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Pippuri
Member since: Jul 12, 2009
From: Georgia, United States
Status: Protein Chemistry & Glycobiology Moderator
My points: 997    what's this
Name: [Privacy]
 


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Jun 6, 2013    02:31 PM

Replied to topic in Proteomics forum   Co-Immunoprecipitation Kit

Hi Sue, Now, the real issue that you are encountering is- whether your antibody is suitable for IP? Have you tried to address this issue before proceeding to the co-IP experiment? ie. performing an IP with your antibody and then immunoblotting to detect your protein of interested.

Jun 3, 2013    10:01 AM

Replied to topic in Proteomics forum   Co-Immunoprecipitation Kit

Hi Sue, If you really want a co-IP kit, there is a selection to choose from. (that I know others have used successfully) I never brought a kit for co-IP. If you have done IP before, co-Ip is just to "tune-down" your detergents in the lysis and wash buffers, otherwise the steps are t ...

Jun 3, 2013    09:55 AM

Replied to topic in Protein Detection (Western blots, gels, IP) forum   No Bands in Western Blot

Hi Anjali, How much protein you loaded per lane? (You did mention 100ug of protein was mixed with sample buffer, you didn't mention how much you used per lane) While primary antibodies are typically stored in an azide contining buffer, it is not necessary to remove azide proir to using them i ...

Apr 27, 2013    01:01 PM

Replied to topic in Protein Detection (Western blots, gels, IP) forum   Multiple Antibodies on Western Blot?

Hi ScienceGeek, Yes, as long as these proteins are well separated on the SDS-PAGE. Good luck.

Apr 27, 2013    07:54 AM

Replied to topic in DNA (PCR, Real Time PCR) forum   TE Buffer after DNA extraction

Hi cpd, it is perfectly okay to store DNA in the dry form. Leave them in -20 deg instead of 4 deg just to be safe. Good luck.

Apr 19, 2013    06:32 AM

Replied to topic in Cytology, Histology and IHC forum   Correlation of mRNA and protein

Hi sitara, The levels of mRNA and protein do not necessary to go hand-in-hand. It can go through a feed back mechanism per your postulation or via other (more complicated) regulatory networks. If the protein level stays the same, one question to ask it that, how long the protein half-life i ...

Feb 28, 2013    05:42 PM

Replied to topic in Protein Detection (Western blots, gels, IP) forum   M2 beads non specific binding

Hi CHAMBOO, It sounds like you can try to use less M2 beads. Using too much beads often pulls down non-specific proteins! Good luck.

Feb 28, 2013    05:40 PM

Replied to topic in Protein Detection (Western blots, gels, IP) forum   Purification of the cytosolic protein

Hi Sompop, please consult the following website- http://www.agrisera.com/cgi-bin/ibutik/AIR_ibutik.pl?funk=steg_tva&artgrp=36 Good luck

Feb 28, 2013    05:38 PM

Replied to topic in Protein Detection (Western blots, gels, IP) forum   Stronger signal after stripping my western blot

Hi research_BY, Did you get a new bottle of secondary antibody or open a new box of ECL during that the time that you started to have problem with your westerns? It sounds like that conditions of your westerns are not optimized.... The "weak signal" that you were getting before str ...

Feb 28, 2013    05:30 PM

Replied to topic in Protein Chemistry forum   Size exclusion chromatography troubleshoot

Hi Cheriyan, What is the Mw of the "contaminated" species you are trying to get rid of?

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