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Guinea pig antibody to Catalase

$260USD 

Code OSC00328W

eLab Book

Osenses Protocols: Western Blot

 

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1. Run the SDS-PAGE at 250V.

 

2. Soak the gel for 10 min in transfer buffer then rinse it with Di water.

 

3. Soak the membrane for 5 min in Di water.

 

4. Semi-dry transfer at 10 V.

5. After transfer, rinse once the membrane with PBS and then wash 3x, 5 min each with PBS in 100 ml. Shake at 50 RPM.

 

6. Block with 1xPBS containing 1% LFDM for 30 min at RT with shake at 50 RPM.

 

7. Rinse with 70 ml of PBS.

 

8. Incubate with antibody O/N in the fridge (3-5C).

 

9. Wash the membrane 5 times (5 min each) with PBS-T containing 0.1% Tween-20. Wash in 100 ml with shake 50 RPM.

 

10. Add Osenses' rabbit anti Gp adaptobody at 1:3000 dilution and incubate 30 min, wash and incubate with Sigma HRP-Goat anti Rb diluted 1: 15,000). Add 6 ml per membrane (6x8 cm). Incubate 30 min to 1 hour at RT with 50 RPM shake.

 

11. Wash 4 times (5min each) with shake 50 RPM with 100 ml of PBS-T containing 0.1% Tween-20.

 

12. Wash 6 times more with 100 ml of PBS with shake 50 RPM.

 

13. Prepare the ECL substrate from BioRad.

 

14. Scan the blot on C-Digit at High resolution (12 minutes).

 

 

   
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