Anti-HP1 gamma antibody - ChIP Grade
| Name | Anti-HP1 gamma antibody - ChIP Grade |
|---|---|
| Supplier | Abcam |
| Catalog | ab10480 |
| Prices | $398.00 |
| Sizes | 100 µg |
| Host | Rabbit |
| Clonality | Polyclonal |
| Isotype | IgG |
| Applications | WB IHC-P ICC/IF ICC/IF ChIPseq IP |
| Species Reactivities | Mouse, Human |
| Antigen | Synthetic peptide conjugated to KLH derived from within residues 1 - 100 of Mouse HP1 gamma |
| Blocking Peptide | Mouse HP1 gamma peptide |
| Description | Rabbit Polyclonal |
| Gene | CBX3 |
| Conjugate | Unconjugated |
| Supplier Page | Shop |
Product images
ab10480 staining HP1 gamma in Mouse 14.5dpc tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 1% BSA for 1 hour at room temperature; antigen retrieval was by heat mediation in Tris buffer pH 9. Samples were incubated with primary antibody (1/100 in 1% BSA + 1% FBS in TBS) for 16 hours. An undiluted HRP-conjugated Goat anti-rabbit IgG polyclonal was used as the secondary antibody.See Abreview
Immunofluorescent imaging of human cells (U2OS) with ab10480 reveals the expected exclusively nuclear staining, corresponding to the known nuclear localisation of HP1-gamma.IF was performed with a standard paraformaldehyde technique (fixed in PBS buffered PFH 4% for 5 minutes, permeabilised with 0.5% triton-PBS for 5 minutes, blocked with 5% milk / 0.2% tween for one hour. Primary antibody used at 1/200 in 5% milk / 0.2% TWEEN for one hour, secondary antibody for 30 minutes. All blocking and incubation steps carried out at 37 degrees C. Nuclei stained with Hoechst stain (blue). Immunofluorescent imaging of human cells (U2OS) with ab10480 reveals the expected exclusively nuclear staining, corresponding to the known nuclear localisation of HP1-gamma. IF was performed with a standard paraformaldehyde technique (fixed in PBS buffered PFH 4% for 5 minutes, permeabilised with 0.5% triton-PBS for 5 minutes, blocked with 5% milk / 0.2% tween for one hour.
ab10480 was used to stain HeLa cells in interphase in panel one. In the second panel, the cell is stained by ab10480 (green), DAPI (blue) and SH-CREST (red) which stains the centromeres. Fix 30 minutes on ice in 4% formaldehyde in PEM. Quench autofluorescence 2 x 5 minutes with 1 mg/ml Na borohydride or 100 mM ammonium chloride in PEM. Permeablize 30 minutes with 0.5% TX-100 in PEM. Block 30 minutes in 5% milk in TBST. Primary antibody incubated overnight at 4oC diluted 1/100 in 5% milk in TBST. Secondary antibody incubaged 1 hour at RT diluted in 5% milk in TBST. Post-fix 20 minutes on ice in 4% formaldehyde in PEM. Quench autofluorescence 2 x 5 minutes with ammonium chloride in PEM. Counterstain with DAPI in TBST. Mount with ProLong Gold antifade reagent from Invitrogen. Notes: Ample washing between each step. TBST = Tris buffered saline + 0.1% Tween. PEM = 80 mM K-PIPES, pH 6.8, 5 mM EGTA, 2 mM MgCl2.
ab10480 at a 1/200 dilution staining HP1 gamma in human skin epidermis tissue by Immunohistochemistry (Formalin fixed, paraffin embedded) incubated for 20 hours at 4°C. Heat mediated antigen retrieval step performed using citrate buffer pH 6.0, microwave 10 minutes. Permeabilized using Tris-Buffered Saline Tween-20. Blocked using 2% BSA for 1 hour at 22°C. Secondary used at a 1/200 dilution Goat polyclonal to Rabbit IgG - H&L (FITC) (ab6717). The nuclei are counterstained with propidium iodide (red).See Abreview
All lanes : Anti-HP1 gamma antibody - ChIP Grade (ab10480) at 1 µg/mlLane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell LysateLane 2 : MCF7 (Human breast adenocarcinoma cell line) Whole Cell LysateLane 3 : HEK293 (Human embryonic kidney cell line) Whole Cell LysateLysates/proteins at 10 µg per lane.SecondaryGoat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilutiondeveloped using the ECL techniquePerformed under reducing conditions.
developed using the ECL techniquePerformed under reducing conditions.
developed using the ECL techniquePerformed under reducing conditions.
![HP1 gamma was immunoprecipitated using 0.5mg Hek293 whole cell extract, 5µg of Rabbit polyclonal to HP1 gamma 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, Hek293 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 ab10480.Secondary: Mouse monoclonal [SB62a] anti-rabbit IgG light chain HRP (ab99697).Band: 24kDa; HP1 gamma](http://www.bioprodhub.com/system/product_images/ab_products/2/sub_3/4094_HP1-gamma-Primary-antibodies-ab10480-19.jpg)
HP1 gamma was immunoprecipitated using 0.5mg Hek293 whole cell extract, 5µg of Rabbit polyclonal to HP1 gamma 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, Hek293 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 ab10480.Secondary: Mouse monoclonal [SB62a] anti-rabbit IgG light chain HRP (ab99697).Band: 24kDa; HP1 gamma
Product References
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