Anti-hnRNP A1 antibody [9H10]
| Name | Anti-hnRNP A1 antibody [9H10] |
|---|---|
| Supplier | Abcam |
| Catalog | ab5832 |
| Prices | $404.00 |
| Sizes | 100 µl |
| Host | Mouse |
| Clonality | Monoclonal |
| Isotype | IgG2b |
| Clone | 9H10 |
| Applications | FC IHC-P ELISA WB IP ICC/IF ICC/IF |
| Species Reactivities | Mouse, Human |
| Antigen | Full length hnRNPA1 native protein (partially purified) from HeLa cells (Human) |
| Description | Mouse Monoclonal |
| Gene | HNRNPA1 |
| Conjugate | Unconjugated |
| Supplier Page | Shop |
Product images
ab5832 at 1/2000 staining GM10725 cells (Lymphoblastoid cell line). The cells were paraformaldehyde fixed and blocked with BSA prior to incubation with the antibody for 12 hours. An Alexa-Fluor ® 594 conjugated donkey anti-mouse antibody was used as the secondary. Cells were mounted in DAPI and observed using a confocal microscope.See Abreview
Ab5832 staining human lung. Staining is localized to the nucleus.Left panel: with primary antibody at 1 ug/ml. Right panel: isotype control.Sections were stained using an automated system (Dako PT Link), at room temperature. Sections were rehydrated and antigen retrieved with the Dako 3-in-1 antigen retrieval buffer, citrate pH 6.0. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 minutes. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 minutes 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 we recommend to optimize the primary antibody concentration and incubation time (overnight incubation), and amplification may be required.
![All lanes : Anti-hnRNP A1 antibody [9H10] (ab5832) at 1 µg/mlLane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell LysateLane 2 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell LysateLane 3 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell LysateLane 4 : HEK293 (Human embryonic kidney cell line) Whole Cell LysateLysates/proteins at 10 µg per lane.SecondaryGoat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilutiondeveloped using the ECL techniquePerformed under reducing conditions.](http://www.bioprodhub.com/system/product_images/ab_products/2/sub_3/3252_hnRNP-A1-Primary-antibodies-ab5832-8.jpg)
All lanes : Anti-hnRNP A1 antibody [9H10] (ab5832) at 1 µg/mlLane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell LysateLane 2 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell LysateLane 3 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell LysateLane 4 : HEK293 (Human embryonic kidney cell line) Whole Cell LysateLysates/proteins at 10 µg per lane.SecondaryGoat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilutiondeveloped using the ECL techniquePerformed under reducing conditions.
![All lanes : Anti-hnRNP A1 antibody [9H10] (ab5832) at 1/1000 dilutionLane 1 : Mouse NSC34 whole cell lysate with control siRNALane 2 : Mouse NSC34 whole cell lysate with hnRNP A1 siRNALysates/proteins at 10 µg per lane.SecondaryIRDye® 700DX-conjugated Donkey anti-Mouse IgG at 1/3000 dilutionPerformed under reducing conditions.](http://www.bioprodhub.com/system/product_images/ab_products/2/sub_3/3253_hnRNP-A1-Primary-antibodies-ab5832-7.jpg)
All lanes : Anti-hnRNP A1 antibody [9H10] (ab5832) at 1/1000 dilutionLane 1 : Mouse NSC34 whole cell lysate with control siRNALane 2 : Mouse NSC34 whole cell lysate with hnRNP A1 siRNALysates/proteins at 10 µg per lane.SecondaryIRDye® 700DX-conjugated Donkey anti-Mouse IgG at 1/3000 dilutionPerformed under reducing conditions.
![Overlay histogram showing Jurkat cells stained with ab5832 (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 (ab5832, 1µg/1x106 cells) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG2b [PLPV219] (ab91366, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in Jurkat cells fixed with 4% paraformaldehyde/permeabilized in 0.1% PBS-Tween used under the same conditions.](http://www.bioprodhub.com/system/product_images/ab_products/2/sub_3/3254_hnRNP-A1-Primary-antibodies-ab5832-9.jpg)
Overlay histogram showing Jurkat cells stained with ab5832 (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 (ab5832, 1µg/1x106 cells) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG2b [PLPV219] (ab91366, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in Jurkat cells fixed with 4% paraformaldehyde/permeabilized in 0.1% PBS-Tween used under the same conditions.
hnRNP A1 was immunoprecipitated using 0.5mg Hela whole cell extract, 5µg of Mouse monoclonal to hnRNP A1 (ab5832) and 50µl of protein G magnetic beads (+). No antibody was added to the control (-). The antibody was incubated under agitation with Protein G beads for 10min, Hela whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab5832.Secondary: Goat polyclonal to mouse IgG light chain specific (HRP) at 1/5000 dilution.Band: 37kDa: hnRNP A1; 42kDa:We are unsure as to the identity of this extra band.
Product References
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HnRNP A1 controls a splicing regulatory circuit promoting - HnRNP A1 controls a splicing regulatory circuit promoting
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