Gel purification of linearized plasmid, gave low yield why?

I am using Qiagen Gel extraction kit for gel purification of linearized plasmid. The procedure is followed as per the kit. if i load 1ug DNA in agarose gel, i am getting 600ng DNA or less after...

16 October 2020 6,667 4 View

Dear researchers, In RE digestion with Nco I i am getting smearing. Can any one please advice?

well1- 2 log ladder well2- uncut plasmid(3.2kb) well3- cut wit NcoI well4-cut wit NcoI tried with less time of incubation, low amount of enzyme doesnot work

10 September 2020 6,711 3 View

How to find the list of journals with "Your Paper Your Way"?

03 May 2020 2,492 5 View

Ideal way to administer the powder diet into rat?

I need to feed the rat with powder high fat diet. We used to mix the powder high fat diet with coconut oil and make the powder diet into small balls. But, the rats are not eating these balls...

16 November 2016 6,341 5 View

How to compare the groups in which each group consists the data for two different duration?

03 November 2016 6,237 1 View

How do you confirm the solubility apart from inspection?

22 February 2015 2,299 3 View

Which triton (x 100 or wr1339) might be used to induce hyperlipidemia?

In order to screen the hypolipidemic drugs, Triton induced hyperlipidemic animal model is being used. In most of the papers, Triton WR 1339 (tyloxapal) was used to induce hyperlipidemia. However,...

24 January 2015 2,111 3 View

Does any one experience with ideal diet induced hyperlipidemic animal model?

We need to test our herbal drug with diet induced hyperlipidemic animal model (Sprague Dawley rats) and we have gone through literature to know about high fat diet. There are many protocols...

25 December 2014 8,858 2 View

Which is the best real time PCR detection system among BioRad, Applied Biosystem and Roche?

We are going to buy a real time PCR instrument for our laboratory and we received a quotation from BioRad, Applied Biosystem and Roche companies. The price of all three makes is feasible with our...

09 October 2014 3,571 15 View

Which is the best method for preparing plant extracts with polar, non-polar and mid-polar solvents?

I understand that there are several methods available to prepare plant extract namely maceration, decoction, infusion, Soxhlet Extraction, Microwave assisted extraction (MAE), supercritical fluid...

01 October 2014 7,894 8 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