H3K27me3 Antibody [BT164], Rabbit IgG

• Catalog Number: orb1671571
• Category: Antibodies
• Description: H3K27me3 Antibody [BT164], Rabbit IgG
• Clonality: Recombinant
• Clone Number: BT164
• Tested applications: ELISA, ICC, IF, IHC, IP, WB
• Reactivity: Human
• Isotype: IgG kappa
• Immunogen: BT164 is a mice-derrived lupus antibody that was established by selection on apoptotic chromatin.
• Concentration: batch dependent
• Conjugation: Unconjugated
• Storage: Store at 4°C for up to 3 months. For longer storage, aliquot and store at -20°C.
• Buffer/Preservatives: PBS with 0.02% Proclin 300.
• Alternative names: trimethyl-Histone H3 (Lys27), histone H3, H3K27me3
• Note: For research use only
• Expiration Date: 12 months from date of receipt.

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 r version of BT164 (orb1671571) at 10 ug/ml for 1h followed by Alexa Fluor® 488 secondary antibody (2 ug/ml)- showing membrane staining. The nuclear stain is DAPI (blue) and the actin stain is phalloidin (red). Panels show from left-right- top-bottom orb1671571- DAPI- merged channels and an isotype control. The isotype control was an unknown specificity antibody (3.0) followed by staining with Alexa Fluor® 488 secondary antibody.

Western Blot using anti-H3K27me3 antibody BT164 (Nuclear lysate of HeLa(A) (0.0003 ug/ml)- Jurkat(B) (0.0003 ug/ml) and K562(C) (0.01 ug/ml) (35 ug protein in RIPA buffer) were resolved on a SDS PAGE gel and blots were probed with the chimeric rsion of BT164 (orb1671571) before detection using an anti-rondary antibody. A primary incubation of 1h was used and protein was detected by chemiluminescence.

Flow cytometry using the anti-H3K27me3 antibody BT164 HeLa cells were fixed using 2% PFA and stained with anti-unknown specificity antibody (3.0; isotype control - black line) or the r1 version of BT164 (orb1671571 - blue line) at a dilution of 1:100 for 1h at RT. After washing- the bound antibody was detected using a goat anti-r AlexaFluor® 488 antibody at a dilution of 1:1000 and cells analyzed using a FACSCanto flow-cytometer.