How do you soak your nitrocellulose membrane before transferring during a western blot?

31 December 2014 7,758 17 View

How can one calculating the total body surface area in sprague dawley rats?

I am attempting to calculate TBSA in Sprague-Dawley rats based on the paper attached. I want to use Formula 3 (Vallois' formula), but they don't describe well the measurements that need to be...

09 October 2014 4,257 2 View

Following a bradford assay, how do you know how much protein to load onto your gel in order to get visible bands?

I am attempting to do a western blot to look for differential MMP7 expression in skin biopsies. I know the expression is differentially regulated based on immunohistochemical evidence and want to...

07 August 2014 4,528 3 View

Does anyone know about how to preserve the skin samples for use in HPLC when isolating the antimicrobial linezolid from rat?

I am interested in running human skin samples through HPLC to determine linezolid concentrations. Linezolid is an antimicrobial agent of the oxazolidinone group. I am wondering if anyone has...

06 July 2014 621 0 View

Using PCR to obtain 16s rRNA amplicons from thermally injured skin?

I am attempting to use 16s rRNA primers to amplify bacterial DNA for sequencing. Does anyone have experience using pcr to obtain amplicons? I am worried my thermocycler settings are not optimized...

02 March 2014 2,811 2 View

Can anyone help with a human PDGFR-B primer for qPCR?

I am having trouble finding a suitable primer for human PDGFR-B. Has anyone successfully used a primer and if so what was the annealing temp?

02 March 2014 9,017 2 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