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Histone H3 Ready-To-Use IHC Kit (IHC0102)  
訂購熱線:400-901-9800
訂購郵箱:sales@bioss.com.cn
訂購QQ:  400-901-9800
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說明書: 下載
50T/3200.00元
大包裝/詢價

產品編號 IHC0102
英文名稱 Histone H3 Ready-To-Use IHC Kit
中文名稱 組蛋白H3(核內參)即用型免疫組化試劑盒
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  
種    屬 Human, Mouse, Rat
應用范圍 Immunohistochemistry
適用樣品基質 PFPE tissue
描述 IHC0102 is a ready-to-use IHC kit for staining of Histone H3. The kit provides all reagents, from antigen retrieval to cover slip mounting, that require little to no diluting or handling prior to use. Simply apply the reagents to your sample slide according to the protocol and you're steps away from obtaining high-quality IHC data.
保存條件 Please store components at the temperatures indicated on the individual tube labels. The kit is stable for 6 months from the date of receipt.
注意事項 This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications.
產品介紹


產品組分及規(guī)格

編號

組分

規(guī)格

濃度

儲存

1

PBS緩沖液干粉

2 L×2

20x

室溫

2

抗原修復緩沖液

20 ml

100x

2-8

3

內源性過氧化物酶阻斷劑

3 ml

RTU

2-8

4

封閉工作液

3 ml

RTU

2-8

5

一抗Histone H3 Rabbit pAb

6 ml

RTU

2-8

6

二抗HRP-Goat anti-Rabbit IgG pAb

6 ml

RTU

2-8

7

DAB kit20×)顯色液

0.3 ml

RTU

-20℃

8

DAB kit20×)稀釋液

0.3 ml

RTU

-20

9

復染試劑

5 ml

RTU

室溫

10

封片劑

5 ml

RTU

室溫

11

對照大鼠肝,小鼠肝,人腎

3

RTU

室溫

12

說明書

1


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圍畫圈,加入2-4內源性過氧化物酶阻斷劑(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2-4Histone H3 兔多抗工作液(試劑5,置于濕盒中,4℃孵育過夜或37℃孵育1-2 h。

6. 復溫

4℃孵育過夜后,室溫下復溫15 min (若在室溫下孵育一抗,則直接進入下一步清洗);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2-4HRP標記羊抗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。

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,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,避免產生氣泡。


注意事項

1. 建議檢測時進行陰性及陽性對照,以提高實驗的可靠性。

2. 本品中的配套試劑,請不要用其他生產商產品替換使用。

3. DAB為致癌物質,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78)的工作液需每次實驗時現用現配。

*5. 發(fā)表論文時引用本產品的寫作建議 "IHC0102, Bioss Antibodies"。引用示例: Tissue sections using Histone H3 IHC Kit (IHC0102, Bioss Antibodies) were stained for Histone H3 according to the manufacturer's instructions.”

產品圖片
Immunohistochemical analysis of paraffin embedded mouse liver tissue slide using IHC0102 (Histone H3 IHC Kit).
Immunohistochemical analysis of paraffin embedded rat liver tissue slide using IHC0102 (Histone H3 IHC Kit).
Immunohistochemical analysis of paraffin embedded human kidney tissue slide using IHC0102 (Histone H3 IHC Kit).
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