Anti-Actin antibody [ACTN05 (C4)]
| Name | Anti-Actin antibody [ACTN05 (C4)] |
|---|---|
| Supplier | Abcam |
| Catalog | ab3280 |
| Prices | $404.00 |
| Sizes | 500 µl |
| Host | Mouse |
| Clonality | Monoclonal |
| Isotype | IgG1 |
| Clone | ACTN05 (C4) |
| Applications | FC IP WB IHC-F IHC-P ICC/IF ICC/IF |
| Species Reactivities | Mouse, Rat, Rabbit, Bovine, Dog, Human, Pig, Dictyostelium discoideum, Chicken |
| Antigen | Chicken gizzard actin |
| Description | Mouse Monoclonal |
| Gene | ACTA1 |
| Conjugate | Unconjugated |
| Supplier Page | Shop |
Product images
![All lanes : Anti-Actin antibody [ACTN05 (C4)] (ab3280)Lane 1 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell LysateLane 2 : MDA-MB-231 (Human breast adenocarcinoma cell line) Whole Cell Lysate Whole Cell LysateLane 3 : HeLa (Human epithelial carcinoma cell line) Whole Cell LysateLane 4 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell LysateLane 5 : MDA-MB-231 (Human breast adenocarcinoma cell line) Whole Cell Lysate Whole Cell LysateLane 6 : HeLa (Human epithelial carcinoma cell line) Whole Cell LysateLane 7 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell LysateLane 8 : MDA-MB-231 (Human breast adenocarcinoma cell line) Whole Cell Lysate Whole Cell LysateLane 9 : HeLa (Human epithelial carcinoma cell line) Whole Cell LysateLysates/proteins at 10 µg per lane.developed using the ECL techniquePerformed under reducing conditions.](http://www.bioprodhub.com/system/product_images/ab_products/2/sub_1/2146_Actin-Primary-antibodies-ab3280-1.jpg)
All lanes : Anti-Actin antibody [ACTN05 (C4)] (ab3280)Lane 1 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell LysateLane 2 : MDA-MB-231 (Human breast adenocarcinoma cell line) Whole Cell Lysate Whole Cell LysateLane 3 : HeLa (Human epithelial carcinoma cell line) Whole Cell LysateLane 4 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell LysateLane 5 : MDA-MB-231 (Human breast adenocarcinoma cell line) Whole Cell Lysate Whole Cell LysateLane 6 : HeLa (Human epithelial carcinoma cell line) Whole Cell LysateLane 7 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell LysateLane 8 : MDA-MB-231 (Human breast adenocarcinoma cell line) Whole Cell Lysate Whole Cell LysateLane 9 : HeLa (Human epithelial carcinoma cell line) Whole Cell LysateLysates/proteins at 10 µg per lane.developed using the ECL techniquePerformed under reducing conditions.
![All lanes : Anti-Actin antibody [ACTN05 (C4)] (ab3280) at 1/2000 dilutionLane 1 : Mouse liver whole tissue lysate (mouse 1).Lane 2 : Mouse liver whole tissue lysate (mouse 2).Lane 3 : Mouse liver whole tissue lysate (mouse 3).Lysates/proteins at 50 µg per lane.SecondaryAn HRP-conjugated goat anti-mouse IgG polyclonal. at 1/5000 dilutiondeveloped using the ECL techniquePerformed under reducing conditions.](http://www.bioprodhub.com/system/product_images/ab_products/2/sub_1/2147_Actin-Primary-antibodies-ab3280-4.jpg)
All lanes : Anti-Actin antibody [ACTN05 (C4)] (ab3280) at 1/2000 dilutionLane 1 : Mouse liver whole tissue lysate (mouse 1).Lane 2 : Mouse liver whole tissue lysate (mouse 2).Lane 3 : Mouse liver whole tissue lysate (mouse 3).Lysates/proteins at 50 µg per lane.SecondaryAn HRP-conjugated goat anti-mouse IgG polyclonal. at 1/5000 dilutiondeveloped using the ECL techniquePerformed under reducing conditions.
ab3280 staining Actin in mouse heart tissue sections by IHC-Fr (formaldehyde-fixed frozen sections). Tissue samples were fixed with formaldehyde, permeabilized with 0.1% Triton X-100 and blocked with 5% serum for 1 hour at 25°C. The sample was incubated with primary antibody (1/250 in PBS-Tween) at 4°C for 12 hours. An Alexa Fluor®488-conjugated Goat polyclonal to mouse IgG (1/250) was used as secondary antibody. Nuclei were stained with DAPI.See Abreview
ab3280 (1µg/ml) staining actin in human colon using an automated system (DAKO Autostainer Plus). Using this protocol there is strong cytoplasmic staining throughout the tissue.Sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffer citrate pH6.1 in a DAKO PT link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 mins. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 min and detected with Dako envision flex amplification kit for 30 minutes. Colorimetric detection was completed with Diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that, for manual staining, optimization of primary antibody concentration and incubation time is recommended. Signal amplification may be required.
ICC/IF image of ab3280 stained Hepp cells. The cells were 4% formaldehyde fixed (10 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 (ab3280, 10µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse IgG (H+L) used at a 1/1000 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.
ICC/IF image of ab3280 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 (ab3280, 5µg/ml) overnight at +4°C. The secondary antibody (green) was a goat anti-mouse DyLight® 488 (IgG - H&L, pre-adsorbed) (ab96879) 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.
![Overlay histogram showing HeLa cells stained with ab3280 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab3280, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was a goat anti-mouse DyLight® 488 (IgG; H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HeLa cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.](http://www.bioprodhub.com/system/product_images/ab_products/2/sub_1/2152_Actin-Primary-antibodies-ab3280-85.jpg)
Overlay histogram showing HeLa cells stained with ab3280 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab3280, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was a goat anti-mouse DyLight® 488 (IgG; H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HeLa cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.
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