Anti-Aryl hydrocarbon Receptor antibody [RPT9] - ChIP Grade
| Name | Anti-Aryl hydrocarbon Receptor antibody [RPT9] - ChIP Grade |
|---|---|
| Supplier | Abcam |
| Catalog | ab2769 |
| Prices | $395.00 |
| Sizes | 100 µl |
| Host | Mouse |
| Clonality | Monoclonal |
| Isotype | IgG |
| Clone | RPT9 |
| Applications | ICC/IF ICC/IF ELISA WB IHC-F ChIP IHC-P IP FC |
| Species Reactivities | Mouse, Rat, Human, Rabbit |
| Antigen | Recombinant fragment corresponding to Mouse Aryl hydrocarbon Receptor aa 12-31 |
| Description | Mouse Monoclonal |
| Gene | AHR |
| Conjugate | Unconjugated |
| Supplier Page | Shop |
Product images
Immunocytochemistry/Immunofluorescence analysis of Aryl Hydrocarbon Receptor shows staining in A2058 cells. Aryl Hydrocarbon Receptor (green), F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were incubated without (control) or with ab2769 (1:20) overnight at 4°C, washed with PBS and incubated with a DyLight-488 conjugated secondary antibody. Images were taken at 60X magnification.
Immunocytochemistry/Immunofluorescence analysis of Aryl Hydrocarbon Receptor shows staining in HeLa cells. Aryl Hydrocarbon Receptor (green), F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were incubated without (control) or with ab2769 (1:20) overnight at 4°C, washed with PBS and incubated with a DyLight-488 conjugated secondary antibody. Images were taken at 60X magnification.
Immunocytochemistry/Immunofluorescence analysis of Aryl Hydrocarbon Receptor shows staining in U251 cells. Aryl Hydrocarbon Receptor (green), F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were incubated without (control) or with ab2769 (1:20) overnight at 4°C, washed with PBS and incubated with a DyLight-488 conjugated secondary antibody. Images were taken at 60X magnification.
Western blot analysis of Aryl Hydrocarbon Receptor was performed by loading 40 ug of HEK293 lysate overexpressing Aryl Hydrocarbon Receptor (right lane) or empty vector control (left lane) and 10ul of a Prestained Protein Ladder onto a 4-20% Tris-HCl polyacrylamide gel. Proteins were transferred to a PVDF membrane and blocked with 5% BSA/TBST for at least 1 hour. The membrane was probed with an Aryl Hydrocarbon Receptor monoclonal antibody (ab2769) at a dilution of 1:1000 overnight at 4°C on a rocking platform then washed in TBS-0.1%Tween-20 and probed with a goat anti-mouse IgG-HRP secondary antibody at a dilution of 1:20,000 for 1 hour. Chemiluminescent detection was performed.
Flow cytometry analysis of Aryl Hydrocarbon Receptor showing positive staining in the nucleus and cytoplasm of NIH/3T3 cells compared to an isotype control (blue). Cells were harvested and adjusted to a concentration of 1-5x10^6 cells/ml. Cells were then fixed with 2% paraformaldehyde and washed with PBS. Cells were penetrated by dropping the supernatant and adding 90% methanol followed by incubation for 10 minutes at room temperature. Cells were blocked with a 2% solution of BSA-PBS for 30 min at room temperature and incubated with ab2769 at 1:50 for 60 min at room temperature. Cells were then incubated for 40 min at room temperature in the dark using a Dylight 488-conjugated goat anti-mouse IgG (H+L) secondary antibody and re-suspended in PBS for FACS analysis.
Flow cytometry analysis of Aryl Hydrocarbon Receptor showing positive staining in the nucleus and cytoplasm of PC-3 cells compared to an isotype control (blue). Cells were harvested and adjusted to a concentration of 1-5x10^6 cells/ml. Cells were then fixed with 2% paraformaldehyde and washed with PBS. Cells were penetrated by dropping the supernatant and adding 90% methanol followed by incubation for 10 minutes at room temperature. Cells were blocked with a 2% solution of BSA-PBS for 30 min at room temperature and incubated with ab2769 at 1:50 for 60 min at room temperature. Cells were then incubated for 40 min at room temperature in the dark using a Dylight 488-conjugated goat anti-mouse IgG (H+L) secondary antibody and re-suspended in PBS for FACS analysis.
Flow cytometry analysis of Aryl Hydrocarbon Receptor showing positive staining in the nucleus and cytoplasm of Hela cells compared to an isotype control (blue). Cells were harvested and adjusted to a concentration of 1-5x10^6 cells/ml. Cells were then fixed with 2% paraformaldehyde and washed with PBS. Cells were penetrated by dropping the supernatant and adding 90% methanol followed by incubation for 10 minutes at room temperature. Cells were blocked with a 2% solution of BSA-PBS for 30 min at room temperature and incubated with ab2769 at 1:50 for 60 min at room temperature. Cells were then incubated for 40 min at room temperature in the dark using a Dylight 488-conjugated goat anti-mouse IgG (H+L) secondary antibody and re-suspended in PBS for FACS analysis.
ICC/IF image of ab2769 stained HepG2 cells. The cells were 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 (ab2769, in a 1/200 dilution) 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.
This was a trial experiment to evaluate the association of AhR to the ABCG2 promoter using ab2769 at a 1/100 dilution for the IP (ChIP assay). Semiquantitative PCR was performed to evaluate the relative association of AhR with the proximal ABCG2 promoter in a S1 colon cancer cell line without treatment, or treated with depsipeptide (10 ng/mL 24h) or benzo(a)pyrene (10uM 24h). Cross-linking (X-ChIP) - 10 Mins 0 Secs Lane 1: DNA ladder (100 bp from promega) Inputs (lanes 2-4): S1 no treatment - serial dilution Inputs (lanes 5-7): S1 treated with depsipeptide - serial dilution Inputs (lanes 8-10): S1 treated with benzo(a)pyrene - serial dilution AhR ChIP (lane 11-13): use 1:1 diluted immunoprecipitate for PCR Lane 11=S1 no treatment lane 12=S1 depsipeptide 10ng/mL lane 13= S1 benzo(a)pyrene 10uM AhR ChIP (lane 14-16): use 1:2 diluted immunoprecipitate for PCR same order as lanes 11-13 Lane 17: H2O control for PCR.
![Overlay histogram showing HEK293 cells stained with ab2769 (red line). The cells were fixed with 100% 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 (ab2769, 1/20 dilution) for 30 min at 22°C. The secondary antibody used was a goat anti mouse-DyLight® 488 (IgG H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. 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 HEK293 cells fixed with 4% paraformaldehyde (10 min)/permeabilized in 0.1% PBS-Tween used under the same conditions.](http://www.bioprodhub.com/system/product_images/ab_products/2/sub_1/9990_Aryl-hydrocarbon-Receptor-Primary-antibodies-ab2769-14.jpg)
Overlay histogram showing HEK293 cells stained with ab2769 (red line). The cells were fixed with 100% 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 (ab2769, 1/20 dilution) for 30 min at 22°C. The secondary antibody used was a goat anti mouse-DyLight® 488 (IgG H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. 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 HEK293 cells fixed with 4% paraformaldehyde (10 min)/permeabilized in 0.1% PBS-Tween used under the same conditions.
Product References
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