bsm-33042R [Primary Antibody]
Histone H3 Monoclonal Antibody
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Host: Rabbit

Target Protein: Histone H3

Clonality: Monoclonal

Isotype: IgG

Entrez Gene: 8968

Swiss Prot: Q71DI3

Source: KLH conjugated synthetic peptide derived from human Histone H3

Purification: Purified by Protein A.

Storage Buffer: 0.01M TBS(pH7.4) with 1% BSA, 0.02% Proclin300 and 50% Glycerol.

Storage: Shipped at 4C. Store at -20C for one year. Avoid repeated freeze/thaw cycles.

Background:

Core component of nucleosome. Nucleosomes wrap and compact DNA into chromatin, limiting DNA accessibility to the cellular machineries which require DNA as a template. Histones thereby play a central role in transcription regulation, DNA repair, DNA replication and chromosomal stability. DNA accessibility is regulated via a complex set of post-translational modifications of histones, also called histone code, and nucleosome remodeling. H3 is deposited into chromatin exclusively through a DNA replication-coupled pathway that can be associated with either DNA duplication or DNA repair synthesis during meiotic homologous recombination.

Size: 100µL

Concentration: 1ug/ul

Applications: WB(WB(1:5000-50000))
FCM(FCM(1μg/Test))
IHC-P(IHC-P(1:500-2000))
IHC-F(IHC-F(1:500-2000))
IF(IHC-P)(IF(IHC-P)(1:500-2000))
IF(ICC)(IF(ICC)(1:100-500))
ChIP(ChIP(6μg/IP))

Predicted Molecular Weight: 15


Cross Reactive Species: Human
Mouse
Rat

Predicted Cross Reactive Species: Bee
Hamster

For research use only. Not intended for diagnostic or therapeutic use.

VALIDATION IMAGES

Paraformaldehyde-fixed, paraffin embedded Rat Liver; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with Histone H3 Monoclonal Antibody, Unconjugated(bsm-33042R) at 1:200 overnight at 4°C, followed by conjugation to the SP Kit(Rabbit, SP-0023) and DAB (C-0010) staining.


Paraformaldehyde-fixed, paraffin embedded Mouse Liver; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with Histone H3 Monoclonal Antibody, Unconjugated(bsm-33042R) at 1:200 overnight at 4°C, followed by conjugation to the SP Kit(Rabbit, SP-0023) and DAB (C-0010) staining.


Paraformaldehyde-fixed, paraffin embedded Human Liver; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with Histone H3 Monoclonal Antibody, Unconjugated(bsm-33042R) at 1:200 overnight at 4°C, followed by conjugation to the SP Kit(Rabbit, SP-0023) and DAB (C-0010) staining.


Paraformaldehyde-fixed, paraffin embedded Human Colon; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with Histone H3 Monoclonal Antibody, Unconjugated(bsm-33042R) at 1:200 overnight at 4°C, followed by conjugation to the SP Kit(Rabbit, SP-0023) and DAB (C-0010) staining.


Paraformaldehyde-fixed, paraffin embedded Rat Colon; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with Histone H3 Monoclonal Antibody, Unconjugated(bsm-33042R) at 1:200 overnight at 4°C, followed by conjugation to the SP Kit(Rabbit, SP-0023) and DAB (C-0010) staining.


Paraformaldehyde-fixed, paraffin embedded Mouse Colon; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with Histone H3 Monoclonal Antibody, Unconjugated(bsm-33042R) at 1:200 overnight at 4°C, followed by conjugation to the SP Kit(Rabbit, SP-0023) and DAB (C-0010) staining.


Paraformaldehyde-fixed, paraffin embedded Mouse Kidney; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with Histone H3 Monoclonal Antibody, Unconjugated(bsm-33042R) at 1:200 overnight at 4°C, followed by conjugation to the SP Kit(Rabbit, SP-0023) and DAB (C-0010) staining.


Paraformaldehyde-fixed, paraffin embedded Rat Kidney; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with Histone H3 Monoclonal Antibody, Unconjugated(bsm-33042R) at 1:200 overnight at 4°C, followed by conjugation to the SP Kit(Rabbit, SP-0023) and DAB (C-0010) staining.


Paraformaldehyde-fixed, paraffin embedded Rat Thymus; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with Histone H3 Monoclonal Antibody, Unconjugated(bsm-33042R) at 1:200 overnight at 4°C, followed by conjugation to the SP Kit(Rabbit, SP-0023) and DAB (C-0010) staining.


Paraformaldehyde-fixed, paraffin embedded Mouse Thymus; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with Histone H3 Monoclonal Antibody, Unconjugated(bsm-33042R) at 1:200 overnight at 4°C, followed by conjugation to the SP Kit(Rabbit, SP-0023) and DAB (C-0010) staining.


Paraformaldehyde-fixed, paraffin embedded Human Lung Cancer; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with Histone H3 Monoclonal Antibody, Unconjugated(bsm-33042R) at 1:200 overnight at 4°C, followed by conjugation to the SP Kit(Rabbit, SP-0023) 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; Antibody incubation with Histone H3 Monoclonal Antibody, Unconjugated(bsm-33042R) at 1:200 overnight at 4°C, followed by conjugation to the SP Kit(Rabbit, SP-0023) and DAB (C-0010) staining.


Paraformaldehyde-fixed, paraffin embedded Human Pancreas; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with Histone H3 Monoclonal Antibody, Unconjugated(bsm-33042R) at 1:200 overnight at 4°C, followed by conjugation to the SP Kit(Rabbit, SP-0023) and DAB (C-0010) staining.


Paraformaldehyde-fixed, paraffin embedded Mouse Colon; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with Histone H3 Monoclonal Antibody, Unconjugated (bsm-33042R) at 1:200 overnight at 4°C. Followed by conjugated Goat Anti-Rabbit IgG antibody (green, bs-0295G-BF488), DAPI (blue, C02-04002) was used to stain the cell nuclei.


Paraformaldehyde-fixed, paraffin embedded Rat Colon; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with Histone H3 Monoclonal Antibody, Unconjugated (bsm-33042R) at 1:200 overnight at 4°C. Followed by conjugated Goat Anti-Rabbit IgG antibody (green, bs-0295G-BF488), DAPI (blue, C02-04002) was used to stain the cell nuclei.


Paraformaldehyde-fixed, paraffin embedded Human Kidney; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with Histone H3 Monoclonal Antibody, Unconjugated(bsm-33042R) at 1:200 overnight at 4°C, 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 monoclonal antibody, unconjugated (bsm-33042R) at 1:5000 dilution and 4°C overnight incubation. Followed by conjugated secondary antibody incubation at r.t. for 60 min.


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℃,the cells then were incubated in 5%BSA to block non-specific protein-protein interactions (30 min at r.t.).Primary Antibody (green):Rabbit Anti-Histone H3 antibody (bsm-33042R): 1 μg/10^6 cells; Secondary Antibody (white blue): Goat anti-Rabbit IgG-BF488 (bs-60295G-BF488): 1 μg/test. Blank control (black): PBS. Acquisition of 20,000 events was performed.


4% Paraformaldehyde-fixed Hela (H) cell; Triton X-100 at r.t. for 20 min; Antibody incubation with (Histone H3) monoclonal Antibody, unconjugated (bsm-33042R) 1:100, 90 min at 37°C; followed by conjugated Goat Anti-Rabbit IgG antibody (green, bs-60295G-BF488) at 37°C 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.


Chromatin immunoprecipitation was performed using cross-linked chromatin from 5×10⁶ HeLa cells per IP, with 6 μg of bsm-33042R antibody or 6 μg of normal rabbit IgG(negative control bs-0295P) , and 20 μL of Protein A/G MagBeads per IP. The immunoprecipitated DNA was quantified by real-time PCR using primers specific for the human GAPDH-CDS, RPL30 exon 4, LDHA-promoter and FOXO3A-promoter regions. The data are presented as enrichment of each sample relative to the total amount of input at each amplicon.