WB、IHC、ELISA
Chicken
N/A
IgY
PBS, pH 7.4, 15mg/ml Bovine Serum Albumin as Stablizer and 50% Glycerol. Store at -20°C.
Do not aliquot the antibody.
1:100-1:1,000 for immunohisto/cytochemistry
1:1,000-1:100,00 for ELLSA and Western blotting with chromogenic substrates
1:1,000-1:10,000 for Western blotting with ECL substates Optimal dilutions should be determined by the end user.
The antibody was purified from antisera by immunoaffinity chromatography using antigens coupled to agarose beads.
Antibody Specificity:Based on immunoelectrophoresis and/or EL1SA,the antibody reacts with whole molecule pig IgG.
lt also reacts with the light chains of other pig immunoglobulins. No antibody was detected against non-immunoglobulin
serum proteins.The antibody may cross-react with immunoglobulins from other species.
Store at -20°C. Avoid repeated freeze / thaw cycles.The expiration date may be extended if test results are acceptable for
the intended use.
For in vitro research use only,not for diagnostic or therapeutic use.This product is not a medical device.
1.Yongxi D ,Jun L ,Shanhui Z , et al.Design, synthesis and bioevaluation of 1,2,4-thiadiazolidine-3,5-dione derivatives as potential GSK-3β inhibitors for the treatment of Alzheimer's disease[J].Bioorganic Chemistry,2023,134106446-106446. (IF 5.1)
答:可能的原因有:
1、膜没有完全均匀湿透,建议 使用100% methanol浸透膜;
2、洗膜不充分,可以增加洗液体积和洗涤次数;
3、电极不平衡或者加样位置偏斜,可以调整电极和加样;
4、封闭物用量不足,可以提高封闭物浓度,孵育时保证封闭液完全浸没转印膜;
5、封闭物使用不当,比如检测生物素标记的蛋白时不可用脱脂奶粉封闭;
6、封闭时间不够,一般情况下室温37度封闭1小时以上或者4度封闭过夜;
7、抗体非特异性结合,可以降低抗体浓度,减少孵育时间;
8、一抗稀释度不适宜,可以对抗体进行滴度测试,选择适宜的抗体稀释度;
9、一抗孵育的温度偏高,建议4℃结合过夜;
10、抗体浓度过高或洗涤不够,建议降低抗体浓度,增加洗涤次数和时间;
11、化学显色底物过多,建议按说明书加入适量的显色底物。
答:可能的原因有:
1、胶浓度不对,不同浓度的胶跑出的蛋白条带的位置可能有所偏差,可以调整浓度;
2、抗体孵育不充分,建议增加抗体浓度,延长孵育时间;
3、酶失活,建议直接将酶和底物进行混合,如果不显色则说明酶失活了。选择在有效期内、有活性的酶联物;
4、目的蛋白存在翻译后修饰或剪切体,建议查询相关文献确定;
5、标本中不含靶蛋白或靶蛋白含量太低,建议设置阳性对照比对结果,增加标本上样量。
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