Anti-PKR (phospho T446) antibody [E120] - ChIP Grade

• Supplier: Abcam
• Catalog: ab32036
• Prices: $394.00
• Sizes: 100 µl
• Host: Rabbit
• Clonality: Monoclonal
• Isotype: IgG
• Clone: E120
• Applications: ChIP FC IHC-P WB ICC/IF ICC/IF IP
• Species Reactivities: Human, Mouse
• Antigen: A synthetic phospho-peptide corresponding to residues surrounding Thr446 of human PKR (Interferon-inducible RNA-dependent protein kinase)
• Blocking Peptide: PKR (phospho T446) peptide
• Description: Rabbit Monoclonal
• Gene: EIF2AK2
• Conjugate: Unconjugated

Product images

All lanes : Anti-Estrogen Receptor alpha antibody [E115] - ChIP Grade (ab32063) at 1/1000 dilution (purified)Lane 1 : T47-D cell lysateLane 2 : MCF-7 cell lysateLysates/proteins at 20 µg per lane.SecondaryHRP goat ant-rabbit (H+L) at 1/1000 dilution

Immunohistochemical staining of paraffin embedded human endometrial adenocarcinoma with purified ab32036 at a dilution of 1/200. A HRP goat anti-rabbit (ab97051) was used as the secondary antibody at a dilution of 1/500 and the sample was counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0.

Immunofluorescent staining of MCF-7 cells (fixed in 4% PFA, permeabilized with 0.1% Triton X 100) using purified ab32036 at a dilution of 1/200. An Alexa Fluor® 647 goat anti-rabbit antibody was used as the secondary at a dilution of 1/500 and the cells were counter stained with DAPI.

Chromatin was prepared from MCF-7+β-estraiol 30min, and MCF-7 cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10 min. The ChIP was performed with 25 μg of chromatin, 4 μg of purified ab32063 (blue), and 20 μL of anti-rabbit IgG sepharose beads. Rabbit normal IgG was added to the beads control (yellow). The immunoprecipitated DNA was quantified by real time PCR (SYBR approach). Primers are located in the 2nd intron of TFF1 gene.MCF7 Cells were treated as below:MCF-7 starved overnight, then 10nM β-Estradiol together treat in 2%FBS media for 30min.Control MCF-7 was starved overnight, then in 2% FBS media for 30mins.Primer information:Located to the 2 intron of TFF1 gene.Sequence:Forward: 5' -agtctcctccaacctgacctt-3'Reverse: 5' -ttccggccatctctcactat-3'

Overlay histogram showing HeLa cells stained with unpurified ab32036 (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 (ab32036, 1/1000 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.

All lanes : Anti-PKR (phospho T446) antibody [E120] - ChIP Grade (ab32036) at 1/1000 dilution (unpurified)Lane 1 : Hela cell lysateLane 2 : Hela cell lysate + IFN + CalyculinALane 3 : Hela cell lysate + Pptase

Immunohistochemical analysis of PKR (phospho T446) expression in normal colon tissue, using 1/50 unpurified ab32036.

Product References

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