Anti-H3K27me3 [BT164]

• Catalog Number: orb758980
• Category: Antibodies
• Description: Mouse monoclonal antibody to H3K27me3
• Species/Host: Mouse
• Clonality: Monoclonal
• Clone Number: BT164
• Tested applications: ELISA, ICC, IF, IHC, IP, WB
• Reactivity: Human
• Isotype: IgG2b
• Immunogen: BT164 is a mice-derrived lupus antibody that was established by selection on apoptotic chromatin.
• Concentration: 1 mg/ml
• Purity: Purified
• Conjugation: Unconjugated
• Target: H3K27me3
• Storage: Maintain refrigerated at 2-8°C for up to 2 weeks. For long term storage store at -20°C in small aliquots to prevent freeze-thaw cycles.
• Buffer/Preservatives: PBS with 0.02% Proclin 300.
• Alternative names: trimethyl-Histone H3 (Lys27); histone H3; H3K27; T
• Note: For research use only
• Expiration Date: 12 months from date of receipt.

Flow cytometry using the anti-H3K27me3 antibody BT164. HeLa cells were fixed using 2% PFA and stained with anti-unknown specificity antibody (orb256458; isotype control, black line) or the rabbit IgG1 version of BT164 (orb758979, blue line) at a dilution of 1:100 for 1h at RT. After washing, the bound antibody was detected using a goat anti-rabbit IgG AlexaFluor® 488 antibody at a dilution of 1:1000 and cells analyzed using a FACSCanto flow-cytometer.

Immunofluorescence staining of fixed HeLa cells with anti-H3K27me3 antibody BT164. Immunofluorescence analysis of paraformaldehyde fixed HeLa cells on Shi-fix™ coverslips stained with the chimeric rabbit IgG version of BT164 (orb758979) at 10 µg/ml for 1h followed by Alexa Fluor® 488 secondary antibody (2 µg/ml), showing membrane staining. The nuclear stain is DAPI (blue) and the actin stain is phalloidin (red). Panels show from left-right, top-bottom orb758979, DAPI, merged channels and an isotype control. The isotype control was an unknown specificity antibody (orb256458) followed by staining with Alexa Fluor® 488 secondary antibody.

Western Blot using anti-H3K27me3 antibody BT164 Nuclear lysate of HeLa(A) (0.0003 µg/ml), Jurkat(B) (0.0003 µg/ml) and K562(C) (0.01 ug/ml) (35 µg protein in RIPA buffer) were resolved on a SDS PAGE gel and blots were probed with the chimeric rabbit version of BT164 (orb758979) before detection using an anti-rabbit secondary antibody. A primary incubation of 1h was used and protein was detected by chemiluminescence.