Anti-beta Actin antibody is a rabbit polyclonal antibody that is used to detect beta Actin in ICC/IF, IHC-P, Western blot. Suitable for Chicken, Chinese hamster, Cow, Dog, Fish, Human, Mouse, Rabbit, Rat, Xenopus laevis samples.
- Cited in over 4,000 publications
- Trusted since 2003
pH: 7.4
Preservative: 0.02% Sodium azide
Constituents: 98.98% PBS, 1% BSA
IHC-P | WB | ICC/IF | |
---|---|---|---|
Human | Tested | Tested | Tested |
Mouse | Expected | Tested | Tested |
Rat | Expected | Tested | Tested |
Chicken | Expected | Expected | Expected |
Chinese hamster | Expected | Expected | Expected |
Cow | Expected | Expected | Expected |
Dog | Expected | Expected | Expected |
Drosophila melanogaster | Predicted | Predicted | Predicted |
Fish | Expected | Expected | Expected |
Guinea pig | Predicted | Predicted | Predicted |
Monkey | Predicted | Predicted | Predicted |
Pig | Predicted | Predicted | Predicted |
Rabbit | Expected | Expected | Expected |
Rhesus monkey | Predicted | Predicted | Predicted |
Sheep | Predicted | Predicted | Predicted |
Xenopus laevis | Expected | Expected | Expected |
Zebrafish | Predicted | Predicted | Predicted |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 0.20000-1.00000 µg/mL | Notes We recommend Goat Anti-Rabbit IgG H&L (Biotin) (Goat Anti-Rabbit IgG H&L (Biotin) ab6720) secondary antibody or Goat Anti-Rabbit IgG Fc (HRP) (Goat Anti-Rabbit IgG Fc (HRP) ab97200) secondary antibody. Perform heat-mediated antigen retrieval before commencing with IHC staining protocol. |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info - | Notes - |
Species Rat | Dilution info 0.20000-1.00000 µg/mL | Notes We recommend Goat Anti-Rabbit IgG H&L (Biotin) (Goat Anti-Rabbit IgG H&L (Biotin) ab6720) secondary antibody or Goat Anti-Rabbit IgG Fc (HRP) (Goat Anti-Rabbit IgG Fc (HRP) ab97200) secondary antibody. Perform heat-mediated antigen retrieval before commencing with IHC staining protocol. |
Species Chicken | Dilution info 0.2-1 µg/mL | Notes We recommend Goat Anti-Rabbit IgG H&L (Biotin) (Goat Anti-Rabbit IgG H&L (Biotin) ab6720) secondary antibod or Goat Anti-Rabbit IgG Fc (HRP) (Goat Anti-Rabbit IgG Fc (HRP) ab97200) secondary antibody. Perform heat-mediated antigen retrieval before commencing with IHC staining protocol. |
Species Chinese hamster | Dilution info 0.2-1 µg/mL | Notes We recommend Goat Anti-Rabbit IgG H&L (Biotin) (Goat Anti-Rabbit IgG H&L (Biotin) ab6720) secondary antibod or Goat Anti-Rabbit IgG Fc (HRP) (Goat Anti-Rabbit IgG Fc (HRP) ab97200) secondary antibody. Perform heat-mediated antigen retrieval before commencing with IHC staining protocol. |
Species Cow | Dilution info 0.2-1 µg/mL | Notes We recommend Goat Anti-Rabbit IgG H&L (Biotin) (Goat Anti-Rabbit IgG H&L (Biotin) ab6720) secondary antibod or Goat Anti-Rabbit IgG Fc (HRP) (Goat Anti-Rabbit IgG Fc (HRP) ab97200) secondary antibody. Perform heat-mediated antigen retrieval before commencing with IHC staining protocol. |
Species Dog | Dilution info 0.2-1 µg/mL | Notes We recommend Goat Anti-Rabbit IgG H&L (Biotin) (Goat Anti-Rabbit IgG H&L (Biotin) ab6720) secondary antibod or Goat Anti-Rabbit IgG Fc (HRP) (Goat Anti-Rabbit IgG Fc (HRP) ab97200) secondary antibody. Perform heat-mediated antigen retrieval before commencing with IHC staining protocol. |
Species Fish | Dilution info 0.2-1 µg/mL | Notes We recommend Goat Anti-Rabbit IgG H&L (Biotin) (Goat Anti-Rabbit IgG H&L (Biotin) ab6720) secondary antibod or Goat Anti-Rabbit IgG Fc (HRP) (Goat Anti-Rabbit IgG Fc (HRP) ab97200) secondary antibody. Perform heat-mediated antigen retrieval before commencing with IHC staining protocol. |
Species Rabbit | Dilution info 0.2-1 µg/mL | Notes We recommend Goat Anti-Rabbit IgG H&L (Biotin) (Goat Anti-Rabbit IgG H&L (Biotin) ab6720) secondary antibod or Goat Anti-Rabbit IgG Fc (HRP) (Goat Anti-Rabbit IgG Fc (HRP) ab97200) secondary antibody. Perform heat-mediated antigen retrieval before commencing with IHC staining protocol. |
Species Xenopus laevis | Dilution info 0.2-1 µg/mL | Notes We recommend Goat Anti-Rabbit IgG H&L (Biotin) (Goat Anti-Rabbit IgG H&L (Biotin) ab6720) secondary antibod or Goat Anti-Rabbit IgG Fc (HRP) (Goat Anti-Rabbit IgG Fc (HRP) ab97200) secondary antibody. Perform heat-mediated antigen retrieval before commencing with IHC staining protocol. |
Species | Dilution info | Notes |
---|---|---|
Species Guinea pig, Pig, Drosophila melanogaster, Monkey, Zebrafish, Rhesus monkey, Sheep | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 1/1000.00000 - 1/5000.00000 | Notes We recommend Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab6721) secondary antibody. A stronger signal is observed in chemiluminescent western blot when using 2% BSA as the blocking agent. Milk blocking is suitable for use with fluorescent detection systems. |
Species Mouse | Dilution info 1/1000.00000 - 1/5000.00000 | Notes We recommend Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab6721) secondary antibody. A stronger signal is observed in chemiluminescent western blot when using 2% BSA as the blocking agent. Milk blocking is suitable for use with fluorescent detection systems. |
Species Rat | Dilution info 1/1000.00000 - 1/5000.00000 | Notes We recommend Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab6721) secondary antibody. A stronger signal is observed in chemiluminescent western blot when using 2% BSA as the blocking agent. Milk blocking is suitable for use with fluorescent detection systems. |
Species | Dilution info | Notes |
---|---|---|
Species Chinese hamster | Dilution info 1/1000.00000 - 1/5000.00000 | Notes We recommend Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab6721) secondary antibody. A stronger signal is observed in chemiluminescent western blot when using 2% BSA as the blocking agent. Milk blocking is suitable for use with fluorescent detection systems. |
Species Fish | Dilution info 1/1000.00000 - 1/5000.00000 | Notes We recommend Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab6721) secondary antibody. A stronger signal is observed in chemiluminescent western blot when using 2% BSA as the blocking agent. Milk blocking is suitable for use with fluorescent detection systems. |
Species Xenopus laevis | Dilution info 1/1000.00000 - 1/5000.00000 | Notes We recommend Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab6721) secondary antibody. A stronger signal is observed in chemiluminescent western blot when using 2% BSA as the blocking agent. Milk blocking is suitable for use with fluorescent detection systems. |
Species Dog | Dilution info 1/1000.00000 - 1/5000.00000 | Notes We recommend Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab6721) secondary antibody. A stronger signal is observed in chemiluminescent western blot when using 2% BSA as the blocking agent. Milk blocking is suitable for use with fluorescent detection systems. |
Species Cow | Dilution info 1/1000.00000 - 1/5000.00000 | Notes We recommend Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab6721) secondary antibody. A stronger signal is observed in chemiluminescent western blot when using 2% BSA as the blocking agent. Milk blocking is suitable for use with fluorescent detection systems. |
Species Chicken | Dilution info 1/1000.00000 - 1/5000.00000 | Notes We recommend Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab6721) secondary antibody. A stronger signal is observed in chemiluminescent western blot when using 2% BSA as the blocking agent. Milk blocking is suitable for use with fluorescent detection systems. |
Species Rabbit | Dilution info 1/1000.00000 - 1/5000.00000 | Notes We recommend Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab6721) secondary antibody. A stronger signal is observed in chemiluminescent western blot when using 2% BSA as the blocking agent. Milk blocking is suitable for use with fluorescent detection systems. |
Species | Dilution info | Notes |
---|---|---|
Species Guinea pig, Pig, Drosophila melanogaster, Monkey, Zebrafish, Rhesus monkey, Sheep | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Rat | Dilution info 1 µg/mL | Notes - |
Species Human | Dilution info 1 µg/mL | Notes - |
Species Mouse | Dilution info 1 µg/mL | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Chicken | Dilution info 1 µg/mL | Notes - |
Species Chinese hamster | Dilution info 1 µg/mL | Notes - |
Species Cow | Dilution info 1 µg/mL | Notes - |
Species Dog | Dilution info 1 µg/mL | Notes - |
Species Fish | Dilution info 1 µg/mL | Notes - |
Species Rabbit | Dilution info 1 µg/mL | Notes - |
Species Xenopus laevis | Dilution info 1 µg/mL | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Guinea pig, Pig, Drosophila melanogaster, Monkey, Zebrafish, Rhesus monkey, Sheep | Dilution info - | Notes - |
Select an associated product type
Actin is a highly conserved protein that polymerizes to produce filaments that form cross-linked networks in the cytoplasm of cells (PubMed:25255767, PubMed:29581253). Actin exists in both monomeric (G-actin) and polymeric (F-actin) forms, both forms playing key functions, such as cell motility and contraction (PubMed:29581253). In addition to their role in the cytoplasmic cytoskeleton, G- and F-actin also localize in the nucleus, and regulate gene transcription and motility and repair of damaged DNA (PubMed:29925947). Part of the ACTR1A/ACTB filament around which the dynactin complex is built. The dynactin multiprotein complex activates the molecular motor dynein for ultra-processive transport along microtubules (By similarity).
Beta-actin, ACTB
Anti-beta Actin antibody is a rabbit polyclonal antibody that is used to detect beta Actin in ICC/IF, IHC-P, Western blot. Suitable for Chicken, Chinese hamster, Cow, Dog, Fish, Human, Mouse, Rabbit, Rat, Xenopus laevis samples.
- Cited in over 4,000 publications
- Trusted since 2003
pH: 7.4
Preservative: 0.02% Sodium azide
Constituents: 98.98% PBS, 1% BSA
From Jan 2024, QC testing of replenishment batches of this polyclonal changed. All tested and expected application and reactive species combinations are still covered by our Abcam product promise. However, we no longer test all applications. For more information on a specific batch, please contact our Scientific Support who will be happy to help. The immunogen used for this product shares 77% homology with Gamma actin/actin cytoplasmic 2. Cross-reactivity with this protein has not been confirmed experimentally.
Anti-beta Actin antibody (ab8227) is a rabbit polyclonal antibody and is validated for use in ICC/IF, IHC-P and WB.
Anti-beta Actin antibody (ab8227) was first used in a scientific publication in 1991 and has been cited over 4031 times in peer reviewed journals. It's performance in Western blot in human, mouse and rat samples is trusted by the scientific community.
Abcam's high quality validation processes ensure Anti-beta Actin antibody (ab8227) has high sensitivity and specificity.
Anti-beta Actin antibody (ab8227) has 104 independent reviews from customers.
beta Actin antibodies are often used as loading controls in Western Blot. Anti-beta Actin antibody has been verified in Western Blot samples and detects a band at 42kDa Molecular weight.
Anti-beta Actin antibody (ab8227) specifically detects beta Actin (UniProt ID: P60709; Molecular weight: 42kDa) and is sold in 50 µg selling sizes.
Beta-Actin (β-actin) is a highly conserved protein that plays a crucial role in cell motility, structure and integrity. It is one of the six actin isoforms found in humans and is a major component of the cytoskeleton in non-muscle cells. β-actin is involved in various cellular processes, including cell division, signaling and maintaining cell shape. Due to its ubiquitous expression and stability, β-actin is commonly used as a loading control in western blotting (WB) and other protein quantification assays. Understanding β-actin's functions and interactions is essential for studying cellular dynamics and various physiological processes.
Beta actin also known as beta cytoplasmic actin plays a central role in cell structure and motility. It is part of the actin protein family and is widely expressed in eukaryotic cells. The molecular weight of beta actin is approximately 42 kDa. It contributes to the formation of the cytoskeleton and participates in various cellular processes including movement and stability. Actin is abundant in all cell types providing structural integrity and flexibility.
Beta actin contributes to the maintenance of cell shape and is an important player in cell division and muscle contraction. It forms part of a larger actin filaments network often associating with other proteins to form the actin cytoskeleton complex. This complex supports cellular processes such as signaling intracellular trafficking and positioning of organelles. The dynamic polymerization and depolymerization of actin filaments are critical for cellular functions.
Beta actin functions in the regulation of important biological pathways such as the Rho/Rac/Cdc42 signaling pathway and the Wnt signaling pathway. These pathways are essential in numerous cellular activities including cell morphology and gene transcription. Beta actin closely interacts with proteins like myosin and tropomyosin which facilitate its role in muscle contraction and cell division and proteins such as Rac and Cdc42 which help govern cytoskeletal dynamics and cellular responses to extracellular stimuli.
Beta actin has links to cancers and muscular dystrophies. Aberrations in actin dynamics can result in tumor cell migration and metastasis making it a component of interest in cancer research. Additionally mutations or dysregulation in actin-associated proteins may contribute to muscular dystrophies affecting muscle function and strength. Beta actin's interactions with proteins like dystrophin involved in maintaining muscle integrity further highlight its relevance in both biological functions and disease contexts.
We have tested this species and application combination and it works. It is covered by our product promise.
We have not tested this specific species and application combination in-house, but expect it will work. It is covered by our product promise.
This species and application combination has not been tested, but we predict it will work based on strong homology. However, this combination is not covered by our product promise.
We do not recommend this combination. It is not covered by our product promise.
We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.
In the unlikely event of one of our products not working as expected, you are covered by our product promise.
Full details and terms and conditions can be found here:
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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
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
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
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
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
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
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
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
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
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
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
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
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
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).
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.
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).
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
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
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
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
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
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
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
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
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
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
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
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
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
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.
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.
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