| 產(chǎn)品編號 | bsm-33042R | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| 英文名稱(chēng) | Histone H3 Recombinant Rabbit mAb | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| 中文名稱(chēng) | 組蛋白H3（核內參）重組兔單抗 | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| 別 名 | 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; | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| 產(chǎn)品類(lèi)型 | 內參抗體 重組兔單抗 | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| 抗體來(lái)源 | Rabbit | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| 克隆類(lèi)型 | Recombinant | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| 克 隆 號 | 3G1 | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| 交叉反應 | Human,Mouse,Rat (predicted: Hamster,Bee) | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| 產(chǎn)品應用 | WB=1:2000-20000,IHC-P=1:200-1000,IHC-F=1:200-1000,IF=1:200-1000,Flow-Cyt=1μg/Test,ICC/IF=1:50-200
not yet tested in other applications. optimal dilutions/concentrations should be determined by the end user. |
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| 理論分子量 | 15 kDa | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| 檢測分子量 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| 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 | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| 亞 型 | IgG | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| 純化方法 | affinity purified by Protein A | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| 緩 沖 液 | 0.01M TBS (pH7.4) with 1% BSA, 0.02% Proclin300 and 50% Glycerol. | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| 保存條件 | Shipped at 4℃. Store at -20℃ for one year. Avoid repeated freeze/thaw cycles. | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| 注意事項 | This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications. | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| PubMed | PubMed | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| 產(chǎn)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). Tissue Specificity: Expressed in testicular cells.Expressed during S phase, then expression strongly decreases as cell division slows down during the process of differentiation. 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. DISEASE: Expressed during S phase, then expression strongly decreases as cell division slows down during the process of differentiation. Similarity: Belongs to the histone H3 family. SWISS: Q71DI3 Gene ID: 8968 Database links: 產(chǎn)品組分及規格
背景 Histone H3 is one of the DNA-binding proteins found in the chromatin of all eukaryotic cells. H3 along with four core histone proteins binds to DNA forming the structure of the nucleosome. Histones play a central role in transcription regulation, DNA repair, DNA replication and chromosomal stability. Post translationally, histones are modified in a variety of ways to either directly change the chromatin structure or allow for the binding of specific transcription factors. The N-terminal tail of histone H3 protrudes from the globular nucleosome core and can undergo several different types of post-translational modification that influence cellular processes. These modifications include the covalent attachment of methyl or acetyl groups to lysine and arginine amino acids and the phosphorylation of serine or threonine.
石蠟包埋組織的免疫組織化學(xué)方案 1. 脫蠟水化 石蠟切片置于新鮮二甲苯中浸泡脫蠟3次，每次15 min；依次置于不同濃度（100%、95%、90%、80%、70%）乙醇浸泡各5 min，再置于蒸餾水洗滌5 min，重復3次。 2. 抗原修復 沸水浴修復：將100×抗原修復緩沖液（試劑2）用蒸餾水稀釋成1×抗原修復緩沖液，放入修復盒中并提前加熱至95-100℃（注意蓋好以防液體蒸發(fā)），然后將切片放入修復盒中，在沸水浴環(huán)境中保持外沸狀態(tài)15 min，室溫自然冷卻；用PBS緩沖液（試劑1，將干粉溶解在2L蒸餾水中）清洗5 min，重復3次。 3. 阻斷內源性過(guò)氧化物酶 用吸水紙吸去玻片上多余的液體，用免疫組化筆在組織周?chē)?huà)圈，加入2-4滴內源性過(guò)氧化物酶阻斷劑（試劑3），室溫下置于濕盒中孵育15 min，用PBS洗滌5 min，重復3次。 4. 血清封閉 用吸水紙吸去玻片上多余的液體，加入2-4滴封閉工作液（試劑4），置于濕盒內37℃封閉20 min，以減少非特異性染色。 5. 一抗孵育 用吸水紙吸去玻片上多余的液體，加入2-4滴大鼠Histone H3 鼠單抗工作液（試劑5），置于濕盒中，4℃孵育過(guò)夜或37℃孵育1-2 h。 6. 復溫 4℃孵育過(guò)夜后，室溫下復溫15 min (若在室溫下孵育一抗，則直接進(jìn)入下一步清洗)；用PBS洗滌5 min，重復3次。 7. 二抗孵育 用吸水紙吸玻片上多余的液體，加入2-4滴HRP標記羊抗鼠IgG工作液（試劑6），置于濕盒中，37℃孵育1-2 h；用PBS洗滌5 min，重復3次。 8. 顯色 用吸水紙吸去玻片上多余的液體，在每張切片上滴加約50 μL新配制的DAB工作液（試劑7:試劑8:PBS=1:1:18），作用3-5 min。顯微鏡下觀(guān)察結果，達到合適的顯色強度后，用蒸餾水沖洗切片以終止反應，用蒸餾水沖洗5 min，重復3次。 9. 復染 滴加適量復染試劑（試劑9）復染3-5 min，蒸餾水沖洗5 min，滴加鹽酸酒精分化約30 s，蒸餾水洗滌5 min，重復2次。 10. 脫水封片 將玻片依次置于不同濃度（70%、80%、90%、95%、100%）乙醇，各5 min；然后置于新鮮二甲苯中浸泡脫蠟3次，每次15 min。用吸水紙吸去多余的二甲苯，滴加適量封片劑（試劑10）在組織上，將蓋玻片蓋在組織上，避免產(chǎn)生氣泡。
注意事項 1. 建議檢測時(shí)進(jìn)行陰性及陽(yáng)性對照，以提高實(shí)驗的可靠性。 2. 本品中的配套試劑，請不要用其他生產(chǎn)商產(chǎn)品替換使用。 3. DAB為致癌物質(zhì)，請采取必要的防范措施。 4. PBS洗滌液（試劑1）配制后在4℃可保存一周；抗原修復液（試劑2）及顯色劑（試劑7和8）的工作液需每次實(shí)驗時(shí)現用現配。 *5. 發(fā)表論文時(shí)引用本產(chǎn)品的寫(xiě)作建議 "IHC0111R, Bioss Antibodies"。引用示例: “Rat tissue sections using Rat Histone H3 IHC Kit (IHC0111R, Bioss Antibodies) were stained for Histone H3 according to the manufacturer's instructions.” |
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| 產(chǎn)品圖片 |
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.
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.
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 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 Liver 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 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 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 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 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 Human Cerebrum; 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 Cerebrum; 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 Cerebrum; 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 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 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 Human Testicles; 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 Testicles; 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 Testicles; 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 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 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.
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.
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.
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.
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