Phospho-ERK1/2 detection in HL-1 via Western blotting

October 30, 2014

Kli.Pha.-Tox
Charité - Universitatsmedizin Berlin, CCM
Ph.D.

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Phospho-ERK1/2 protein identified using Phospho-p44/42 MAPK (Erk1) (Tyr204)/(Erk2) (Tyr187) Mouse mAb.

Company:

Cell Signaling Technology

Product Name:

Phospho-p44/42 MAPK (Erk1) (Tyr204)/(Erk2) (Tyr187) Mouse mAb

Catalog Number:

5726S

Phospho-ERK1/2 protein expression was characterized via western blot in HL-1 cells.

Experimental Design and Results Summary

Applications

Western blot

Sample

Cell lysate of murine HL-1 cells

Primary Incubation

1:500; overnight , 4°C , in TBS-T; 1%BSA; 0.02 Na3

Blocking Agent

5% low-fat milk solution

Secondary Incubation

1:5000, 1 h, room temperature, rabbit-anti-mouse IgG – HRP conjugated

Tertiary Incubation

5% low-fat milk solution

Detection

ECL Plus Detection Reagents from Amersham

Results Summary

Using the above mentioned antibody (1:500) 3 bands were detected including one for phospho-ERK1/2 at 44 kDa.

Additional Notes

None

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Phospho-ERK1/2 protein identified using Phospho-p44/42 MAPK (Erk1) (Tyr204)/(Erk2) (Tyr187) Mouse mAb.

Summary

The Good

Three bands were detected including one for phospho-ERK1/2 at 44 kDa.

The Bad

None

The Bottom Line

Using the above mentioned antibody (1:500), 3 bands were detected including one for phospho-ERK1/2 at 44 kDa. The antibody worked well in Western blot.

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