WB, IHC
H, R, M, Mk, Dg, Ch, Hm, Rb, Pg, Sh
55KD
IgG1
PBS, pH 7.4, containing 0.02% Kathon as Preservative and 50% Glycerol. Store at -20°C.
Do not aliquot the antibody.
WB:1:5000
IHC:1:100-500
Antibodycandetectsendogenous β-tubulin protein
Microtubules are constituent parts of the mitotic apparatus, cilia, flagella, and elements of the cytoskeleton. They consist
principally of 2 soluble proteins, alpha- and beta-tubulin, each of about 55,000 Da. Antibodies against beta Tubulin are
useful as loading controls for Western Blotting. However it should be noted that levels of β-Tubulin may not be stable in
certain cells. For example, expression of β-Tubulin in adipose tissue is very low and therefore β-Tubulin should not be used
as loading control for these tissues.
更新中...
答:可能的原因有:
1、目的蛋白有多个修饰位点,本身可以呈现多条带,建议查阅文献或进行生物信息学分析,获得蛋白序列的修饰位点信息,通过去修饰确定蛋白实际大小;
2、样本处理过程中目的蛋白发生降解,建议加入蛋白酶抑制剂;样本处理时在冰上操作;
3、杂蛋白多,建议处理目的蛋白;
4、抗体特异性不强,建议使用特异性强的抗体;
5、抗体孵育时间过久,建议减少抗体孵育时间;
6、二抗与抗原有交叉反应,建议选择合适的封闭物;
7、二聚体或多聚体存在,建议增加蛋白质变性过程及强度;
8、底物显色与曝光时间过长,建议缩短显色及曝光的时间。
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