Which forward primer I should order?

I have been trying to design primers for microRNA expression. The image shows that there are 2 sets of forward primer (Tm calculated in two different manners) and one universal "reverse primer" as...

10 November 2018 5,216 0 View

How can I characterise HMEC cells?

I have a set of breast cancer cell lines and will use HMEC as a control population. Since HMECs have the tendency to show unusual characteristics along with the passages so I would like to know...

07 August 2018 6,616 0 View

Pros and cons regarding expression profiling of microRNA-SNP using breast cancer cell lines?

I have found a SNP in a microRNA gene from my association study and now want to do the expression analysis in breast cancer cell lines (sub-type specific). What are the factors should I consider...

04 May 2018 674 0 View

How can I design vector with miRNA variant (SNP) for expression analysis?

I have found a miR-SNP which is significantly associated with my breast cancer case-control cohort. Now I want to see its expression in different breast cancer cell lines to find out the best...

03 April 2018 5,145 4 View

How can I view .map & .ped files in command promt in Windows using PLINK?

I have got .bed .bim and .fam files from a database and need to see whats inside. But I don't know the command to do so. As well as I would like to view my own .ped and .map files in the command...

08 September 2017 8,758 0 View

In vitro assay to screen anti-arthritic property of drug compound?

09 October 2016 9,263 4 View

Which cell-cycle phase is the best for drug treatment?

I have used the drug as a broad term here. Specifically my focus is on those drugs which have antioxidant property. Is there really any time point at which cells will respond best to the...

07 August 2016 4,857 0 View

HUVEC starvation, time and with or without serum?

To get the best ROS generation (i am expecting 40-50% increase than that of control), how long my HUVEC should be starved and what would be the serum percentage? i am using EBM-2 (Lonza) without...

06 July 2016 9,227 5 View

HBSS, with or without Ca2+/Mg2+ for DCFDA assay?

06 July 2016 1,167 5 View

Why HUVEC is giving low amount of protein?

Trying to measure protein concentration for Lucigenin assay. But even culturing at more than 80% confluency, after centrifugation and sonication getting only 330ug/ml of protein. Hard to maintain...

05 June 2016 1,227 2 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

What's the concentration recommend for recombinant protein use in western blotting?

I'm currently working having problem with the concentration for recombinant protein use in western blot, the one been used is Recombinant Human CD19 Fc Chimera Protein-rndsystem which does not...

24 February 2021 8,514 3 View

What is the best way to manage Western blots?

Increasingly, more articles are being retracted because of duplicate or otherwise dubious Western blots. Consequently, journals are asking for original Western blots and authentication of...

23 February 2021 8,609 3 View

How to manage western blot for proteomics data validation?

Hi Everyone, I am a bigginer in western blotting and I am trying to use it for validation of my proteomics data. I have: two regulated proteins with molecular mass of 40 and 73 kDa one control...

23 February 2021 6,671 4 View

Diltuions for a Western blot?

I need help using the equation C1V1=C2V2 figuring out how much antibody to add to use for a western blot. The stock was origianlly 3.45 ng/ul but diluted 1:10 so now its 0.345 ng/ul. I need to run...

22 February 2021 9,901 3 View

Can anyone help me how to get the similar bands in Actin?

I run western blot on HCT-116 cell lines, I got actin band like in the attached picture. The band size is not same. Can anyone help me explain this situation and how to overcome? Thank you your...

22 February 2021 6,937 3 View

I see 2 random bands in my ERK/pERK blot? Has anyone else seen this?

I ran a western blot using 12% gel and probing for ERK/pERK (cell signaling antibodies). The green is total ERK, the Red is supposed to be phospho-ERK. I see 2 bands below the total ERK that...

22 February 2021 1,777 3 View

Will SARS-CoV-2 Spike be broken into S1 and S2 by denaturing conditions?

Hello, I am performing a western blot of my SARS-CoV-2 Spike-pseudotyped lentiviral particles. I denature my virus by adding concentrated virus to 2X Laemmli buffer at 1:1, then boiling for...

17 February 2021 4,844 4 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

(western blot) can methanol damage my antibody signal?

I accidentally rinsed with methanol after adding my primary antibody to my membrane. I noticed after ~10 seconds and immediately replaced it with PBST (3x15 min washes). Will my antibody signal be...

12 February 2021 5,824 3 View