c-Myc antibody (D84C12) Rabbit mAB

Biochemistry
Heidelberg University
Graduate Student

Overall

Quality of Results

Ease-of-Optimization

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Company:

Cell Signaling Technology (CST)

Product Name:

c-Myc

Catalog Number:

5605

Clone Number:

D84C12

Good antibody for the detection of c-Myc and very sensitive to c-Myc expressions changes

Experimental Design and Results Summary

Applications

Western Blot

Sample

Hela cells, cell lysate

Primary Incubation

1:1000 milk 1% in PBS 0,05% tween over night

Blocking Agent

Milk 5% in PBS 0,05% tween

Secondary Incubation

Rabbit 1:3000 milk 1% in PBS 0,05% tween 1 hour

Tertiary Incubation

None.

Detection

ECL

Results Summary

This antibody is very good and specific. Hela cells WCL has been loaded on a 4-12% SDS-precast gel (1:30h; 180 V) then proteins have been transferred on nitrocellulose membrane using the semi-dry blotting (1h 12V). Blocking with milk 5% in PBS 0,05% tween (45 minutes) was followed by incubation overnight at 4°C degree withc-Myc antibody (1:1000 in milk 1% in PBS 0,05% tween) Then the membrane was washed with 3x 5 minutes with PBS 0,05% tween, followed by a 1-hour incubation with Rabbit secondary antibody 1:3000 in milk 1% in PBS 0,05% tween. The secondary antibody was washed out with a second wash with PBS 0,05% tween (3x 5 minutes). ECL substrate has been used to detect the signal.

DOI or PMID #

N/A

Additional Notes

N/A

Related Categories

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Summary

The Good

Easy and fast to use

The Bad

It can give a bit of background sometimes, playing with the milk percentage and PBS tween washes is reccomended

The Bottom Line

Highly suggest buying it

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