What are differences in anti-fade mounting media in IHC/ICC?

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Why are specific proteins or genes only expressed in the specific body part?

My question is as so many proteins/genes are present in a cell or a tissue how and why specific ones are only expressed?

04 May 2014 2,853 1 View

PBST or TBST for western blot?

What is good for western blot PBST or TBST? Sometimes people use only TBS and PBS also, and still results good with no background.

09 October 2013 392 7 View

RIPA Buffer in protein lysis?

Generally I use RIPA buffer for protein lysis for western Blot from tissues and cells. But I have noticed it interferes while doing estimation of proteins by Bradford's method. However I also...

07 August 2013 1,875 3 View

How can I solve a problem in maintaining H9C2 cells?

I'm having a problem in maintaining H9C2 cells for a long duration. After 2-3 splits cells become unhealthy as black particles start coming. Also sometimes this problem happens from the beginning....

07 August 2013 4,578 4 View

What are the various techniques to see how much your protein gets phosphorylated?

I want to see the how much my protein gets phosphorylated. But its phospho-antibody is not available in the market. Is there any method except doing western blot with phospho antibody of same target?

07 August 2013 5,110 2 View

Can I change concentration and brand of antibodies mid-experiment?

I am doing immunohistochemistry of the c-Fos protein and I have a budget problem. For context, the experiment has been done over 3 years prior to my arrival and I have 40 brains done and 100 left....

29 July 2024 5,961 2 View

Sodium azide in immunohistochemistry?

Hello everyone, A collaborator sent us his protocol in order to be able to use his immunohistochemistry kits. But one step contains PBS + 0.3% H2O2 + 0.1% sodium azide. Knowing the dangerousness...

23 July 2024 6,314 3 View

How to label synapses in over-fixed mice brain sections (40 um) via immunohistochemistry (IHC)?

We have mice brains that were over-fixed due to old PFA used during perfusion. Thus, the synapses are no longer being labeled by the Synaptophysin (SY38) mAB. which works perfectly every other...

17 July 2024 7,767 3 View

Is perfusion required to stain brain tissue with every method?

I'm looking to immunostain some brain tissue collected previously. The animal was *not* perfused prior to tissue collection. Will the presence of blood interfere with all staining methods...

14 July 2024 177 4 View

Can I use BSA for a Histological Blocking Solution?

Hi all, I would like to know if it is possible to use Bovine Serum Albumin (BSA) in my blocking solution for immunofluorescence muscle fiber typing. The primary antibodies' host is mouse. The...

07 July 2024 7,967 3 View

To what degree does the immunohistochemical (IHC) evaluation of Ki-67 hold diagnostic value beyond neoplastic pathologies?

In the context of doctoral research, I utilized immunohistochemical (IHC) analysis to quantify the expression levels of the Ki-67 proliferation marker. While the application of this test and...

25 June 2024 7,825 2 View

How I can select appropriate level to adjust clustering effect?

I have multi-stage data. The ICC is different at all level. Say at stage 1 =0.1, at stage=0.2 and at stage 3=0.3. I want to run melogit regression. Please, which variable should be in...

16 June 2024 6,265 4 View

ImageJ Concentric Circles?

Hi there! We are trying to use the Concentric Circles Plugin on ImageJ to determine signal intensity in neurosphere cryosections after immunocytochemistry/confocal analysis. We determined the...

12 June 2024 7,332 0 View

Help with mounting meningeal tissues with Prolong Diamond?

Hello! I've recently been trying to optimise immunofluorescent staining of mouse meningeal tissue, and I'm having trouble mounting it just because it's so thick. We use prolong to mount and...

05 June 2024 624 2 View

Different results from western blot and icc?

hi have you ever experienced of coming different results from western blot and icc? I treated toxin (a-syn PFF) to control ES-derived midbrain doparminergic neuron and experimental group that I...

25 May 2024 3,162 0 View