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Histone H3.3(S22) Monoclonal Antibody

4% Paraformaldehyde-fixed HeLa (treated with 100ng\/ml of nocodazole for 18 h) (H) cell; Triton X-100 at r.t. for 20 min; Antibody incubation with (phospho-Histone H3.3(S22)) monoclonal Antibody, unconjugated (bsm-63099R) 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 100ng\/ml of nocodazole for 18 h) (H) cell; Triton X-100 at r.t. for 20 min; Antibody incubation with (phospho-Histone H3.3(S22)) monoclonal Antibody, unconjugated (bsm-63099R) 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.
HeLa (H) cells were treated with or without Calyculin A (100nM) for 30 min, 25 \u03bcg total protein per lane of cell lysates (see on figure) probed with phospho-Histone H3.3(S22) monoclonal antibody, unconjugated (bsm-63099R) at 1:1000 dilution and 4\u00b0C overnight incubation. Followed by conjugated secondary antibody incubation at r.t. for 60 min.
HeLa (H) cells were treated with or without Calyculin A (100nM) for 30 min, 25 \u03bcg total protein per lane of cell lysates (see on figure) probed with phospho-Histone H3.3(S22) monoclonal antibody, unconjugated (bsm-63099R) at 1:1000 dilution and 4\u00b0C overnight incubation. Followed by conjugated secondary antibody incubation at r.t. for 60 min.
Paraformaldehyde-fixed, paraffin embedded Mouse Testicles; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with phospho-Histone H3.3(S22) Monoclonal Antibody, Unconjugated (bsm-63099R) 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 phospho-Histone H3.3(S22) Monoclonal Antibody, Unconjugated (bsm-63099R) 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 phospho-Histone H3.3(S22) Monoclonal Antibody, Unconjugated (bsm-63099R) 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 phospho-Histone H3.3(S22) Monoclonal Antibody, Unconjugated (bsm-63099R) 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 phospho-Histone H3.3(S22) Monoclonal Antibody, Unconjugated (bsm-63099R) 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 phospho-Histone H3.3(S22) Monoclonal Antibody, Unconjugated (bsm-63099R) 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 phospho-Histone H3.3(S22) Monoclonal Antibody, Unconjugated (bsm-63099R) 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 phospho-Histone H3.3(S22) Monoclonal Antibody, Unconjugated (bsm-63099R) 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 phospho-Histone H3.3(S22) Monoclonal Antibody, Unconjugated (bsm-63099R) 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 phospho-Histone H3.3(S22) Monoclonal Antibody, Unconjugated (bsm-63099R) 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 Skin; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with phospho-Histone H3.3(S22) Monoclonal Antibody, Unconjugated (bsm-63099R) 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 Skin; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with phospho-Histone H3.3(S22) Monoclonal Antibody, Unconjugated (bsm-63099R) 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 Prostate Cancer; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with phospho-Histone H3.3(S22) Monoclonal Antibody, Unconjugated (bsm-63099R) 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 Prostate Cancer; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with phospho-Histone H3.3(S22) Monoclonal Antibody, Unconjugated (bsm-63099R) 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 Breast Cancer; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with phospho-Histone H3.3(S22) Monoclonal Antibody, Unconjugated (bsm-63099R) 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 Breast Cancer; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with phospho-Histone H3.3(S22) Monoclonal Antibody, Unconjugated (bsm-63099R) 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 Glioblastoma; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with phospho-Histone H3.3(S22) Monoclonal Antibody, Unconjugated (bsm-63099R) 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 Glioblastoma; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with phospho-Histone H3.3(S22) Monoclonal Antibody, Unconjugated (bsm-63099R) 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 Small Intestine; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with phospho-Histone H3.3(S22) Monoclonal Antibody, Unconjugated (bsm-63099R) 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 Small Intestine; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with phospho-Histone H3.3(S22) Monoclonal Antibody, Unconjugated (bsm-63099R) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.
The HeLa (treated with 100ng\/ml of nocodazole for 18 h) (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-phospho-Histone H3.3(S22) antibody (bsm-63099R,1:100); Isotype Control (orange): Rabbit IgG (bs-0295P). Blank control (black): PBS. Acquisition of 20,000 events was performed.
The HeLa (treated with 100ng\/ml of nocodazole for 18 h) (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-phospho-Histone H3.3(S22) antibody (bsm-63099R,1:100); Isotype Control (orange): Rabbit IgG (bs-0295P). Blank control (black): PBS. Acquisition of 20,000 events was performed.
Paraformaldehyde-fixed, paraffin embedded Rat Testicles; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with H3-3A | Histone H3.3 (p-S22) Antibody, Unconjugated (bsm-63099R) at 1:200 overnight at 4\u00b0C. Followed by conjugated Goat Anti-Rabbit IgG antibody (Red, bs-0295G-BF594), DAPI (blue, C02-04002) was used to stain the cell nuclei.
Paraformaldehyde-fixed, paraffin embedded Rat Testicles; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with H3-3A | Histone H3.3 (p-S22) Antibody, Unconjugated (bsm-63099R) at 1:200 overnight at 4\u00b0C. Followed by conjugated Goat Anti-Rabbit IgG antibody (Red, bs-0295G-BF594), DAPI (blue, C02-04002) was used to stain the cell nuclei.
Paraformaldehyde-fixed, paraffin embedded Mouse Testicles; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with H3-3A | Histone H3.3 (p-S22) Antibody, Unconjugated (bsm-63099R) at 1:200 overnight at 4\u00b0C. Followed by conjugated Goat Anti-Rabbit IgG antibody (Red, bs-0295G-BF594), DAPI (blue, C02-04002) was used to stain the cell nuclei.
Paraformaldehyde-fixed, paraffin embedded Mouse Testicles; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with H3-3A | Histone H3.3 (p-S22) Antibody, Unconjugated (bsm-63099R) at 1:200 overnight at 4\u00b0C. Followed by conjugated Goat Anti-Rabbit IgG antibody (Red, bs-0295G-BF594), DAPI (blue, C02-04002) was used to stain the cell nuclei.
Bioss unconjugated antibody vial

Applications

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

Reactivity

  • Human
  • Mouse
  • Rat
Overview
Catalog # bsm-63099r-100uL
Product Name Histone H3.3(S22) Monoclonal Antibody
Applications
WB IHC-P IHC-F IF FLOW-CYT ICC/IF
Reactivity Human, Mouse, Rat
Specifications
Conjugation Unconjugated
Host Rabbit
Source A synthesized peptide derived from human Histone H3.3 around the phosphorylation site of S22
Clonality Recombinant
Clone # 5D12
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 P84243
Subcellular location Cytoplasm, Nucleus
Synonyms H3-3A | Histone H3.3 (p-S22); p-Histone H3.3; phospho-Histone H3.3; H3.3S22P; p-H3.3; BRYLIB1; H3.3A; H3F3; H3F3A; BRYLIB2; H3.3B; H3F3B; H3-3b; H33_HUMAN; H3-3A; H33_RAT;
Application Dilution
WB =1:500-2000
IHC-P IHC-P=1:100-200
IHC-F IHC-F=1:100-200
IF IF=1:50-200
Flow-Cyt Flow-Cyt=1:50-100
ICC/IF ICC/IF=1:50-200

Frequently asked questions

Which applications is Histone H3.3(S22) Monoclonal Antibody (bsm-63099R) validated for, and at what dilution?

bsm-63099R is validated for WB (WB=1:500-2000); IHC-P (IHC-P=1:100-200); IHC-F (IHC-F=1:100-200); IF (IF=1:50-200); Flow-Cyt (Flow-Cyt=1:50-100); ICC/IF (ICC/IF=1:50-200). These are recommended starting dilutions; optimal dilutions should be determined by the end user.

Which species does Histone H3.3(S22) Monoclonal Antibody (bsm-63099R) react with?

Tested reactivity: Human, Mouse, Rat.

How should bsm-63099R 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 Histone H3.3(S22) Monoclonal Antibody (bsm-63099R)?

A synthesized peptide derived from human Histone H3.3 around the phosphorylation site of S22

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