Where can I find a protocol for Co-IP for protein-protein interaction?

More Navjyoti Chakraborty's questions See All

What kind of statistical analysis should be performed?

Suppose I have a wild type and mutants A, B and AB. I take control (untreated) as well as treat each of them (i.e the WT and the mutants A,B and AB) with some treatment/s and perform RT-qPCR for...

10 November 2014 5,008 9 View

What kind of statistical analysis should be carried out for RT-qPCR data, which has been used to validate microarray data?

Suppose I have around 20 differentially expressed genes for which RT-qPCR data has been generated, and the samples are a wild-type plant and a knock-out mutant. Which statistical analysis should...

09 October 2014 2,839 17 View

What is the maximum log2fold change detectable for repressed genes using qPCR?

We need to validate our microarray data for a few down-regulated genes which show log2FC values near -7. Can qPCR detect this level of down-regulation?

03 April 2014 3,403 11 View

Can anyone suggest a tool which can take AGI/TAIR ids as input and map them onto actual pathway maps?

See above

03 April 2014 5,598 2 View

Can someone suggest the controls for GST pull down?

I am attempting GST-pull down assay for finding unknown interacting partners using GST-fusion protein. Normally bait control, prey control, resin control are taken but I am planning to take also...

31 December 2013 9,894 3 View

Has anyone used the Xcelris plant RNA isolation minikit?

I have previously used the qiagen plant RNeasy minikit and that works very well, but I would like to know about the performance of this one.

11 December 2013 1,494 2 View

Nitrate estimation in plants?

Can anyone suggest a reliable method to estimate nitrate content in plant tissue other than kjeldahl method?

04 May 2013 5,930 8 View

Method/protocol needed

Does anyone have a protocol/suggestion on leaching out all forms of N from soil or a method to determine content of nitrate, nitrite or any other form of N present in soil?

02 March 2013 4,205 0 View

Expression of membrane protein

I am trying to express a plant membrane protein using pGEX vector in BL21 strain. I could express the protein but all went into the pellet fraction. Should I purify the protein from the pellet...

01 February 2013 3,058 14 View

Can anyone tell me if it's possible to modify mbSUS (mating based split ubiquitin system) to generate and screen cDNA library?

mbSUS has been used successfully by several researchers to screen for protein-protein interaction for membrane proteins but I could not find if anyone has used it to screen cDNA library. if it is...

10 November 2012 724 1 View

Why might my GFP tag be dissociating from my plasmid after transfection?

I have a set of stably transfected cell lines all transfected with plasmids containing GFP tags on the C terminus. During a western blot using anti-GFP antibody, one of my plasmids has dissociated...

01 March 2021 9,310 4 View

Precipitate after adding laemmli buffer to protein solution containing Potassium Acetate?

Hi, I am running a size exclusion chromatography experiment with a buffer containing Potassium Acetate as a salt. I analyse these fractions through SDS-PAGE. After boiling my SEC fractions in...

01 March 2021 2,622 3 View

What is the most suitable server for predicting protein structures in bacteria?

I have used the i-Tasser several times, however it has been unavailable for several days. I tried the swiss-model, but the output was not very pleasant due to the model used. Is there any other...

28 February 2021 4,521 3 View

Do you need the CMV enhancer before the CMV promotor for a proper function of CMV on the gene that follows?

I'm a student and I have to produce a cell line with a knockin for the NRF2 gene with GFP. I have to put a promotor in front of the GFP because the gene will be too far away from the promotor of...

28 February 2021 7,127 2 View

Are protein samples in SDS compatible with ELISA?

I have used an AllPrep DNA/RNA/Protein Mini Kit QIAGEN kit to extract RNA and protein from my samples. At the end of the protein purification, I resuspend my protein in 5% SDS. Will these samples...

28 February 2021 7,370 3 View

Anyone that can help with Maxquant TMT16plex which is missing?

Hi I am very new to proteomics analysis. I have got raw files from core facility running thermofisher TMT 16plex 4 hours. I cannot find on maxquant the option TMT16Plex there is only up to 11...

25 February 2021 1,675 3 View

How to replicate the well conditions of a crystal hit with 20%(w/v) isopropanol?

I had a crystal hit in a well with 0.1M HEPEs pH 7.5, 10%(w/v) PEG 4000, and 20%(w/v) isopropanol. Why is isopropanol in w/v here if it's just solvent? How do I recreate this well condition?

25 February 2021 9,761 1 View

Problem with ligating 2.8 kb insert into 8.5 kb plasmid?

I am trying to clone 2 copies of the same mammalian gene into the pSF-CMV-Ub-Puro Ascl (contains HindIII, KpnI and NheI cut sites) plasmid. This is what the final product is supposed to look like...

24 February 2021 6,310 3 View

What urea buffer can i use for biotagged protein extraction from streptavidin beads?

Hi all, I have been doing research on biotagged LDB1 proteins in Mel cells. I purify the proteins using M280 streptavidin beads. The yield is very low so I've been trying to optimize it by...

24 February 2021 5,749 3 View

Can I use a protein-devoid condition as my negative control in ELISA?

Hello, I am running a sandwich-type ELISA where I am trying to inhibit a protein-protein interaction using small molecules. For each experiment, I determine my Z' (Z-prime) to determine the...

23 February 2021 7,429 1 View

Thirupugal Govindarajan, Ph.D.

Here you go

http://www.promega.com/~/media/files/resources/product%20guides/protein%20interaction/chap4.pdf?la=en

http://www.nature.com/nmeth/journal/v2/n6/abs/nmeth0605-475.html

Navjyoti Chakraborty

thank you for replying Mr govindarajan and mr. devendra. I have gone through the pierce and promega literature but both of them describes methods for animal cells but I am working on a plant system.