bs-0349R [Primary Antibody]
Rabbit  Anti-Histone H3 Rabbit pAb, Nuclear Loading Control  Polyclonal Antibody
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Host: Rabbit

Target Protein: Histone H3 Rabbit pAb, Nuclear Loading Control

IR: Immunogen Range:71-136/136

Clonality: Polyclonal

Isotype: IgG

Entrez Gene: 8968

Swiss Prot: Q71DI3

Source: KLH conjugated synthetic peptide derived from human Histone H3:71-136/136 

Purification: affinity purified by Protein A

Storage: 0.01M TBS (pH7.4) with 1% BSA, 0.02% Proclin300 and 50% Glycerol. Shipped at 4℃. Store at -20℃ for one year. Avoid repeated freeze/thaw cycles.

Background: Modulation of the chromatin structure plays an important role in the regulation of transcription in eukaryotes. The nucleosome, made up of four core histone proteins (H2A, H2B, H3 and H4), is the primary building block of chromatin. The N-terminal tail of core histones undergoes different posttranslational modifications including acetylation, phosphorylation and methylation. These modifications occur in response to cell signal stimuli and have a direct effect on gene expression. In most species, the histone H2B is primarily acetylated at lysines 5, 12, 15 and 20. Histone H3 is primarily acetylated at lysines 9, 14, 18 and 23. Acetylation at lysine 9 appears to have a dominant role in histone deposition and chromatin assembly in some organisms. Phosphorylation at Ser10 of histone H3 is tightly correlated with chromosome condensation during both mitosis and meiosis.

Size: 500ul

Concentration: 1mg/ml

Applications: WB=1:5000-50000,IHC-P=1:500-2000,IHC-F=1:500-2000,IF=1:500-2000,Flow-Cyt=1μg/Test

Cross Reactive Species: Human,Mouse,Rat (predicted: Rabbit,Pig,Cow,Fruit Fly)

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

VALIDATION IMAGES
25 ug total protein per lane of various lysates (see on figure) probed with Histone H3 polyclonal antibody, unconjugated (bs-0349R) at 1:1000 dilution and 4°C overnight incubation. Followed by conjugated secondary antibody incubation at r.t. for 60 min.
Paraformaldehyde-fixed, paraffin embedded (rat brain); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (Histone H3HIST3H3(Nuclear Loading Control)) Polyclonal Antibody, Unconjugated (bs-0349R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (mouse liver); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (Histone H3 (Nuclear Loading Control) ) Polyclonal Antibody, Unconjugated (bs-0349R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (rat liver); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (Histone H3 (Nuclear Loading Control) ) Polyclonal Antibody, Unconjugated (bs-0349R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (mouse testis); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (Histone H3 (Nuclear Loading Control) ) Polyclonal Antibody, Unconjugated (bs-0349R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded Human Testicles; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with Histone H3 Polyclonal Antibody, Unconjugated (bs-0349R) at 1:1000 overnight at 4°C, followed by conjugation to the bs-0295G-HRP 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 Polyclonal Antibody, Unconjugated (bs-0349R) at 1:1000 overnight at 4°C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded (rat kidney); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (Histone H3HIST3H3(Nuclear Loading Control)) Polyclonal Antibody, Unconjugated (bs-0349R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (human laryngeal carcinoma); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (Histone H3HIST3H3(Nuclear Loading Control)) Polyclonal Antibody, Unconjugated (bs-0349R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (mouse kidney); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (Histone H3(Nuclear Loading Control)) Polyclonal Antibody, Unconjugated (bs-0349R ) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (mouse brain); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (Histone H3(Nuclear Loading Control)) Polyclonal Antibody, Unconjugated (bs-0349R ) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (mouse pancreas); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (Histone H3(Nuclear Loading Control)) Polyclonal Antibody, Unconjugated (bs-0349R ) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (rat pancreas); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (Histone H3(Nuclear Loading Control)) Polyclonal Antibody, Unconjugated (bs-0349R ) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (human colon); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (Histone H3(Nuclear Loading Control)) Polyclonal Antibody, Unconjugated (bs-0349R ) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (rat colon); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (Histone H3 (Nuclear Loading Control)) Polyclonal Antibody, Unconjugated (bs-0349R) at 1:300 overnight at 4°C, followed by a conjugated Goat Anti-Rabbit IgG antibody (YF488) for 90 minutes, and DAPI for nuclei staining.
Paraformaldehyde-fixed, paraffin embedded (human brain); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (Histone H3 (Nuclear Loading Control)) Polyclonal Antibody, Unconjugated (bs-0349R) at 1:500 overnight at 4°C, followed by a conjugated Goat Anti-Rabbit IgG antibody (YF488) for 90 minutes, and DAPI for nuclei staining.
Paraformaldehyde-fixed, paraffin embedded (rat testis); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (Histone H3 (Nuclear Loading Control)) Polyclonal Antibody, Unconjugated (bs-0349R) at 1:500 overnight at 4°C, followed by a conjugated Goat Anti-Rabbit IgG antibody (YF488) for 90 minutes, and DAPI for nuclei staining.
Paraformaldehyde-fixed, paraffin embedded (mouse testis); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (Histone H3 (Nuclear Loading Control)) Polyclonal Antibody, Unconjugated (bs-0349R) at 1:300 overnight at 4°C, followed by a conjugated Goat Anti-Rabbit IgG antibody (YF488) for 90 minutes, and DAPI for nuclei staining.
Paraformaldehyde-fixed, paraffin embedded (rat brain); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (Histone H3 (Nuclear Loading Control)) Polyclonal Antibody, Unconjugated (bs-0349R) at 1:300 overnight at 4°C, followed by a conjugated Goat Anti-Rabbit IgG antibody (YF488) for 90 minutes, and DAPI for nuclei staining.
Paraformaldehyde-fixed, paraffin embedded (mouse brain); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (Histone H3 (Nuclear Loading Control)) Polyclonal Antibody, Unconjugated (bs-0349R) at 1:300 overnight at 4°C, followed by a conjugated Goat Anti-Rabbit IgG antibody (YF488) for 90 minutes, and DAPI for nuclei staining.
Paraformaldehyde-fixed, paraffin embedded (human liver); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (Histone H3 (Nuclear Loading Control)) Polyclonal Antibody, Unconjugated (bs-0349R) at 1:300 overnight at 4°C, followed by a conjugated Goat Anti-Rabbit IgG antibody (YF488) for 90 minutes, and DAPI for nuclei staining.
Paraformaldehyde-fixed, paraffin embedded (mouse kidney); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (Histone H3 (Nuclear Loading Control)) Polyclonal Antibody, Unconjugated (bs-0349R) at 1:300 overnight at 4°C, followed by a conjugated Goat Anti-Rabbit IgG antibody (YF488) for 90 minutes, and DAPI for nuclei staining.
Paraformaldehyde-fixed, paraffin embedded (Human kidney); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (Histone H3 (Nuclear Loading Control)) Polyclonal Antibody, Unconjugated (bs-0349R) at 1:300 overnight at 4°C, followed by a conjugated Goat Anti-Rabbit IgG antibody (YF488) for 90 minutes, and DAPI for nuclei staining.
Paraformaldehyde-fixed, paraffin embedded (rat kidney); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (Histone H3 (Nuclear Loading Control)) Polyclonal Antibody, Unconjugated (bs-0349R) at 1:300 overnight at 4°C, followed by a conjugated Goat Anti-Rabbit IgG antibody (YF488) for 90 minutes, and DAPI for nuclei staining.
Paraformaldehyde-fixed, paraffin embedded (rat liver); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (Histone H3 (Nuclear Loading Control)) Polyclonal Antibody, Unconjugated (bs-0349R) at 1:300 overnight at 4°C, followed by a conjugated Goat Anti-Rabbit IgG antibody (YF488) for 90 minutes, and DAPI for nuclei staining.
Paraformaldehyde-fixed, paraffin embedded (mouse liver); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (Histone H3 (Nuclear Loading Control)) Polyclonal Antibody, Unconjugated (bs-0349R) at 1:300 overnight at 4°C, followed by a conjugated Goat Anti-Rabbit IgG antibody (YF488) for 90 minutes, and DAPI for nuclei staining.
Blank control: K562. Primary Antibody (green line): Rabbit Anti-Histone H3/HIST3H3 antibody (bs-0349R) Dilution: 1μg /10^6 cells; Isotype Control Antibody (orange line): Rabbit IgG . Secondary Antibody : Goat anti-rabbit IgG-PE Dilution: 1μg /test. Protocol The cells were fixed with 4% PFA (10min at room temperature)and then permeabilized with 90% ice-cold methanol for 20 min at-20℃. The cells were then incubated in 5%BSA to block non-specific protein-protein interactions for 30 min at at room temperature .Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20,000 events was performed.
Blank control: Mouse spleen cells (blue). Primary Antibody:Rabbit Anti-Histone H3/HIST3H3 antibody(bs-0349R), Dilution: 1μg in 100 μL 1X PBS containing 0.5% BSA; Isotype Control Antibody: Rabbit IgG(orange) ,used under the same conditions ); Secondary Antibody: Goat anti-rabbit IgG-PE(white blue), Dilution: 1:200 in 1 X PBS containing 0.5% BSA. Protocol The cells were fixed with 2% paraformaldehyde (10 min). Primary antibody (bs-0349R, 1μg /1x10^6 cells) were incubated for 30 min on the ice, followed by 1 X PBS containing 0.5% BSA + 1 0% goat serum (15 min) to block non-specific protein-protein interactions. Then the Goat Anti-rabbit IgG/PE antibody was added into the blocking buffer mentioned above to react with the primary antibody at 1/200 dilution for 30 min on ice. Acquisition of 20,000 events was performed.
 
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