Cytokine immunofluorescence for tissue slices?

Hello, I am planning to do a cytokine immunoflourescence in spleen tissue slices. In bibliography most methods do not permeabilize cells, thereby detecting extracellular protein (associated to...

05 June 2018 9,441 3 View

POVA(323-339) induced cytokine profile in DO11.10 mice?

06 July 2017 6,898 1 View

Enzymatic reaction scale-down?

Hi, I'm currently working with an enzymatic reaction. The reagents are quite expensive, so I wonder if I can scale down the reaction volume from 100ul to 25ul. Is this possible or there is any...

05 June 2017 3,642 6 View

Mucins and negative-charged proteins staining?

02 March 2017 3,914 3 View

Paraformaldehyde fixation conditions for intracellular staining?

Hello, I've been comparing intracellular staining protocols and noticed that there are several alternatives for fixation with paraformaldehyde, varying time and concentration. Also, I read that...

11 December 2016 586 5 View

IL-4 staining problem in Th2 polarized cells?

Hi, I'm having trouble trying to stain mouse IL-4 from Th2 cells polarized in vitro.I've been purifying CD4+ cells from BABL/c spleens with magnetic beads and culturing them in CD3/CD28 pre-coated...

10 November 2016 8,558 8 View

How to precipitate proteins from a guanidine 8M solution?

09 October 2016 2,612 4 View

Avidin-Agarose resin high background?

Hello, I've been having trouble with biotynilated protein purification using Avidin-Agarose resin. The idea is to use the purified biotynilated molecules as samples for MALDI-TOF-MS.To check the...

06 July 2016 2,861 6 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