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