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Di-Methyl-Histone H3 (K5) Monoclonal Antibody

4% Paraformaldehyde-fixed HeLa (treated with Trichostatin A (TSA)(1\u03bcM,18h) ) (H) cell; Triton X-100 at r.t. for 20 min; Antibody incubation with (Di-Methyl-Histone H3 (K5)) monoclonal Antibody, unconjugated (bsm-63100R) 1:100, 90 min at 37\u00b0C; followed by BF488 conjugated Goat Anti-Rabbit IgG antibody (green, bs-60295G-BF488) at 37\u00b0C for 90 min, DAPI (blue, C02-04002) was used to stain the cell nuclei. PBS instead of the primary antibody was used as the blank control.
4% Paraformaldehyde-fixed HeLa (treated with Trichostatin A (TSA)(1\u03bcM,18h) ) (H) cell; Triton X-100 at r.t. for 20 min; Antibody incubation with (Di-Methyl-Histone H3 (K5)) monoclonal Antibody, unconjugated (bsm-63100R) 1:100, 90 min at 37\u00b0C; followed by BF488 conjugated Goat Anti-Rabbit IgG antibody (green, bs-60295G-BF488) at 37\u00b0C for 90 min, DAPI (blue, C02-04002) was used to stain the cell nuclei. PBS instead of the primary antibody was used as the blank control.
The HeLa (treated with Trichostatin A (TSA)(1\u03bcM, 18h)) (H) cells were fixed with 4% PFA (10 min at r.t.) and then permeabilized with 90% ice-cold methanol for 20 min at -20\u2103,the cells then were incubated in 5%BSA to block non-specific protein-protein interactions (30 min at r.t.), followed by secondary antibody incubation for 40 min at room temperature. Primary Antibody (green):Rabbit Anti-Di-Methyl-Histone H3 (K5) antibody (bsm-63100R,1:100); Isotype Control (orange): Rabbit IgG (bs-0295P). Blank control (black): PBS. Acquisition of 20,000 events was performed.\r\n
The HeLa (treated with Trichostatin A (TSA)(1\u03bcM, 18h)) (H) cells were fixed with 4% PFA (10 min at r.t.) and then permeabilized with 90% ice-cold methanol for 20 min at -20\u2103,the cells then were incubated in 5%BSA to block non-specific protein-protein interactions (30 min at r.t.), followed by secondary antibody incubation for 40 min at room temperature. Primary Antibody (green):Rabbit Anti-Di-Methyl-Histone H3 (K5) antibody (bsm-63100R,1:100); Isotype Control (orange): Rabbit IgG (bs-0295P). Blank control (black): PBS. Acquisition of 20,000 events was performed.\r\n
HeLa (H) treated with Trichostatin A (TSA) (1\u03bcM) for 18 h, 25 \u03bcg total protein per lane of cell lysates (see on figure) probed with Di-Methyl-Histone H3 (K5) monoclonal antibody, unconjugated (bsm-63100R) at 1:1000 dilution and 4\u00b0C overnight incubation. Followed by conjugated secondary antibody incubation at r.t. for 60 min.
HeLa (H) treated with Trichostatin A (TSA) (1\u03bcM) for 18 h, 25 \u03bcg total protein per lane of cell lysates (see on figure) probed with Di-Methyl-Histone H3 (K5) monoclonal antibody, unconjugated (bsm-63100R) at 1:1000 dilution and 4\u00b0C overnight incubation. Followed by conjugated secondary antibody incubation at r.t. for 60 min.
Bioss unconjugated antibody vial

Applications

  • WB
  • IHC-P
  • IHC-F
  • IP
  • IF
  • ChIP
  • Flow-Cyt
  • ICC/IF

Reactivity

  • Human
  • Mouse
  • Rat
Overview
Catalog # bsm-63100r-100uL
Product Name Di-Methyl-Histone H3 (K5) Monoclonal Antibody
Applications
WB IHC-P IHC-F IP IF CHIP FLOW-CYT ICC/IF
Reactivity Human, Mouse, Rat
Specifications
Conjugation Unconjugated
Host Rabbit
Source A synthesized peptide derived from human Histone H3.1 around the methylation site of K5
Clonality Recombinant
Clone # 3F8
Isotype IgG
Concentration Lot dependent
Purification Purified by Protein A.
Storage Buffer 10mM phosphate buffered saline(pH 7.4) with 150mM sodium chloride, 0.05% BSA, 0.02% Proclin300 and 50% glycerol.
Storage Condition Shipped at 4C. Store at -20C for one year. Avoid repeated freeze/thaw cycles.
Target
Swiss Prot P68431
Subcellular location Nucleus
Synonyms H3C1 | Histone H3 (me2-K5); me2-Histone H3; di-methyl-Histone H3; H3K5me2; me2-H3;
Application Dilution
WB =1:1000-2000
IHC-P IHC-P=1:100-200
IHC-F IHC-F=1:100-200
IP IP=1:20-50
IF IF=1:50-200
ChIP ChIP=1:20-50
Flow-Cyt Flow-Cyt=1:50-100
ICC/IF ICC/IF=1:100-500

Frequently asked questions

Which applications is Di-Methyl-Histone H3 (K5) Monoclonal Antibody (bsm-63100R) validated for, and at what dilution?

bsm-63100R is validated for WB (WB=1:1000-2000); IHC-P (IHC-P=1:100-200); IHC-F (IHC-F=1:100-200); IP (IP=1:20-50); IF (IF=1:50-200); ChIP (ChIP=1:20-50); Flow-Cyt (Flow-Cyt=1:50-100); ICC/IF (ICC/IF=1:100-500). These are recommended starting dilutions; optimal dilutions should be determined by the end user.

Which species does Di-Methyl-Histone H3 (K5) Monoclonal Antibody (bsm-63100R) react with?

Tested reactivity: Human, Mouse, Rat.

How should bsm-63100R be stored?

Shipped at 4C. Store at -20C for one year. Avoid repeated freeze/thaw cycles. Storage buffer: 10mM phosphate buffered saline(pH 7.4) with 150mM sodium chloride, 0.05% BSA, 0.02% Proclin300 and 50% glycerol.

What immunogen was used to raise Di-Methyl-Histone H3 (K5) Monoclonal Antibody (bsm-63100R)?

A synthesized peptide derived from human Histone H3.1 around the methylation site of K5

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