Annealing temperature in RT PCR - Positive control differes from target gene?

The primer annealing temperature , after the gradient PCR analysis, for the target gene differs from the optimal annealing temperature in housekeeping gene (positive control). What to do ? The...

06 July 2019 7,073 4 View

Fish DNA Isolation , problem with DNA degradation ?

After several protocols for gDNA isolation i have noticed that in all samples teh DNA is totally degraded. I have used several papers, some of them...

02 March 2017 9,220 7 View

RP-resolving power of primer?

08 September 2016 3,351 0 View

Why do I get UREA PAGE GEL smeared bands?

11 December 2015 8,898 14 View

Why does my diagnosis restriction digest not work when PCR verification of my insert works?

08 September 2014 2,133 5 View

Any suggestions for Brucella isolation?

I am in the middle of my project- expression of U-OMP16. I am very pressured for time, and cannot wait for a fresh sample of infected animal with brucellosis in order to isolate the Brucella...

11 December 2013 7,323 0 View

Does anyone know where I can purchase pBI101 or pBI121 plasmids and how to get Agrobacterium tumefaciens strain GV3101?

Plasmid - vectors

07 August 2013 8,921 3 View

Darwin Software - pylogenetic tree construction based on allel number ?

Respected, Does anyone know how to construct a phylogenetic tree base on (PIC) and heterozygosity (H) values calculated for each SSR primer using Darwin software ? The PIC and heterozygosity...

01 January 1970 4,481 13 View

Why does blocking with my immunogenic peptide cause nuclear fluorescence in ICC?

Hi all, I am validating a polyclonal antibody raised against human ANT4, through WB and ICC. In most of my controls, the antibody shows very little off-target binding, but when I pre-incubate the...

03 March 2021 5,999 3 View

Have someone successfully used the ER alpha (Santa Cruz sc-8002) antibody in immunostaining ?

Santa Cruz now recommends this new antibody (F-10) but I can't find a publication using this reference. I'm working on human et non human primate hypothalamus and so far, it doesn't work. Is...

02 March 2021 10,003 2 View

Issues with Permeabilization during Flow Cytometry?

Hello Everyone. Currently I am working to characterize macrophages in the myocardium after ischemia-reperfusion injury in rats. Due to the low total cell number isolated from rat hearts I can...

01 March 2021 3,867 3 View

Does anyone know an alternative product for cardiac Troponin T antibody for Immunofluorescence (Thermo # MA512960)?

We have been using Thermo Fisher anti-cardiac Troponin T antibody for immunofluorescence staining in FFPE mouse heart sections but it was recently discontinued. The literature shows it to be the...

01 March 2021 4,937 1 View

Is it possible for two human neuronal cell lines to have different gene sequence for the same protein?

I am looking at the ATP1A2 (Sodium/Potassium ATPase alpha subunit 2) in two human neuronal cell lines. Expression levels of this protein seems to be almost equal when detected by one antibody....

01 March 2021 3,607 3 View

How does an antibody have full intact antibody + half antibody?

Hello, I was just wondering how does a antibody have full intact antibody + half antibody? Is it because of the drug conjugation?

28 February 2021 1,577 1 View

Does anyone use antinuclear antibody (ANA) and anti-Phospholipid antibody ELISA kit for Non Human Primate serum samples?

I am looking for anti-nuclear antibody (ANA) and anti-Phospholipid antibody ELISA kit for Non Human Primate serum samples. Any suggestions highly appreciated. Thank you.

28 February 2021 6,512 3 View

RIA. How to measure?

When you use RIA, with a control the unknown sample with antibodies, you bare in mind the binding sites of the antibodies. However you still need to measure labelled antigen (radioactive) and not...

28 February 2021 2,133 3 View

Anyone know of a good progesterone antibody?

I am looking for a good progesterone antibody to quantify levels of progesterone in zebrafish. Applications would be ELISA, and immunohistochemistry.

28 February 2021 5,929 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