Should I look at BCL-2 in whole cell or subcellular fraction?

I'm looking at BCL-2 in cultured cells. I'm comparing treatments that appear to inhibit apoptosis. I'd like to look at BCL-2 levels in whole cell lysates, but I'm wondering if I should be looking...

10 November 2017 5,467 3 View

What are the differences between measuring T-cell response via ELISpot and FACS?

What is the difference between measuring IFN-y positive PBMCs with and ELISpot and using FACS to measure the % of CD8+ antigen specific T-cells? What are the functional differences between the two...

08 September 2016 6,478 0 View

What is the best way to increase signaling in an ELISpot assay?

I'd like to know if it would be better to increase the number of cells plated or increase the peptide concentration? Which one would result in a better increase in spot formation.

03 April 2016 10,037 5 View

How should I store PBMCs in serum free media that I've isolated for an ELISpot assay?

I'm isolating mouse PBMCs for an ELISpot assay. I work as quickly as possible but the cells do sit for 60-90 min while I'm counting cells and getting everything ready for the assay. The PBMC are...

02 March 2016 2,696 9 View

Why is it that x-rays with lower mean effective energies produce more scatter?

In soft tissue, why do lower kV x-rays produce more scatter and I guess have a higher RBE? I understand higher energy x-rays have more of a forward momentum, but I would think that the Compton...

02 March 2016 6,780 0 View

How can I establish good 4T1 tumors in-vivo?

I'm growing 4T1 cells in mice. I'm implanting two contralateral tumors per mouse. I'm having a hard time getting the tumors to grow symmetrically. I understand they won't all be perfect, but I'm...

05 June 2015 8,018 7 View

Protocl for measuring BCL-2 in SK-OV-3 cultured cells?

01 January 1970 8,477 2 View

How can I measure MMP-1 protein activity or protein level from liver tissue?

Instead of checking only the mRNA level, I want to check the active protein level of MMP-1 in Liver tissue from mice. How can I do that?

03 March 2021 1,763 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

IHC on mouse liver hepatocytes?

Can anyone help me choose primary antibodies for mouse liver hepatocytes and also in conjunction Cd8 cells. Also Cd3 cells. I want to look at hepatocytes and adjacent immune cells in mice with...

01 March 2021 6,953 1 View

How to avoid ulcers in s.c. injected tumors ?

Recently, we have injected s.c. B16F10 tumor cells in BL/6 mice. At around 12-15th day some animals showed ulcerations in tumors , with a big hole in the middle (it looks like a volcano). We have...

01 March 2021 9,938 3 View

Interested to stain mice tissue with WGA 488 conjugate, anybody has a protocol to recommend?

Interested to stain mice brain tissue with WGA 488 conjugate, appreciate if anyone willing to offer me some guidance or suggestions! Thank you!

28 February 2021 3,951 2 View

Looking for genomic DNA extraction kits from mouse tissue (centrifuge-based, not spin-column)?

Hello. I am looking to extract genomic DNA from mouse brain and lungs and I need a kit that is not spin-column based because I need to scale up the solution amounts for 400mg tissue (most cap at...

25 February 2021 155 2 View

What are the pros and cons for mice versus rats?

Hi Everyone, I am designing a pilot study using rodents to assess the effects of simulated space environments (radiation + altered gravity) on microbiome and immune system. I have limited...

24 February 2021 1,825 3 View

Is it harmful to breed for Cre Homozygosity initially?

Hello, I am currently using a Cas9 knock in system that will be expressed in the liver. To do so STOP codon fl/fl Cas9 KI mice were bred with Albumin-Cre mice to generate heterozgous pups, that...

24 February 2021 5,759 3 View

How to empty large intestine in mice ?

Hi all I am working on colon-targeted drug delivery for which I kept mice fasting for 6h using fasting grills during scotophase (dark period). I observed liquid fecal content in the cecum and...

23 February 2021 7,701 2 View

Does PHA-P stimulate the proliferation of lymphocytes from mice?

Hello, I will start performing MLR using lymphocytes from mice's spleen and I would like to know the better way for the stimulation. If is using PHA-P or CD3/CD28? I would like to know as well if...

23 February 2021 5,476 4 View