time: 10s

• 

  Western blot - Anti-beta Actin antibody - Loading Control (ab8227)

  Beta Actin Western blot staining of HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate using rabbit Anti-beta Actin antibody

  All lanes: Western blot - Anti-beta Actin antibody - Loading Control (ab8227) at 1 µg/mL

  All lanes: HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 10 µg

  Secondary

  Lanes 1 - 2: Western blot - Donkey Anti-Rabbit IgG H&L (Alkaline Phosphatase) (Donkey Anti-Rabbit IgG H&L (Alkaline Phosphatase) ab97061) at 1/5000 dilution

  Lanes 3 - 4: Western blot - Donkey Anti-Rabbit IgG H&L (Alkaline Phosphatase) (Donkey Anti-Rabbit IgG H&L (Alkaline Phosphatase) ab97061) at 1/20000 dilution

  Lanes 5 - 6: Western blot - Donkey Anti-Rabbit IgG H&L (Alkaline Phosphatase) (Donkey Anti-Rabbit IgG H&L (Alkaline Phosphatase) ab97061) at 1/50000 dilution

  Developed using the ECL technique.

  Performed under reducing conditions.

  Predicted band size: 36 kDa

  Exposure time: 3min

• 

  Western blot - Anti-beta Actin antibody - Loading Control (ab8227)

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

  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 ab8227 overnight at 4°C. Antibody binding was detected using Donkey Anti-Rabbit IgG H&L (HRP) ab205722, and visualised using ECL development solution ECL Substrate Kit (High Sensitivity) ab133406.

  All lanes: Western blot - Anti-beta Actin antibody - Loading Control (ab8227) 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-Rabbit IgG H&L (HRP) (Donkey Anti-Rabbit IgG H&L (HRP) ab205722) at 1/10000 dilution

  Developed using the ECL technique.

  Performed under reducing conditions.

  Predicted band size: 36 kDa

  Observed band size: 42 kDa

  Exposure time: 10s

• 

  Western blot - Anti-beta Actin antibody - Loading Control (ab8227)

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

  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 ab8227 overnight at 4°C. Antibody binding was detected using Goat Anti-Rabbit IgG H&L (HRP) ab205718, and visualised using ECL development solution ECL Substrate Kit (High Sensitivity) ab133406.

  All lanes: Western blot - Anti-beta Actin antibody - Loading Control (ab8227) 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-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab205718) at 1/50000 dilution

  Developed using the ECL technique.

  Performed under reducing conditions.

  Predicted band size: 36 kDa

  Observed band size: 42 kDa

  Exposure time: 30s

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-beta Actin antibody - Loading Control (ab8227)

  Beta Actin Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) staining using rabbit Anti-beta Actin antibody

  IHC image of beta actin staining in a section of formalin-fixed paraffin-embedded normal human colon*. The section was pre-treated using pressure cooker heat mediated antigen retrieval with sodium citrate buffer (pH6) for 30mins. The section was then incubated with ab8227, 1/1000 dilution, for 15 mins at room temperature. A goat anti-rabbit biotinylated secondary antibody (Goat Anti-Rabbit IgG H&L (Biotin) ab6720, 1/1000 dilution) was used to detect the primary, and visualized using an HRP conjugated ABC system. Streptavidin HRP was used, Streptavidin (HRP) ab7403 at a 1/10000 dilution. DAB was used as the chromogen (ab103723), diluted 1/100 and incubated for 10min at room temperature. The section was then counterstained with haematoxylin and mounted with DPX.
  

  The inset negative control image is taken from an identical assay without primary antibody.

  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

• 

  Western blot - Anti-beta Actin antibody - Loading Control (ab8227)

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

  All lanes: Western blot - Anti-beta Actin antibody - Loading Control (ab8227) at 1/1000 dilution

  Lanes 1 and 6: HeLa nuclear lysate at 20 µg

  Lanes 2 and 7: HeLa whole cell lysate at 20 µg

  Lanes 3 and 8: A431 cell lysate at 20 µg

  Lane 4: Jurkat cell lysate at 20 µg

  Lanes 5 and 10: HEK 293 cell lysate at 20 µg

  Lane 9: Jurkate cell lysate at 20 µg

  Secondary

  All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab6721) at 1/5000 dilution

  Predicted band size: 41 kDa

  Observed band size: 41.7 kDa

  Exposure time: 5s

• 

  Western blot - Anti-beta Actin antibody - Loading Control (ab8227)

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

  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 ab8227 overnight at 4°C. Antibody binding was detected using Goat Anti-Rabbit IgG H&L (Alexa Fluor® 790) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 790) ab175781) secondary antibody 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 - Loading Control (ab8227) at 1/1000 dilution

  Lane 1: A431 (human epidermoid carcinoma cell line) Whole Cell Lysate at 20 µg

  Lane 2: HEK-293 (human epithelial cell line from embryonic kidney) Whole Cell Lysate at 20 µg

  Lane 3: NIH/3T3 (mouse embryo fibroblast cell line) Whole Cell Lysate at 20 µg

  Lane 4: PC-12 (rat adrenal gland pheochromocytoma cell line) Whole Cell Lysate at 20 µg

  Secondary

  All lanes: Western blot - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 790) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 790) ab175781) at 1/10000 dilution

  Predicted band size: 41 kDa

  Observed band size: 42 kDa

• 

  Immunocytochemistry/ Immunofluorescence - Anti-beta Actin antibody - Loading Control (ab8227)

  Beta Actin Immunocytochemistry/ Immunofluorescence staining using rabbit Anti-beta Actin antibody

  ab8227 staining beta Actin in SV40LT-SMC (rat aortic smooth muscle cells transfected with SV40). The cells were fixed with 4% formaldehyde (10min), permeabilized with 0.1% 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 ab8227 at 1μg/ml (detected with Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081, Alexa Fluor® 488 Goat anti-Rabbit, 1μg/ml, shown in green); and Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889, Mouse monoclonal [DM1A] to alpha Tubulin - Microtubule Marker (Alexa Fluor® 594), at 1/250 dilution (shown in red). Nuclear DNA was labelled with DAPI (shown in blue).
  Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-beta Actin antibody - Loading Control (ab8227)

  Beta Actin Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) staining using rabbit Anti-beta Actin antibody

  IHC image of ab8227 staining beta Actin in rat small intestine formalin fixed paraffin embedded tissue sections, performed on a Leica Bond. The section was pre-treated using heat mediated antigen retrieval with EDTA (epitope retrieval solution 2) for 20 mins. The section was then incubated with ab8227, 0.2μg/ml, 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.
  
  

• 

  Immunocytochemistry/ Immunofluorescence - Anti-beta Actin antibody - Loading Control (ab8227)

  Beta Actin Immunocytochemistry/ Immunofluorescence staining using rabbit Anti-beta Actin antibody

  ab8227 staining beta Actin in NIH/3T3 (mouse embryo fibroblast cell line) cells. The cells were fixed with 100% methanol (5min), permeabilized with 0.1% 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 ab8227 at 1μg/ml (shown in green) and Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889, Mouse monoclonal [DM1A] to alpha Tubulin - Microtubule Marker (Alexa Fluor® 594), at 1/250 dilution (shown in red). Nuclear DNA was labelled with DAPI (shown in blue).
  Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

• 

  Western blot - Anti-beta Actin antibody - Loading Control (ab8227)

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

  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 ab8227 overnight at 4°C. Antibody binding was detected using an anti-rabbit HRP secondary antibody (Goat Anti-Rabbit IgG H&L (HRP) ab6721), and visualised using ECL development solution ECL Substrate Kit (High Sensitivity) ab133406.

  All lanes: Western blot - Anti-beta Actin antibody - Loading Control (ab8227) at 0.1 µg/mL

  Lane 1: HeLa (human epithelial cell line from cervix adenocarcinoma) Whole Cell Lysate at 10 µg

  Lane 2: NIH/3T3 (mouse embryonic fibroblast cell line) Whole Cell Lysate at 10 µg

  Lane 3: Liver (Rat) Tissue Lysate at 10 µg

  Secondary

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

  Performed under reducing conditions.

  Predicted band size: 41 kDa

  Observed band size: 42 kDa

  Exposure time: 1min

• 

  Western blot - Anti-beta Actin antibody - Loading Control (ab8227)

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

  All lanes: Western blot - Anti-beta Actin antibody - Loading Control (ab8227) at 1/1000 dilution

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

  Lanes 2 and 7: HeLa whole cell lysate at 20 µg

  Lane 3: A431 (human epidermoid carcinoma cell line) cell lysate at 20 µg

  Lane 4: Jurkat (human T cell leukemia cell line from peripheral blood) cell lysate at 20 µg

  Lane 5: HEK-293 (human epithelial cell line from embryonic kidney) cell lysate at 20 µg

  Lane 6: HeLa nuclear lysate at 20 µg

  Lane 8: A431 cell lysate at 20 µg

  Lane 9: Jurkat cell lysate at 20 µg

  Lane 10: HEK-293 cell lysate at 20 µg

  Secondary

  All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab6721) at 1/5000 dilution

  Predicted band size: 41 kDa

  Observed band size: 41.7 kDa

  Exposure time: 5s

• 

  Western blot - Anti-beta Actin antibody - Loading Control (ab8227)

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

  All lanes: Western blot - Anti-beta Actin antibody - Loading Control (ab8227) at 1/1000 dilution

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

  Lane 2: NIH/3T3 (Mouse embryonic fibroblast cell line) cells at 20 µg

  Lane 3: Fish Liver at 20 µg

  Lane 4: Rabbit Liver at 20 µg

  Lane 5: MDCK (Canine kidney cell line) cells at 20 µg

  Lane 6: EBTr (cow trachea) cells at 20 µg

  Lane 7: SL-29 (chicken day 11 embryo) cells at 20 µg

  Lane 8: CHO (Chinese hamster ovary cell line) cells at 20 µg

  Lane 9: Xenopus laevis embryo at 20 µg

  Secondary

  All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab6721) at 1/5000 dilution

  Predicted band size: 41 kDa

  Observed band size: 41.7 kDa

  Exposure time: 30s

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-beta Actin antibody - Loading Control (ab8227)

  Beta Actin Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) staining using rabbit Anti-beta Actin antibody

  IHC image of beta Actin staining in normal human colon, formalin-fixed and paraffin-embedded tissue*. The section was pre-treated using pressure cooker heat mediated antigen retrieval with sodium citrate buffer (pH6) for 30mins. The section was incubated with ab8227, 3μg/ml overnight at +4°C. A anti-rabbit HRP secondary antibody (Goat Anti-Rabbit IgG Fc (HRP) ab97200, 1/200 dilution) was used for 1hr at room temperature. The section was counterstained with haematoxylin and mounted with DPX.
  

  The inset negative control image is taken from an identical assay without primary antibody.

  *Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre

• 

  Western blot - Anti-beta Actin antibody - Loading Control (ab8227)

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

  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 ab8227 overnight at 4°C. Antibody binding was detected using Goat Anti-Rabbit IgG H&L (HRP) ab205718 (Left Image) and a competitor secondary (Right Image), and visualised using ECL development solution ECL Substrate Kit (High Sensitivity) ab133406.

  All lanes: Western blot - Anti-beta Actin antibody - Loading Control (ab8227) at 1 µg/mL

  Lane 1: Liver (Mouse) Tissue Lysate at 10 µg

  Lane 2: Liver (Rat) Tissue Lysate at 10 µg

  Secondary

  All lanes: Goat Anti-Rabbit IgG H&L (HRP) ab205718 (Left Image) at 1/20,000 and a competitor secondary (Right Image) at 1/50,000. Notice the increased background of the competitor product.

  Performed under reducing conditions.

  Predicted band size: 36 kDa

  Observed band size: 42 kDa

  Exposure time: 5s

• 

  Western blot - Anti-beta Actin antibody - Loading Control (ab8227)

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

  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 ab8227 overnight at 4°C. Antibody binding was detected using Donkey Anti-Rabbit IgG H&L (HRP) ab205722 (Left Image) and a competitor secondary (Right Image), and visualised using ECL development solution ECL Substrate Kit (High Sensitivity) ab133406.

  All lanes: Western blot - Anti-beta Actin antibody - Loading Control (ab8227) at 1 µg/mL

  Lane 1: Liver (Mouse) Tissue Lysate at 10 µg

  Lane 2: Liver (Rat) Tissue Lysate at 10 µg

  Secondary

  All lanes: Donkey Anti-Rabbit IgG H&L (HRP) ab205722 (Left Image) at 1/10,000 and a competitor secondary (Right Image) at 1/10,000. Notice the decreased signal of the competitor product.

  Performed under reducing conditions.

  Predicted band size: 36 kDa

  Observed band size: 42 kDa

  Exposure time: 10s

• 

  Western blot - Anti-beta Actin antibody - Loading Control (ab8227)

  Beta Actin Western blot staining of HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate using rabbit Anti-beta Actin antibody

  Lanes 1 - 6: Western blot - Anti-beta Actin antibody - Loading Control (ab8227) at 1 µg/mL

  Lanes 1 - 2: Western blot - Anti-GNAT2 antibody (Anti-GNAT2 antibody ab97501) at 1/2000 dilution

  All lanes: HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 10 µg

  Secondary

  Lanes 3 - 4: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/10000 dilution

  Lanes 5 - 6: 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.

  Performed under reducing conditions.

  Predicted band size: 36 kDa

  Exposure time: 10s

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-beta Actin antibody - Loading Control (ab8227)

  Beta Actin Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) staining of human colon using rabbit Anti-beta Actin antibody

  Immunohistochemical analysis of formalin fixed paraffin embedded human colon labelling beta actin with ab8227 at a concentration of 2 µg/ml. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument with a Bond™ Polymer Refine Detection kit. Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution 2) for 20 mins. ab8227 anti-beta actin antibody was incubated for 30 mins at room temperature. Sections were counterstained with Hematoxylin. Image inset shows absence of staining in secondary antibody only control.

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-beta Actin antibody - Loading Control (ab8227)

  Beta Actin Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) staining of human colon using rabbit Anti-beta Actin antibody

  Immunohistochemical analysis of formalin fixed paraffin embedded human colon labelling beta actin with ab8227 at a concentration of 1 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). ab8227 anti-beta actin antibody 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.