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

Offlavor pond fish and usage of bio-organic agent ?

Impressive sharings! Regarding on that matter,somehow people use kind of tamarind whichever in paste or liquid to get rid offlavor on flesh fish,wondering is it live fish can be done that way...

11 April 2020 2,155 3 View

How does R programming language add non numeric operators?

Say I have list x(a vector of character names) and list y(a vector of data). I want to run each element of data within y through a loop, but I want it to print the element from vector x that has...

06 April 2020 7,867 3 View

Suggestions for immunofluorescence of rat spinal cord?

I am having difficulty getting specific binding with my rat spinal cord immunofluorescence. My protocol is as follows: 1. Remove spinal cord and quick freeze in TissueTek OCT. 2. Cut tissue on...

03 September 2019 9,215 2 View

What did my western blot fail???

I am a beginner in western blot and the results were okay for the last few weeks with beta actin as housekeeping control. The transfer buffer ran out a few days ago so I prep a new 10X transfer...

13 February 2019 3,274 9 View

Where can i get the specifications for the different layers that make up tire?

here is paper i wish to replicate before i get into my thesis

03 February 2016 4,211 4 View

Any advice on the transfection reagent for mouse ES cells?

Hey guys, I have try several transfection reagent (Xfect, lipofectamine 3000, PolyJet) for transfecting mouse ES cells. But the transfection efficiency is quite low. The best I can get is 50%...

23 October 2015 5,369 9 View

Why am I getting crooked bands in SDS-PAGE?

Why am I getting this type of pattern? I am thinking the problem is either the running buffer or the sample buffer. I have changed new running buffer but the bands are still crooked. Could the...

30 January 2015 8,216 11 View

What is the reason for fungs appearing in the crystal growth system when conducting slow evaporation?

01 January 2015 5,354 3 View

How might we be able to preserve the past through old or traditional buildings?

How can people keep the past when the city is being moved forward?

25 November 2013 1,787 2 View

Gel electrophoresis result for total RNA samples, 1.5 Agarose gel was used, How to improve the integrity of RNA?

Gel electrophoresis, RNA degradation, RNA extraction from fresh tissue

02 March 2021 5,433 5 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

Problem with shRNA transfection. Why are only two plates getting contaminated instead of all?

I transfected my LNCaP-WT cells with 3 shRNA plus their NTC two weeks ago and split two puromycin selected cell plates on Friday last week(Feb 26). I checked for GFP in the cells, and they all...

28 February 2021 4,949 3 View

Any specific pipeline or parameter to analyse total transcriptomics study of brain tissue samples?

I have two groups of brain samples, control and treated for example. It was total RNA nova seq sequencing. I tried all the available pipeline like: star+rsem+deseq2, Hista+stringtie+cuffdiff,...

27 February 2021 356 6 View

Why is the efficiency of viral mRNA electroporation in PBMC low?

Hello, I have been struggling with the electroporation of PBMC with HIV-I mRNA. Although the quality of the RNA generated is good, and the handling of the cells in meticulous, the frequency of...

27 February 2021 9,233 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

What is the best concentration of cDNA to be used for qPCR?

I have synthesised cDNA from 500ng/ul of RNA. The concentration of the cDNA Is about 5000ng/ul. Should I dilute my cDNA or just use it as it is for my qPCR? I usually use 2ul of cDNA for a 20ul...

24 February 2021 8,618 3 View

Recommendations on homogenizers for RNA isolation?

I will be performing RNA isolation with frozen lung biopsy samples and am in the market for a tissue homogenizer. I am wanting a homogenizer that will fit in a 1.5ml tube. I was looking at the...

24 February 2021 3,623 3 View

RNA-seq DEGs downstream analysis and reporting in literature?

I used EdgeR to get the list of DEGs (differentially expressed genes) from my liver RNA-seq experiment. I have done the gene overrepresentation and GSEA based pathway analysis. I am currently...

23 February 2021 8,969 1 View

Is it efficient to do liquid biopsy to investigate the gene expressions of the tumor?

To do liquid biopsy and look for specific gene expressions there are so many published papers. Thera are different protocols and procedures used to isolate circulating cell free RNA or mRNA from...

23 February 2021 7,326 5 View