western blot antibody customer review for anti-fyn polyclonal antibody (stj93171)
Post on 14-Feb-2017
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Cell Line:
Rat brain tissue lysate, mouse brain tissue lysate, rabbit IgG(55KD). Jurkat, HL-60, K562, Raji & 6T-CEM human whole
cell lysates.
Method of validation: Western Blot
Primary Antibody: STJ93171 Anti-Fyn antibody
Secondary Antibody HRP goat anti-rabbit
Dilution ratio: 1:1000
Protocol
Gel electrophoresis: 5% polyacrylamide gel, constant voltage 55 V.
Transfer: 0.45um NC membrane, 1 piece of gel (wet transfer), constant current 150 mA, 70 minutes.
Blocking: TBS with 5% skimmed no-fat milk for 1.5 hours on shaker.
Primary antibody probing: primary antibody was added to the solution of TBS with 5% skimmed no-fat milk , 4C̊ for overnight on shaker.
Membrane wash: TBST wash 3 times for 5 minutes each.
Secondary antibody probing: secondary antibody was added to the solution of TBS with 5% skimmed no-fat milk with 1:3000 dilution ratio, 4C̊ for 2 hours on shaker.
Membrane wash: TBST wash 3 times, for 10-20 minutes each.
Visualization: ECL chemiluminescent visualization
Antibody Customer Review: STJ93171 Anti-Fyn antibody Antibody Rating:
Gel electrophoresis information:5% polyacrylamide gel, constant voltage 55 V.
Transfer information:0.45um NC membrane, 1 piece of gel (wet transfer), constant current 150 mA, 70 minutes.
Lane No. Antigen Loading
amount
Primary anti-
body
Primary anti-
body dilution
ratio
Secondary
antibody dilution
ratio
Target band KD
1 Rat brain lysate
50 ug
STJ93171 Anti-
Fyn antibody 1:1000 1:3000
~60KD
2 Rat brain lysate
3 Mouse brain
lysate
4 Mouse brain
lysate
5 Rabbit IgG (55KD)
6 Jurkat lysate
7 HL-60 lysate
8 K562 lysate
9 Raji lysate
10 6T-CEM lysate
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