Immunoblotting

Immunologic method used for detecting or quantifying immunoreactive substances. The substance is identified by first immobilizing it by blotting... | Contact experts in Immunoblotting to get answers

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Questions related to Immunoblotting

Immunoblotting CD63 in exosomal proteins?

I'm having trouble in immunoblotting(western) CD63 especially in exosomal proteins, since the antibody works well in non reducing conditions. My proteins were in RLN buffer. Is this going to...

03 February 2014 406 4 View

Anyone have experience with immunoblot for watercrystal damaged samples?

If watercrystal damage happened in my whole brain samples, would there be any significant differences in the result of my immunoblotting between step A and step B (which are as follows)? Step A:...

02 February 2014 906 2 View

Anyone know the implications of Imaging Nitrocellulose Blot Upside-Down with Seemingly Intact Signals?

I am wondering the implications of imaging western blots that have been vertically inverted (i.e. lanes still run 1-15 from left to right, but columns run from lightest to heaviest from top to...

02 February 2014 7,187 3 View

Immunoblotting CD63 in exosomal proteins?

I'm having trouble in immunoblotting(western) CD63 especially in exosomal proteins, since the antibody works well in non reducing conditions. My proteins were in RLN buffer. Is this going to...

02 February 2014 7,640 5 View

How to study the DNA repair efficacy of a cell?

I am trying to study a DNA damage repair efficacy of cultured mammalian cancer cells following drug treatment. I have already checked the DNA damaging property of the drug through COMET assay and...

15 January 2014 9,801 11 View

How to study the DNA repair efficacy of a cell?

I am trying to study a DNA damage repair efficacy of cultured mammalian cancer cells following drug treatment. I have already checked the DNA damaging property of the drug through COMET assay and...

01 January 2014 6,522 12 View

Which subcellular membrane markers should be chosen to test purity of plasma membrane by immunoblotting? where can I buy them?

I want to evaluate the purity of plasma membrane-enriched fractions isolated in our laboratory by immunoblotting with antibodies raised against known subcellular membrane (endoplasmic reticulum,...

18 December 2013 3,866 2 View

Which subcellular membrane markers should be chosen to test purity of plasma membrane by immunoblotting? where can I buy them?

I want to evaluate the purity of plasma membrane-enriched fractions isolated in our laboratory by immunoblotting with antibodies raised against known subcellular membrane (endoplasmic reticulum,...

12 December 2013 3,138 2 View

What are the possible problems of mass spectrometry(MS) from a one-dimensional (1D) gel?

I made 1D-western blotting of different crude antigens of Schistosoma mansoni. Now I need to perform the MS. I have read about the subject and I would like to know if I have to do the 2D-gel...

12 December 2013 7,867 7 View

Recommended polyclonal ab for detection of mCherry?

Looking for a good polyclonal anti-RFP antibody with low background for mCherry detection. If you have a link to the paper you used it in so much the better!

12 December 2013 9,921 1 View

Do you have any suggestions regarding Odyssey infrared immunoblotting?

Blocking buffers (Licor, Rockland) are expensive (up to 200€ per 500 mL). Is there any less costly alternative out there?

10 October 2013 2,659 9 View

Blank western blots: is my secondary antibody contaminated/degraded?

I've been using western blots for about a half a year successfully, but recently my blots began resulting in completely blank signal. I'm confident my problem is not resulting from transfer issues...

10 October 2013 2,775 9 View

Can I use an IB antibody for IP?

Can I denature the antigen source, such as cell lysate (RIPA), and then use the IB (immunoblotting) antibody as IP (immunoprecipitation) antibody to precipitate target protein from cell lysate?

27 August 2013 8,830 15 View

Type I collagen Western Blot?

I would like to show type I collagen alpa2 (COL1A2) downregulation on protein level. Immunoblotting using 5 different antibodies didn't yield any bands at all (not even positive controls)....

17 August 2013 8,385 16 View

Can I use an IB antibody for IP?

Can I denature the antigen source, such as cell lysate (RIPA), and then use the IB (immunoblotting) antibody as IP (immunoprecipitation) antibody to precipitate target protein from cell lysate?

08 August 2013 4,475 16 View

Type I collagen Western Blot?

I would like to show type I collagen alpa2 (COL1A2) downregulation on protein level. Immunoblotting using 5 different antibodies didn't yield any bands at all (not even positive controls)....

08 August 2013 4,670 16 View

What are the best transfer conditions for membrane proteins like the EGF receptor?

I have been having a difficult time with my EGFR western blots (175kD membrane protein). It looks as though the EGFR is not transferring well or it is not binding the membrane. I have had it...

07 July 2013 6,406 24 View

Is there any chemical (acid/base hydrolysis) or else that can be used to degrade LPS?

I just want to see the difference between LPS bands in bacterial cells and degraded LPS bands on an immunoblot.

07 July 2013 9,035 0 View

Rabbit primary antibody used in western blot

Has anyone used rabbit primary antibody for western blot? I am going to try to blot my protein of interest using a western blot kit, namely Western Max tm HRP chromogenic detection kit.The kit is...

04 April 2013 3,982 15 View

Is casein tyrosine-phosphorilated?

I am currently doing some western blotting analysis to identify some specifically regulated phospho-proteins. I am using 4G10 platinum antibody (millipore) to recognize tyrosine-phosphorylations....

03 March 2013 715 3 View

Problem with developing.

I find that when I apply the ECL solution to my membrane, develop it in the dark room and put it back into the western container, if I try to develop again (reapply ECL solution, place into...

03 March 2013 6,804 13 View

I'm having problems with Immunoblot (chemiluminescence revelation) with Anti Col1A1

So, we're having problems with immunoblot with AntiCol1A1. We're having a big background and we can barely see some bands we have already made with the same samples and, despite having some...

02 February 2013 2,125 2 View

I am doing some work on E.coli and wanted to find out if anyone knows of an immunoblot against type 1 fimbriae?

Our research project is evaluating the effects of chinese herbs on the treatment of urinary tract infections. We are now looking at specific effects of herbs on the fimbriation process.

29 January 2013 1,022 1 View

I am doing some work on E.coli and wanted to find out if anyone knows of an immunoblot against type 1 fimbriae?

Our research project is evaluating the effects of chinese herbs on the treatment of urinary tract infections. We are now looking at specific effects of herbs on the fimbriation process.

01 January 2013 9,600 1 View

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