WB, IHC, IF
H, R, M, Mk, Dg, Ch, Hm, Rb, Pg, Sh
43KD
IgG1
PBS, pH 7.4, containing 0.02% sodium azide as Preservative and 50% Glycerol. Store at -20°C.
Do not aliquot the antibody.
WB: 1:5,000
IHC: 1:200
IF: 1:200
Antibody can detects endogenous β-actin protein.
β-Actin is one of six different actin isoforms that have been identified. The actin molecules found in cells of various species
and tissues tend to be very similar in their immunological and physical properties. Therefore, Antibodies against β-Actin are
useful as loading controls for Western Blotting. However it should be noted that levels of β-Actin may not be stable in certain
cells. For example, expression of β-Actin in adipose tissue is very low and therefore β-Actin should not be used as loading
control for these tissues.
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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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