WB, IP, IF
N/A
N/A
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
IP: 1:200
IF: 1:1000
The Flag tag antibody can recognize C-terminal, internal, and N-terminal Flag-tag fusion proteins.
The DYKDDDDK peptide (Flag-tag) is a small component of an epitope which does not appear to interfere with the bioactivity
or the biodistribution of the recombinant protein. It has been used extensively as a general epitope tag in expression vectors.
It can be used for affinity chromatography, then used to separate recombinant, overexpressed protein from wild-type protein
expressed by the host organism. It can also be used in the isolation of protein complexes with multiple subunits.A Flag-tag can
be used in many different assays that require recognition by an antibody. If there is no antibody against the studied protein,
adding a Flag-tag to this protein allows one to follow the protein with an antibody against the Flag sequence.
更新中...
答:可能的原因有:
1、目的蛋白有多個修飾位點,本身可以呈現多條帶,建議查閱文獻或進行生物信息學分析,獲得蛋白序列的修飾位點信息,通過去修飾確定蛋白實際大小;
2、樣本處理過程中目的蛋白發生降解,建議加入蛋白酶抑制劑;樣本處理時在冰上操作;
3、雜蛋白多,建議處理目的蛋白;
4、抗體特異性不強,建議使用特異性強的抗體;
5、抗體孵育時間過久,建議減少抗體孵育時間;
6、二抗與抗原有交叉反應,建議選擇合適的封閉物;
7、二聚體或多聚體存在,建議增加蛋白質變性過程及強度;
8、底物顯色與曝光時間過長,建議縮短顯色及曝光的時間。
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