Immunoprecipitation with nfkb?

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Crispr-cas9 knock out with Mouse cell line?

Hi, anyone have suggestions for crispr cleavage detection using thermo genomic cleavage detection kit? I am trying to make nfkb1 knock out in RAW 264.7 cell line and I have using thermo Cas9...

11 December 2018 4,235 0 View

Get-in fellowship?

hi, anyone can suggest some lab in us working on crisp cell line knock outs?

08 September 2018 8,147 0 View

Gene and protein name nomenclature?

Hi, all.I need to know about gene and protein name nomenclature? Basically as per rules we need to write protein name in capital letters and gene name first letter capitals and rest in small...

08 September 2018 330 0 View

Crispr-cas9 knock out with Mouse cell line?

Hi, anyone has information regarding peoples working on crispr- cas9 in mouse cell lines? I want to apply for GET-in fellowship, so I need to contact person's working on same platform in which I...

04 May 2018 269 1 View

Inos detection after H.pylori treatment in RAW 264.7 macrophages?

Hi all, I need to can we measure i-NOS in h.pylori treated samples collected in PBS? After infection of any cell line , we usually washed cells with PBS before collection of cell pellet for RNA...

04 May 2018 4,005 2 View

Cell counting for time kinetics Experiment?

Hi, I need to know about cell counting for time point experiments- I am working on Raw 264.7 cells which has 11 hours doubling time. I use to seed 1 Million cells in six well plate which has...

31 December 2017 697 3 View

H.pylori 26695 wild type strain?

HI, all I am working on H.pylori strain 26695 since 2 years but now its showing nay increase in il-beta or Inos production when infecting raw cells with same. this is bacteria we can borrowed from...

31 December 2017 5,660 1 View

Transfection protocol for raw cells?

Can anyone has standardized protocol for Transfection in raw cell line? I have Thermo Ltx reagent? Is it vary with plasmid type? I have Pcdna 3with flag and his tag?? please suggest plasmid conc....

31 December 2017 1,267 5 View

License for knock out permit?

Hi, All Can anyone tell me is JNU has license to import knock out mice?

11 December 2017 593 0 View

Preparation of cell line knock out using crisper?

Hi, All, I like to know can anyone tell me or suggest me some labs in India preferably Delhi, who have done knock out in mammalaina cells using crisper technology.

09 October 2017 8,702 0 View

What precautions should be taken while handling S. aureus enterotoxin Type B in the lab?

I would like to understand potential safety concerns while handling SEB in the lab. Especially while working in animal house facility. Would like to know precautions for handling. Sigma MSDS...

07 August 2024 6,034 3 View

How to isolate lymphocytes from mouse spleen?

I have tried several times to isolate lymphocytes from mouse spleen, but all attempts have been unsuccessful. I tried most available protocols. I used different dissociation media (HBSS with Ca...

04 August 2024 9,913 7 View

How can I calculate the SOD enzymatic activities using the spectrophotometer method?

I have to quantify the enzymatic activities of SOD in plants using a method that employs hypoxanthine-xanthine oxidase to create O2. The colorimetric reaction was performed by combining...

23 July 2024 8,492 3 View

What are the strategies to Enhance IgG-Producing Plasma Cells in mice for Monoclonal Antibody Development?

I have immunized BalB/C mice with a protein using the intradermal (ID) method with Complete Freund's Adjuvant (CFA) and Incomplete Freund's Adjuvant (IFA), following a 14-day interval and three...

22 July 2024 9,160 2 View

Promotor observation in region annotation of RNA RIP-seq?

Hello all, I extracted RNA from my samples and performed RIP-seq. After annotating the genomic regions using R, I obtained promoters, exons, introns, and UTRs. Given that my samples consist of...

18 July 2024 1,579 2 View

What will be the best way to test NFkb activation via western blot?

Hi, I have lysate from human macrophages, and I wonder what will be the best way to check the NFKB activation in those cells. I was thinking p-p65 or p-ikkb./ Is that make sense?

18 July 2024 5,214 4 View

Unexpected 35 kDa Band Observed During Expression and Purification of Dengue 3 NS5 RdRp Domain ?

Background: I am developing an RdRp assay for Dengue virus-3 and need to express and isolate the NS5 protein's RdRp domain (~101 kDa). My expression vector includes a GST tag followed by a 6x His...

13 July 2024 9,950 3 View

Validated HPLC method for the estimation of the Amphotericin B ?

During the release test of Amphotericin B using 1% Tween 80, i observed three peaks even in the standard. The first peak consistently yielded results around 500%, while the other peaks showed...

08 July 2024 1,330 0 View

On what basis you choose Mass spec %Cov(95) cutoff score to validate a protein through CoIP?

I am working on centrosomal proteins. I overexpressed my POI and then I did SNAP capture pulldown followed by mass spec and I got approximately 700 protein hits. I am trying to validate specific...

03 July 2024 5,701 0 View

THP-1 cells completely dying with PMA, help?

Have been trying to induce THP-1 using PMA at different concentrations (range from 5 to 50 ng/mL) but after 48h all the conditions except the control are mostly dead, checked by trypan blue and...

03 July 2024 3,884 5 View