Anti-RPL7 Antibody Gives Ok Results

Biochemistry
Heidelberg University
Graduate Student

Overall

Quality of Results

Ease-of-Optimization

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Company:

Merck (Sigma-Aldrich)

Product Name:

Anti-RPL7 antibody produced in rabbit

Catalog Number:

HPA058373-100UL

Clone Number:

N/A

This antibody has been used to recognize the RPL7 protein in human cells.

Experimental Design and Results Summary

Applications

Western Blot

Sample

Hela cells, WCL

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 5% in PBS 0,05% tween 1 hour

Tertiary Incubation

None

Detection

ECL

Results Summary

Hela cells WCL has been loaded on a 4-12% SDS-precast gel (1:15h; 180 V) then proteins have been transferred on nitrocellulose membrane using the semi-dry blotting (1h 11V). Blocking with milk 5% in PBS 0,05% tween (1 hour) was followed by incubation 1h at RT with anti-Rpl9 antibody (1:3000 in milk 5% 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 5% in PBS 0,05% tween. To wash out the secondary antibody, a second wash with PBS 0,05% tween (3x 5 minutes) was performed. ECL substrate has been used to detect the signal.

DOI or PMID #

N/A

Additional Notes

N/A

Image Gallery

Summary

The Good

Once optimized works well

The Bad

The signal needs optimizations

The Bottom Line

I would look for a nicer one.

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