H3 Rabbit Monoclonal Antibody [Clone ID: BT164]

CAT#: TA386429

Recombinant Anti-H3K27me3 (Clone BT164)

Product Images

Flow cytometry using the anti-H3K27me3 antibody BT164 (TA386429). HeLa cells were fixed using 2% PFA and stained with anti-unknown specificity antibody (TA385792; isotype control, black line) or the rabbit IgG1 version of BT164 (TA386429, 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.

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  Western Blot using anti-H3K27me3 antibody BT164 (TA386429) Nuclear lysate of HeLa(A) (0.0003µg/ml), Jurkat(B) (0.0003µg/ml) and K562(C) (0.01ug/ml) (35µg protein in RIPA buffer) were resolved on a SDS PAGE gel and blots were probed with the chimeric rabbit version of BT164 (TA386429) before detection using an anti-rabbit secondary antibody. A primary incubation of 1h was used and protein was detected by chemiluminescence.

Specifications

Product Data

• Clone Name: BT164
• Applications: ELISA, ICC, IF, IHC, IP, WB
• Reactivities: Human
• Host: Rabbit
• Isotype: IgG, kappa
• Clonality: Monoclonal
• Immunogen: BT164 is a mice-derrived lupus antibody that was established by selection on apoptotic chromatin.
• Specificity: The epitope of BT164 was mapped in the N-terminal tail of histone H3 (27-KSAPAT-32) and included the apoptosis-induced trimethylation of K27.
  
  BT164 recognizes an apoptosis-induced trimethylation of K27 on N-terminal tail of histone H3 which is a target for autoantibodies in patients and mice with SLE (systemic lupus erythematosus) (van Bavel et al., 2011; pmid: 20699234). Consequently, this antibody is recommended for the detection and studying of SLE. ELISA studies demonstrated that BT164 exhibits preferential activity with the apoptotic nucleosomes in comparison with the control nucleosomes (van Bavel et al., 2011; pmid: 20699234). It can be used for the detection of apoptosis in general. Furthermore, this antibody was demonstrated to label cells with methylated histone-3 on K27 via immunofluorescence and it also immunoprecipitated chromatin with such a methylation profile, further demonstrating its ability to specifically recognize markers characteristic of SLE (van Bavel et al., 2011; pmid: 20699234).
• Formulation: PBS with 0.02% Proclin 300.
• Concentration: lot specific
• Conjugation: Unconjugated
• Storage: Please store at 4°C for up to 3 months. For longer storage, aliquot and store at -20°C. Avoid freeze and thaw cycles.
• Stability: 3 years from dispatch.
• Note: This chimeric rabbit antibody was made using the variable domain sequences of the original Mouse IgG2b format, for improved compatibility with existing reagents, assays and techniques.