Anti-beta Actin antibody
| Name | Anti-beta Actin antibody |
|---|---|
| Supplier | Abcam |
| Catalog | ab8227 |
| Prices | $404.00 |
| Sizes | 50 µg |
| Host | Rabbit |
| Clonality | Polyclonal |
| Isotype | IgG |
| Applications | IHC-F IP WB ICC/IF FC IHC-F IHC-P ICC/IF ICC/IF ELISA |
| Species Reactivities | Mouse, Rat, Sheep, Rabbit, Chicken, Guinea Pig, Bovine, Dog, Human, Pig, Xenopus, Drosophila, Fish, Monkey, Zebrafish, Monkey, Hamster |
| Antigen | Synthetic peptide derived from within residues 1 - 100 of Human beta Actin |
| Description | Rabbit Polyclonal |
| Gene | ACTB |
| Conjugate | Unconjugated |
| Supplier Page | Shop |
Product images
![IHC image of beta actin staining in a section of formalin-fixed paraffin-embedded [human normal 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 (ab6720, 1/1000 dilution) was used to detect the primary, and visualized using an HRP conjugated ABC system. Streptavidin HRP was used, 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](http://www.bioprodhub.com/system/product_images/ab_products/2/sub_1/13576_ab8227-242969-ab8227101ugmlab6720-2-1ugml-hu-colon-x20.jpg)
IHC image of beta actin staining in a section of formalin-fixed paraffin-embedded [human normal 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 (ab6720, 1/1000 dilution) was used to detect the primary, and visualized using an HRP conjugated ABC system. Streptavidin HRP was used, 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
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 (Ab97200, 1/200 dilution) was used for 1hr at room temperature. The section was counterstained with haematoxylin and mounted with DPX.*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre
Performed under reducing conditions.Exposure time : 1 minuteThis 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, and visualised using ECL development solution ab133406
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 ab175781 at a 1:10,000 dilution for 1hr at room temperature and then imaged using the Licor Odyssey CLx.
Rabbit polyclonal to beta actin (ab8227) at 1/5000. Lane 1: HeLa whole cell extract (20ug); lane 2: yeast cell extract (20ug); lane 3: mouse brain tissue lysate (20ug).Developed using Goat anti-rabbit HRP secondary antibody (ab6721) at 1/2000.
ICC/IF image of ab8227 stained human HeLa cells. The cells were methanol fixed (5 min) and incubated with the antibody (ab8227, 1µg/ml) for 1h at room temperature. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Image-iTTM FX Signal Enhancer was used as the primary blocking agent, 5% BSA was used for all other blocking steps. DAPI was used to stain the cell nuclei (blue).
IHC image of beta Actin staining in human liver FFPE section, performed on a BondTM system using the standard protocol F. 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 ab8227, 1µg/ml, for 8 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.
ab8227 staining beta Actin in human stomach tissue sections by Immunohistochemistry (frozen sections). Tissue was fixed with acetone and then blocked with 5% serum for 1 hour at 23°C followed by incubation with the primary antibody, at 1µg/ml, for 1 hour at 23°C. A diluted anti-rabbit HRP-conjugated goat polyclonal was used as secondary antibody.See Abreview
ab8227 used in Direct ELISA in NIH 3T3 murine fibroblasts. Primary antibody used at a 1/1000 dilution for 16 hours at 4°C. The secondary antibody used is an AP-conjugated Goat anti-rabbit used at a 1/2000 dilution. A blocking step was performed using 5% BSA for 1 hour at 23°C.See Abreview
ICC/IF image of ab8227 stained HeLa cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab8227, 5µg/ml) overnight at +4°C. The secondary antibody (green) was a goat anti-rabbit DyLight® 488 (IgG - H&L, pre-adsorbed) (ab96899) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
Exposure time : 30 secondsWestern blot using ab8227 at 1/1000.Lysates:Lane 1: HeLa cells (Human)Lane 2: 3T3 cells (Mouse)Lane 3: Fish LiverLane 4: Rabbit LiverLane 5: MDCK cells (Dog)Lane 6: EBTr cells (Cow)Lane 7: SL-29 cells (Chicken)Lane 8: CHO cells (Chinese Hamster)Lane 9: Xenopus embryoSecondary ab: anti-rabbit HRP (ab6721)Exposure time: 30 secLysates at 20µg/lane.Expected molecular weight: 41.7kDa.
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