抗体稀释液

Antibody Dilution Buffers Primary Antibody Dilution Buffer: 1%BSA (stabilizer and blocking) 0.1% cold fish skin gelatin (blocking) 0.05% sodium azide (preservative)

0.01M PBS pH7.2 (TBS pH7.6 used in primary antibody dilution buffer produces weaker staining) Note: 1) Antibodies diluted using this buffer can be stored at 4 ºC for 6 months without reducing binding activity.

2) This buffer can not be used for diluting HRP conjugated antibodies since sodium azide is an inhibitor of HRP.

HRP-conjugated Primary Antibody Dilution Buffer: 1%BSA (stabilizer)

0.1% cold fish skin gelatin (blocking) 0.05% Thimerosal (preservative) 0.01M PBS pH7.2

Note: Antibodies diluted using this buffer can be stored at 4 ºC for 6 months without reducing binding activity.

Secondary Antibody Dilution Buffer: 0.01M PBS, pH 7.2

0.05% sodium azide (preservative)

Note: 1) Antibodies diluted using this buffer can be stored at 4 ºC for 6 months without reducing binding activity. 2) Do not use BSA or other serum containing reagents to dilute secondary antibodies since they may bind to BSA or serum therefore reducing antibody affinity. 3) Using TBS to dilute secondary antibodies often produces weaker staining. So use TBS only for the antibodies with high background staining or for alkaline phosphatase conjugated antibodies since phosphate is a inhibitor of alkaline phosphatase.

HRP-Streptavidin Dilution Buffer: 0.01M PBS, pH 7.2

0.05% thimerosal (preservative)

Note: Antibodies diluted using this buffer can be stored at 4 ºC for 6 months without reducing binding activity.

AP-Streptavidin Dilution Buffer: 0.05M TBS, pH 7.6

0.05% thimerosal (preservative)

Note: 1) Antibodies diluted using this buffer can be stored at 4 ºC for 6 months without reducing binding activity. 2) PBS can not be used to dilute Streptavidin-AP since phosphate inhibit alkaline phosphatase activity.

Fluorescent Dye (Avidin-FITC, Avidin-Texas Red, Avidin-AMCA) Dilution Buffer: 0.01M PBS, pH 7.2

0.05% thimerosal (preservative)

一抗稀释剂:

1 % 牛血清白蛋白 0.1% 冷海鱼皮胶 0.05% 叠氮钠

0.01M 磷酸缓冲液(PBS) pH7.2 (注: 1升)

注备: 1) 一抗稀释剂可以储存在4 ºC , 6 个月.

2) 此稀释剂不能用来稀释辣根过氧化酶 (HRP)标记的一抗,因为里面的叠氮钠会抑制辣根过氧化酶 (HRP) 的活性.

辣根过氧化酶 (HRP)标记的一抗一抗稀释剂: 1 % 牛血清白蛋白 0.1% 冷海鱼皮胶

0.05% 硫柳汞 (preservative)

0.01M 磷酸缓冲液(PBS) pH7.2 (注: 1升) 注备: 此稀释剂可以储存在4 ºC , 6 个月.

二抗稀释剂:

0.05% 叠氮钠

0.01M 磷酸缓冲液(PBS) pH7.2 (注: 1升)

注备: 1) 二抗稀释剂可以储存在4 ºC , 6 个月.

2) 不要用含有牛血清白蛋白或其它血清的稀释剂来稀释二抗,因为二抗可能会与牛血清白蛋白或其它血清反应.

3)用TBS稀释二抗会降低染色, 因此TBS用于有强背景的抗体活碱性磷酸酶标记得二抗,因为磷酸盐可以抑制碱性磷酸酶的活性.

辣根过氧化酶-链霉亲和素(HRP-Streptavidin Dilution Buffer) 稀释剂: 0.01M 磷酸缓冲液(PBS) pH7.2 (注: 1升)

0.05% 硫柳汞

注备:此稀释剂可以储存在4 ºC , 6 个月.

碱性磷酸酶-链霉亲和素(AP-Streptavidin Dilution Buffer) 稀释剂:

0.05M Tris缓冲液 (TBS), pH 7.6

0.05% 硫柳汞

注备: 1) 此稀释剂可以储存在4 ºC , 6 个月.

2) 磷酸缓冲液不能用来稀释碱性磷酸酶-链霉亲和素,因为磷酸盐回抑制碱性磷酸酶活性.

荧光染料 (Avidin-FITC, Avidin-Texas Red, Avidin-AMCA) 稀释剂: 0.01M 磷酸缓冲液(PBS) pH7.2 (注: 1升)

0.05% 硫柳汞