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組蛋白H3(核內參)抗體

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中文名稱 組蛋白H3(核內參)抗體
別    名 H3 histone family member E pseudogene; H3F3; HIST3H3; Histone H3 3 pseudogene; H31_TETTH; Histone H3; H3S; Histone H3-I/H3-II; Major histone H3; H3F; 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.  

 

產品類型 內參抗體 
研究領域 免疫學  細胞周期蛋白  
抗體來源 Rabbit
克隆類型 Polyclonal
交叉反應 Human, Mouse, Rat,  (predicted: Pig, Cow, Rabbit, Fruit Fly, )
產品應用 WB=1:500-2000 ELISA=1:500-1000 IHC-P=1:100-500 IHC-F=1:100-500 Flow-Cyt=1μg/Test ICC=1:100 IF=1:100-500 (石蠟切片需做抗原修復)
not yet tested in other applications.
optimal dilutions/concentrations should be determined by the end user.
分 子 量 15kDa
細胞定位 細胞核 
性    狀 Liquid
濃    度 1mg/ml
免 疫 原 KLH conjugated synthetic peptide derived from human Histone H3:71-136/136 
亞    型 IgG
純化方法 affinity purified by Protein A
儲 存 液 0.01M TBS(pH7.4) with 1% BSA, 0.03% Proclin300 and 50% Glycerol.
保存條件 Shipped at 4℃. Store at -20 °C for one year. Avoid repeated freeze/thaw cycles.
PubMed PubMed
產品介紹 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.

Function:
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.

Subunit:
The nucleosome is a histone octamer containing two molecules each of H2A, H2B, H3 and H4 assembled in one H3-H4 heterotetramer and two H2A-H2B heterodimers. The octamer wraps approximately 147 bp of DNA. Interacts with GCN5, whereby H3S10ph increases histone-protein interactions. Interacts with PDD1 and PDD3.

Subcellular Location:
Nucleus. Chromosome. Note=Localizes to both the large, transcriptionally active, somatic macronucleus (MAC) and the small, transcriptionally inert, germ line micronucleus (MIC).

Post-translational modifications:
Phosphorylated to form H3S10ph. H3S10ph promotes subsequent H3K14ac formation by GCN5. H3S10ph is only found in the mitotically dividing MIC, but not in the amitotically dividing MAC. H3S10ph is correlated with chromosome condensation during mitotic or meiotic micronuclear divisions.
Acetylation of histone H3 leads to transcriptional activation. H3K14ac formation by GCN5 is promoted by H3S10ph. H3K9acK14ac is the preferred acetylated form of newly synthesized H3. Acetylation occurs almost exclusively in the MAC.
Methylated to form H3K4me. H3K4me is only found in the transcriptionally active MAC. Methylated to form H3K9me in developing MACs during conjugation, when genome-wide DNA elimination occurs. At this stage, H3K9me specifically occurs on DNA sequences being eliminated (IES), probably targeted by small scan RNAs (scnRNAs) bound to IES, and is required for efficient IES elimination. H3K9me is required for the interaction with the chromodomains of PDD1 and PDD3.
The full-length protein H3S (slow migrating) is converted to H3F (fast migrating) by proteolytic removal of the first 6 residues. H3F is unique to MIC, and processing seems to occur regularly each generation at a specific point in the cell cycle.

Similarity:
Belongs to the histone H3 family.

SWISS:
P84243

Gene ID:
8290

Database links:

Entrez Gene: 326601 Cow

Entrez Gene: 8350 Human

Entrez Gene: 8351 Human

Entrez Gene: 8352 Human

Entrez Gene: 8353 Human

Entrez Gene: 8354 Human

Entrez Gene: 8355 Human

Entrez Gene: 8290 Human

 

Entrez Gene: 8350 Human

 

Entrez Gene: 8351 Human

 

Entrez Gene: 8352 Human

 

Entrez Gene: 8353 Human

 

Entrez Gene: 8354 Human

 

Entrez Gene: 8355 Human

 

Entrez Gene: 8356 Human

 

Entrez Gene: 8357 Human

 

Entrez Gene: 8358 Human

 

Entrez Gene: 8968 Human

 

Entrez Gene: 260423 Mouse

 

Entrez Gene: 319148 Mouse

 

Entrez Gene: 319149 Mouse

 

Entrez Gene: 319150 Mouse

 

Entrez Gene: 319151 Mouse

 

Entrez Gene: 319152 Mouse

 

Entrez Gene: 319153 Mouse

 

Entrez Gene: 360198 Mouse

 

Entrez Gene: 97908 Mouse

 

Entrez Gene: 100364501 Rat

 

Entrez Gene: 100365669 Rat

 

Entrez Gene: 291159 Rat

 

Entrez Gene: 314977 Rat

 

Entrez Gene: 364716 Rat

 

Entrez Gene: 679950 Rat

 

Entrez Gene: 679994 Rat

 

Entrez Gene: 680511 Rat

 

Entrez Gene: 680599 Rat

 

Entrez Gene: 682330 Rat

 

Entrez Gene: 691496 Rat

 

Omim: 601128 Human

 

Omim: 602810 Human

 

Omim: 602811 Human

 

Omim: 602812 Human

 

Omim: 602813 Human

 

Omim: 602814 Human

 

Omim: 602815 Human

 

Omim: 602816 Human

 

Omim: 602817 Human

 

Omim: 602818 Human

 

Omim: 602819 Human

 

SwissProt: P68431 Human

 

SwissProt: P84243 Human

 

SwissProt: Q16695 Human

 

SwissProt: Q6NXT2 Human

 

SwissProt: Q71DI3 Human

 

SwissProt: P68433 Mouse

 

SwissProt: P84228 Mouse

 

SwissProt: Q6LED0 Rat

 

Unigene: 132854 Human

 

Unigene: 247813 Human

 

Unigene: 247814 Human

 

Unigene: 248176 Human

 

Unigene: 443021 Human

 

Unigene: 484990 Human

 

Unigene: 532144 Human

 

Unigene: 533292 Human

 

Unigene: 546315 Human

 

Unigene: 586261 Human

 

Unigene: 591778 Human

 

Unigene: 221301 Mouse

 

Unigene: 261657 Mouse

 

Unigene: 377874 Mouse

 

Unigene: 390558 Mouse

 

Unigene: 397328 Mouse

 

Unigene: 138090 Rat

 



Important Note:
This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications.

組蛋白的基因非常保守,在親緣關系較遠的種屬中,四種組蛋白(H2A、H2A、H3、H4)氨基酸序列都非常相似,如海膽組織H3的氨基酸序列與來自小牛胸腺的H3的氨基酸序列間只有一個氨基酸的差異,小牛胸腺的H3的氨基酸序列與豌豆的H3也很相似。組蛋白是細胞核內的一種堿性核蛋白,抗組蛋白抗體即是以組蛋白為靶抗原的一種自身,是抗核抗體的一種。分子量:16-18KDa。主要與藥物性紅斑狼瘡、系統(tǒng)性紅斑狼瘡、類風濕關節(jié)炎有關。
產品圖片 Sample: K562 Cell Lysate at 40 ug
Primary: Anti-Histone H3/HIST3H3(bs-0349R) at 1/300 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/10000 dilution
Predicted band size: 15kD
Observed band size: 17kD
Sample:
Lane 1: NIH/3T3(Mouse) Cell Lysate at 30 ug
Lane 2: MCF-7 (Human) Cell Lysate at 30 ug
Lane 3: SiHa (Human) Cell Lysate at 30 ug
Lane 4: U-2OS (Human) Cell Lysate at 30 ug
Lane 5: MOLT-4 (Human) Cell Lysate at 30 ug
Primary: Anti-Histone H3’HIST3H3 (bs-0349R) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 17 kD
Observed band size: 15 kD
Sample:
293T(Human) Cell Lysate at 30 ug
293T(invent)(Human) Cell Lysate at 30 ug
Primary: Anti-HIST3H3  (bs-0349R) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 15 kD
Observed band size: 15 kD
Sample:
K562(Human) Cell Lysate at 30 ug
Primary: Anti-Histone H3/HIST3H3 (bs-0349R) at 1/300 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 15 kD
Observed band size: 17 kD
Paraformaldehyde-fixed, paraffin embedded (human cervical 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 (HIST3H3) Polyclonal Antibody, Unconjugated (bs-0349R) at 1:400 overnight at 4°C, followed by a conjugated secondary (sp-0023) for 20 minutes and DAB staining.Paraformaldehyde-fixed, paraffin embedded (human colon cancer); 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 (HIST3H3) Polyclonal Antibody, Unconjugated (bs-0349R) at 1:400 overnight at 4°C, followed by a conjugated secondary (sp-0023) for 20 minutes and DAB staining.Paraformaldehyde-fixed, paraffin embedded (Human 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 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 (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 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 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 (rat 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 (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 (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 (Mouse small intestine); 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 (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 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; 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 stomach); 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) Polyclonal Antibody, Unconjugated (bs-0349R) at 1:400 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.Tissue/cell: rat brain tissue;4% Paraformaldehyde-fixed and paraffin-embedded;
Antigen retrieval: citrate buffer ( 0.01M, pH 6.0 ), Boiling bathing for 15min; Blocking buffer (normal goat serum,C-0005) at 37℃ for 20 min;
Incubation: Anti-Histone H3 Polyclonal Antibody, FITC conjugated (bs-0349R-FITC) used at 1:100 dilution for 40 minutes at 37癈.
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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