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Histone H3 (Tri Methyl K27) Recombinant Antibody

The HeLa (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.). Cells stained with Primary Antibody for 30 min at room temperature,followed by secondary antibody incubation for 40 min at room temperature. Primary Antibody (green):Rabbit Anti-Histone H3 (Tri Methyl K27) antibody (bsm-60656R,1:100); Isotype Control (orange): Rabbit IgG (bs-0295P). Blank control (black): PBS. Acquisition of 20,000 events was performed.
The HeLa (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.). Cells stained with Primary Antibody for 30 min at room temperature,followed by secondary antibody incubation for 40 min at room temperature. Primary Antibody (green):Rabbit Anti-Histone H3 (Tri Methyl K27) antibody (bsm-60656R,1:100); Isotype Control (orange): Rabbit IgG (bs-0295P). Blank control (black): PBS. Acquisition of 20,000 events was performed.
Paraformaldehyde-fixed, paraffin embedded Mouse Liver; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with Histone H3 (Tri Methyl K27) Monoclonal Antibody, Unconjugated (bsm-60656R) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Mouse Liver; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with Histone H3 (Tri Methyl K27) Monoclonal Antibody, Unconjugated (bsm-60656R) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Human Liver; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with Histone H3 (Tri Methyl K27) Monoclonal Antibody, Unconjugated (bsm-60656R) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Human Liver; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with Histone H3 (Tri Methyl K27) Monoclonal Antibody, Unconjugated (bsm-60656R) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Mouse Testicles; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with Histone H3 (Tri Methyl K27) Monoclonal Antibody, Unconjugated (bsm-60656R) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Mouse Testicles; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with Histone H3 (Tri Methyl K27) Monoclonal Antibody, Unconjugated (bsm-60656R) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Human Testicles; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with Histone H3 (Tri Methyl K27) Monoclonal Antibody, Unconjugated (bsm-60656R) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Human Testicles; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with Histone H3 (Tri Methyl K27) Monoclonal Antibody, Unconjugated (bsm-60656R) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Mouse Colon; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with Histone H3 (Tri Methyl K27) Monoclonal Antibody, Unconjugated (bsm-60656R) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Mouse Colon; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with Histone H3 (Tri Methyl K27) Monoclonal Antibody, Unconjugated (bsm-60656R) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Human Colon; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with Histone H3 (Tri Methyl K27) Monoclonal Antibody, Unconjugated (bsm-60656R) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Human Colon; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with Histone H3 (Tri Methyl K27) Monoclonal Antibody, Unconjugated (bsm-60656R) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Human Colon Cancer; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with Histone H3 (Tri Methyl K27) Monoclonal Antibody, Unconjugated (bsm-60656R) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Human Colon Cancer; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with Histone H3 (Tri Methyl K27) Monoclonal Antibody, Unconjugated (bsm-60656R) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Mouse Kidney; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with Histone H3 (Tri Methyl K27) Monoclonal Antibody, Unconjugated (bsm-60656R) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Mouse Kidney; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with Histone H3 (Tri Methyl K27) Monoclonal Antibody, Unconjugated (bsm-60656R) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Human Glioma; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with Histone H3 (Tri Methyl K27) Monoclonal Antibody, Unconjugated (bsm-60656R) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Human Glioma; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with Histone H3 (Tri Methyl K27) Monoclonal Antibody, Unconjugated (bsm-60656R) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Human Cerebrum; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with Histone H3 (Tri Methyl K27) Monoclonal Antibody, Unconjugated (bsm-60656R) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Human Cerebrum; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with Histone H3 (Tri Methyl K27) Monoclonal Antibody, Unconjugated (bsm-60656R) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Human Pancreas; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with Histone H3 (Tri Methyl K27) Monoclonal Antibody, Unconjugated (bsm-60656R) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Human Pancreas; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with Histone H3 (Tri Methyl K27) Monoclonal Antibody, Unconjugated (bsm-60656R) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.
25 ug total protein per lane of various lysates (see on figure) probed with Histone H3 (Tri Methyl K27) monoclonal antibody, unconjugated (bsm-60656R) at 1:1000 dilution and 4\u00b0C overnight incubation. Followed by conjugated secondary antibody incubation at r.t. for 60 min.
25 ug total protein per lane of various lysates (see on figure) probed with Histone H3 (Tri Methyl K27) monoclonal antibody, unconjugated (bsm-60656R) at 1:1000 dilution and 4\u00b0C overnight incubation. Followed by conjugated secondary antibody incubation at r.t. for 60 min.
Chromatin immunoprecipitation was performed using cross-linked chromatin from 5\u00d710\u2076 HeLa cells per IP, with 5 \u03bcg of bsm-60656R antibody or 5 \u03bcg of normal rabbit IgG(negative control bs-0295P) , and 36 \u03bcL of Protein A\/G MagBeads per IP. The immunoprecipitated DNA was quantified by real-time PCR using primers specific for the human GAPDH-promoter, MyoD1 exon 1 and MyoD1-promoter regions. The data are presented as enrichment of each sample relative to the total amount of input at each amplicon.
Chromatin immunoprecipitation was performed using cross-linked chromatin from 5\u00d710\u2076 HeLa cells per IP, with 5 \u03bcg of bsm-60656R antibody or 5 \u03bcg of normal rabbit IgG(negative control bs-0295P) , and 36 \u03bcL of Protein A\/G MagBeads per IP. The immunoprecipitated DNA was quantified by real-time PCR using primers specific for the human GAPDH-promoter, MyoD1 exon 1 and MyoD1-promoter regions. The data are presented as enrichment of each sample relative to the total amount of input at each amplicon.
Paraformaldehyde-fixed, paraffin embedded Rat Lung; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with Histone H3 (Tri Methyl K27) Monoclonal Antibody, Unconjugated (bsm-60656R) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Rat Lung; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with Histone H3 (Tri Methyl K27) Monoclonal Antibody, Unconjugated (bsm-60656R) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Rat Testicles; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with Histone H3 (Tri Methyl K27) Monoclonal Antibody, Unconjugated (bsm-60656R) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Rat Testicles; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with Histone H3 (Tri Methyl K27) Monoclonal Antibody, Unconjugated (bsm-60656R) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Rat Colon; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with Histone H3 (Tri Methyl K27) Monoclonal Antibody, Unconjugated (bsm-60656R) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Rat Colon; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with Histone H3 (Tri Methyl K27) Monoclonal Antibody, Unconjugated (bsm-60656R) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Rat Spleen; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with Histone H3 (Tri Methyl K27) Monoclonal Antibody, Unconjugated (bsm-60656R) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Rat Spleen; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with Histone H3 (Tri Methyl K27) Monoclonal Antibody, Unconjugated (bsm-60656R) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Rat Thymus; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with Histone H3 (Tri Methyl K27) Monoclonal Antibody, Unconjugated (bsm-60656R) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Rat Thymus; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with Histone H3 (Tri Methyl K27) Monoclonal Antibody, Unconjugated (bsm-60656R) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.
Bioss unconjugated antibody vial

Applications

  • WB

Reactivity

  • Human
  • Mouse
  • Rat

Predicted Reactivity

  • Mouse
  • Rat
Overview
Catalog # bsm-60656r
Product Name Histone H3 (Tri Methyl K27) Recombinant Antibody
Applications
WB
Reactivity Human, Mouse, Rat
Predicted Reactivity Mouse, Rat
Specifications
Conjugation Unconjugated
Host Rabbit
Source KLH conjugated synthetic peptide derived from human Histone H3 (Tri Methyl K27)
Clonality Recombinant
Isotype IgG
Concentration Lot Dependent
Purification Purified by Protein A.
Storage Buffer 0.01M TBS (pH 7.4), 1% BSA, 0.02% Proclin 300, and 50% Glycerol
Storage Condition Shipped at 4C. Store at -20C for one year. Avoid repeated freeze/thaw cycles.
Target
Gene ID 8350
Swiss Prot P68431
Subcellular location Cell membrane
Synonyms H3K27me3; Tri-Methyl-Histone H3 (Lys27); Histone H3 (Tri-Methyl-Lys27); H3 histone family member E pseudogene; H3 histone family, member A; H3/A; H31_HUMAN; H3F3; H3FA; Hist1h3a; HIST1H3B; HIST1H3C; HIST1H3D; HIST1H3E; HIST1H3F; HIST1H3G; HIST1H3H; HIST1H3I; HIST1H3J; HIST3H3; histone 1, H3a; Histone cluster 1, H3a; Histone H3 3 pseudogene; Histone H3.1; Histone H3/a; Histone H3/b; Histone H3/c; Histone H3/d; Histone H3/f; Histone H3/h; Histone H3/i; Histone H3/j; Histone H3/k; Histone H3/l; H3.1; H3/d; H3C1; H3C10; H3C11; H3C12; H3C2; H3C3; H3C4; H3C7; H3C8; H3FD;
Application Dilution
WB 1:300-5000

Frequently asked questions

Which applications is Histone H3 (Tri Methyl K27) Recombinant Antibody (bsm-60656R) validated for, and at what dilution?

bsm-60656R is validated for WB (1:300-5000). These are recommended starting dilutions; optimal dilutions should be determined by the end user.

Which species does Histone H3 (Tri Methyl K27) Recombinant Antibody (bsm-60656R) react with?

Tested reactivity: Human, Mouse, Rat. Predicted reactivity (expected from sequence homology but not yet tested): Mouse, Rat.

How should bsm-60656R be stored?

Shipped at 4C. Store at -20C for one year. Avoid repeated freeze/thaw cycles. Storage buffer: 0.01M TBS (pH 7.4), 1% BSA, 0.02% Proclin 300, and 50% Glycerol

What immunogen was used to raise Histone H3 (Tri Methyl K27) Recombinant Antibody (bsm-60656R)?

KLH conjugated synthetic peptide derived from human Histone H3 (Tri Methyl K27)

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