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免疫组化求助

免疫组化求助

我有一个一抗,主要用于人的组织,但与多种种属有交叉反应。
已经做过实验一抗1.5:500稀释,人皮肤的石蜡切片有阳性细胞着色,而大鼠皮肤的石蜡切片却看不到阳性细胞
请问各位老师,我该如何调整实验条件?
(要增加一抗的浓度吗?
要增加一抗孵育的时间吗?
还有其他要改进的吗?)
一抗的说明书(……Species Cross-reactivity: Human.Predicted to crossreact
with many species based on extensive sequence homology.) 

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不同种属组织中的最佳浓度不尽相同有可能出现有在人体组织中用较小的浓度可以显示但其他种属要用高浓度的这种情况,而且用一个大鼠组织来测试抗体最佳浓度不是太恰当。建议多增加几个测试样本。你如果可能的话把抗体具体名称、来源说清楚,我们好帮你。

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谢谢老师!
一抗为
Anti-Tyrosinase,clone T311
(mouse monoclonal IgG2α)
Formulation:200ug og protein G purified mouse IgG2α in 232ul of  0.1MTris-glycine ,PH 7.4 ,0.15M Nacl,0.05% sodium azide before the addition of glycerol to 30%,Liquid at -20摄氏度。
我主要是要测大鼠皮肤的Tyrosinase,但没有阳性细胞显示。

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Anti-Tyrosinase, clone T311  
Host: Mouse

Antibody Type: Monoclonal Antibody

Isotype: IgG2a

Method of Purification: Protein G chromatography

Immunogen:
Purified recombinant Tyrosinase corresponding to residues 5-456 of human Tyrosinase

Specificity: Tyrosinase

GenBank Accession # (click for details from Entrez Gene):
NM_000372

Molecular Weight: 70-80kDa

Quality Assurance:
routinely evaluated by immunoblot on RIPA lysates from SK-MEL-19 cells

Presentation:
0.1M Tris-glycine, pH 7.4, 0.15M NaCl, 0.05% sodium azide before the addition of glycerol to 30%

Storage/Handling: 2 years at -20°C

Tested Applications: Immunoblotting (Western), Immunohistochemistry (tissue)

Application Notes:
Not recommended for Immunoprecipitation

Species Reactivity:  Human. Reactivity with other species has not been confirmed. (请注意最后一行话)

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谢谢老师的指点!

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回复

王bobo同志你的免疫组化做出来了吗研究生的课题做好吗???

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