EDTA Buffer (1mM EDTA, 0.05% Tween 20, pH 8.0):
EDTA (Sigma, Cat# E-5134) ----------- 0.37 g
Distilled water -------------------------- 1000 ml
Mix to dissolve. Adjust pH to 8.0 using 1N NaOH. Then add 0.5 ml of Tween 20 and mix well. Store this solution at room temperature for 3 months or at 4 C for longer storage.
Note: This buffer works excellent for many antibodies, but it often gives high background staining (maybe due to endogenous biotin revealed after this pretreatment). So primary antibody can often be highly diluted. It is very useful for low affinity antibodies or when tissue antigens are not intense.
EDTA 缓冲液 (1mM EDTA, 0.05% Tween 20, pH 8.0):
EDTA (Sigma, Cat# E-5134) ----------- 0.37 g
Distilled water -------------------------- 1000 ml
搅拌至溶解. 用1当量的氢氧化钠溶液调节到pH to 8.0。然后加 0.5 ml 的Tween 20 再混匀。EDTA 缓冲液在室温可以贮存三个月，在摄氏四度可以长期贮存。
注: 这个缓冲液对用于许多抗体非常好，但时常会由强背景染色。所以一抗可能要多稀释一些。EDTA 缓冲液非常适用于亲和力比较低的一抗或组织抗原不强的情况。
