Membrane anchored protein?

Hi, Is there any way to prove that a protein is membrane anchored/peripheral membrane protein? Any simple trick or any technique will be helpful. Thanks

24 November 2019 1,008 5 View

E.Coli expression strain for expressing mycobacterial membrane protein gene?

Hi, I have cloned some three different constructs which are putative membrane protein of Mycobacterium tuberculosis into pET28a with N-term His tag. I tried to express them in BL21, Rosetta, C41,...

17 December 2018 8,536 2 View

Rosetta (DE3) plasmid throw out?

Hi, I have rosetta (DE3) with incorporation of tRNAs for AGG, AGA, AUA, CUA, CCC, GGA codons on a compatible chloramphenicol-resistant plasmid. I cloned a gene in kanamycin resistant...

30 July 2018 6,527 2 View

Cleaning of Supherose 6 Increase 30/100 GL?

Hi, I feel that my Supherose 6 column is contaminated with some protein because I get pretty clean SDS gel bands after Ni NTA but after SEC I see multiple bands on SDS PAGE. How should I clean...

10 April 2018 1,200 3 View

NO plasmid after isolation?

Hi, I am cloning a 2.8 kb gene into pet28b vector. I did two ligation reaction. In one of them, I did not put insert (only vector and ligase) (control reaction). Then I transformed 10 microlitre...

15 February 2018 1,784 4 View

Is writing a paper a good way to introduce an idea or concept?

Is writing a paper a good way to introduce an idea or concept to be considered by the greater public, industry, etc? I am wondering if sharing an idea through an academic journal may be good way...

01 January 1970 5,527 8 View

No title

01 January 1970 1,741 1 View

Can someone suggest a way to prepare slides to observe under confocal microscope for observing fluorescent tagged proteins??

I am growing the cells on coverslips and transfecting the fluorescent tagged protein directly on them. Then i want to observe these cells under confocal microscope. Currently i am just using...

01 March 2021 6,142 3 View

Will a 10xHis-tag prevent anti-MBP from binding to MBP?

I have a protein with a 10xHis-MBP tag on the N-terminus. I'm fairly certain I am expressing my protein, but I'm not getting a signal on my western blot. Is there a possibility that the 10xHis tag...

25 February 2021 8,404 3 View

How to minimise the risk of being abused the profile of potential participants tagged in a social media post?

Dear colleagues, I got this one in one project I did ethics review. It's great to hear about your thoughts/experiences. "Doing an open recruitment online (i.e., via Facebook) requires public...

22 February 2021 3,322 3 View

Unable to detect a truncated protein variant, but I can observe a truncated cDNA from Sanger?

Hi, so basically... I am trying to mimic a clinically observed condition wherein a frame-shift mutation happens only at one allele of the patients, giving rise to a truncated variant which evades...

17 February 2021 1,130 6 View

Choice of Ab for Co-IP: Does an anti-(tag) antibody (such as anti-flag/ anti-myc) a must if I am using co-transfected cells for Co-IP?

Hi guys, I am trying to detect the binding of 2 proteins by Co-immunoprecipitation. However, the choice of antibody is confusing me. I tried probing the endogenous protein right away but the...

16 February 2021 9,050 5 View

Can anyone suggest me broad host range fluorescence expression vectors for Gram-positive bacteria ?

Hi everyone, I am looking for fluorescence expression vectors for expression in gram-positive bacteria present in the skin microbiome. I need to tag them with a fluorescence marker and observe...

06 February 2021 2,566 4 View

FLAG purification, elution doesn't have protein?

I am purifying a heterotetramer, with one subunit having a 1D4 tag and the other having a FLAG tag. I purified from Sf9 with HBS pH7.3 and 0.002%LMNG first with Anti-1D4 resin and got an adequate...

02 February 2021 3,860 1 View

Antibody dilution for IHQ in OCT frozen tissue is more similar to IHC in Paraffin-embedded samples or to Immunocytochemistry?

31 January 2021 8,678 3 View

How to show a protein is being dimerized in SDS-PAGE gel?

A protein dimerizes through CTD. I have expressed CTD of protein with two different tags. Now, I want to show that these CTD of protein dimerize in a pull-down assay. But my doubt is that if the...

29 January 2021 7,529 4 View

How to solubilize protein?

Hello, I have cloned my gene of interest in a bacterial expression system i.e. BL 21 Gold (DE3) and checked for protein production and solubility. Good amount of protein was observed on SDS-PAGE...

15 January 2021 7,063 5 View