链酶亲和素（SAV）免疫磁珠

100ug (NP19865) Please inquiry

Antibody type:Flowcytometry

Label:Unconjugated

Modification:Unmodification

Isotype:Unknown

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Application:FCM

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Form:Liquid

Tested Applications:

Western blot (1:100 to 1:500)
Immunofluorescence (1:50 to 1:400)
Immunohistochemistry (1:200 to 1:500)
Flow cytometry analysis (1:200 to 1:500)
Enzyme-linked Immunosorbent Assay (1:100-1:5000)

Note: Users are strongly advised to determine the optimal dilution of antibody to use for their specific applications.

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Storage:2-8℃保存，请勿冻存、干燥、加热或离心操作

Buffer condition:PBS,BSA,EDTA,0.09% NaN3,pH7.4

Background:1. 使用淋巴细胞分离液从抗凝血中提取外周血单个核细胞（PBMC），或者从淋巴组织中制备单细胞悬液。PBMC用PBS, 5%BSA*, 0..09%NaN3, pH7.4洗3次， 40-70µm尼龙网过滤去除细胞团块和碎片后调整细胞数为2-3uf0b4107/ml。2. 1uf0b4107 PBMC中加入适量生物素化抗体，一般1uf0b4107 PBMC中加入0.5-10µg 抗体, 室温反应30min。3. 1000rpm离心5min，弃上清中未结合的生物素化抗体。加入1ml磁珠分选缓冲液重悬细胞，1000rpm离心5min，弃上清，洗2次（2uf0b41ml）。4. 1ml磁珠分选缓冲液重悬细胞，将细胞与50µl免疫磁珠混合, 室温反应30min，间隔10min轻轻晃动反应管，避免磁珠沉淀，使磁珠与细胞充分反应，尽量防止出现气泡。阴性分选采用步骤5-7，阳性分离采用步骤8-9。阴性分选 (Negative Selection)5. 磁分器分离5-8min，收集未被免疫磁珠结合的细胞悬液至另一干净管中。6. 加入1ml磁珠分选缓冲液重悬磁珠，用玻璃滴管或移液枪枪头反复吹打磁珠。7. 重复步骤5，合并两次收集的细胞悬液，可将细胞悬液再置磁分离器分离5-8min，可明显增加靶细胞纯度。阳性分选 (Positive Selection)8. 磁分离器分离5-8min，去除未与磁珠结合的细胞。9. 结合了靶细胞的免疫磁珠用1ml磁珠分选缓冲液洗5次（5uf0b41ml）后，可直接进行流式检测，或者37oC培养48hrs可使细胞与磁珠分离。

Molar Function:链亲和素(streptavidin)是一种由四个相同亚基组成的蛋白质，每个亚基均有一个与生物素有高亲和力(Kd 10-15 molar) 的结合位点。包被了链亲和素的磁珠，借链亲和素-生物素高亲和力结合特性，可与生物素化蛋白，多肽，及非蛋白质（如各种DNA、RNA分子）等分子结合，从混合体系中快速分离生物素化成分，进行如亲和纯化，细胞分离，DNA探针分析和mRNA分离等多方面应用。应用streptavidin磁珠可配合自己需要选择生物素标记的各种单抗去分离细胞，应用范围广泛，使用灵活。其原理是在实验系统中加入生物素标记的单抗，磁珠通过streptavidin与生物素标记的单抗结合后，单抗与细胞表面相应抗原特异性结合而使细胞被磁珠间接捕获，从而达到纯化分离细胞的目的。Streptavidin 磁珠可通过直接法或间接法进行细胞分选。直接法是先将抗特异细胞表面抗原的生物素化抗体与磁珠表面的streptavidin结合，待它与混合体系中的细胞反应后，利用磁力的作用，使与磁珠结合的细胞与其它细胞分开，达到纯化、分离的目的。间接法是将生物素化特异抗抗体先与混合体系中靶细胞表面特异抗原结合，再与streptavidin磁珠混合以达到细胞分选的目的。

Western Blot:

Stability: The thermal stability is described by the loss rate. The loss rate was determined by accelerated thermal degradation test, that is, incubate the protein at 37°C for 48h, and no obvious degradation and precipitation were observed. The loss rate is less than 5% within the expiration date under appropriate storage condition.

Storage: Store at 4°C for frequent use. Stored at -20°C in a manual defrost freezer for one year without detectable loss of activity. Avoid repeated freeze-thaw cycles.

Notes: For In vitro laboratory use only. Not for any clinical, therapeutic, or diagnostic use in humans or animals. Not for animal or human consumption.