Yes you can transfer a 6kDa protein with nitrocellulose membrane (0.2um). I perform western blot assays with a protein at 10kDa and highly recommend using the Life Technologies Bolt® 4-12% Bis-Tris Plus Gel, 10 well gel running system and then the Life Technologies XCell II™ Blot Module system for the transfer of protein. Just ensure when running the protein gel that the gel markers are very well separated. Hope this helps.

I agree with Anita Pinar. I would only add the recommendation of using MOPS buffer to run the 4-12% gels. You get a better resolution at the 6 KDa area of the gel.

We have done it repeatedly with adrenomedullin, another 6KDa protein. You can see our published protocols:

Martínez A, Miller MJ, Unsworth EJ, Siegfried JM, Cuttitta F. Expression of adrenomedullin in normal human lung and in pulmonary tumors. Endocrinology 136:4099-4105 (1995).

Montuenga LM, Martínez A, Miller MJ, Unsworth EJ, Cuttitta F. Expression of adrenomedullin and its receptor during embryogenesis suggests autocrine or paracrine modes of action. Endocrinology 138: 440-451 (1997).

Serrano J, Fernández AP, Sánchez J, Rodrigo J, Martínez A. Adrenomedullin expression is up-regulated by acute hypobaric hypoxia in the cerebral cortex of the adult rat. Brain Pathology, 18:434-442 (2008).

Sackett DL, Ozbun L, Zudaire E, Wessner L, Chirgwin JM, Cuttitta F, Martínez A.

Intracellular proadrenomedullin-derived peptides decorate the microtubules and contribute to cytoskeleton function. Endocrinology, 149:2888-2898 (2008).

Thank you all. I have read some paper and all of them recommended to coat the membranes with gelatin before transfer and fix with formaldehyde after that. Furthermore, various authors recommend to use 15-20 gels. I'll try all procedures. Thank you!

Best regards

;)

Dear Dewald, I just tried this one, in addition I did not transfer with SDS. Thank you ;)

Dear Felipe,

Were you able to detect your protein? I'm dealing also with a 6kDa protein, but I didn't have any good result.

If you succed, would you let me know which protocol do you follow?

Thanks in advance!

Yazmine