Histochemical localization of immunoreactive substances using labeled antibodies as reagents. | Contact experts in Immunohistochemistry to get answers
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Questions related to Immunohistochemistry
Is there such a case that certain antibodies have limited access to the intra-cellular domain of its receptor and this results in a negative staining? I have been struggling trying to detect...
07 March 2017 2,774 4 View
I'm currently slicing mouse brains that have been fixed in 4% paraformaldehyde (and embedded in agar) on the leica VT1200S. I've been having some issues with evenness of tissue in the slices and...
02 March 2017 6,149 2 View
Hi all I am having a trouble with my immunohistochemistry. I am trying to detect a secreted protein in FFPE mouse tissue sections, and I use concentration-matched goat IgG as my negative control....
01 March 2017 3,239 5 View
I've previously done multi labelling immunofluorescence experiments using 405 conjugated secondary antibodies, which worked well. This was in non perfused mice, where the brain was sliced in...
01 March 2017 5,075 2 View
I want to do a double immunofluorescence staining, and I am wondering how to choose the secondary antibody, if both of the primary antibodies are from the same host; rabbit. In other words, I...
21 February 2017 6,154 7 View
Hi everyone! I am trying to perform IHC for F4/80 (ab6640 abcam) on pfa fixed LUNG samples, but i'm not able to obtain a good staining. Do you know a good protocol? I usually use a concentration...
21 February 2017 2,736 2 View
I am looking at the expression of a particular protein in the skin by immunofluorescence and I would like to know which cells express this protein. For that, I stained both cryosections and...
21 February 2017 7,220 1 View
We have been using the anti-Human CD16 clone H27 antibody for immunohistochemistry of sections from formalin-fixed paraffin blocks. We need to re-order it but Biorad apparently no longer make...
15 February 2017 4,808 2 View
Hello, everybody. I want to perform IHC with my tissue sample. I have the antibodies for the markers that I want to see, but all of them are conjugated with the dye such as FITC, PE or APC. Can...
07 February 2017 4,768 10 View
We store our brain tissue at -20°C (usually in 50% phosphate buffer, 30% ethylene glycol, and 20% glycerol). Would NaCl in the solution interfere with cryoprotection (i.e., would it allow freezing...
24 January 2017 8,368 1 View
We have raised a rabbit antiserum against a protein coupled peptide. This peptide has been reported to be antigenic and some papers have been published with a commercial preparation raised in the...
23 January 2017 5,746 4 View
Hi there, I am doing immunohisto-staining and all my results came negative.. I washed the sections with 200 microliter of PBS. and now I am thinking that I should have immerse them in PBS rather...
18 January 2017 7,316 8 View
Hello, I am trying to optimize our Olig2 immunohistochemistry staining protocol for PFA fixed mouse brain. I found a very helpful website that reviewed several recent publications and found...
12 January 2017 4,394 6 View
After doing an alpha-MSH DAB staining in mouse PVN sections, some of them show some 'projections' thicker than normal, and also cell bodies stained. Since it is an alpha-MSH staining, I didn't...
11 January 2017 6,690 3 View
I am performing IHC using mouse brain slices. I am performing double labeling and the blocking buffer I am using is normal goat serum. One target remains constant in my experiments, and the second...
05 January 2017 5,029 3 View
Hello everyone, In order to cut 10 µm slices of mammospheres with cryostat, I had to colour them with eosin to be able to follow them while cutting.What a bad idea you'll tell me, because eosin...
20 December 2016 5,351 5 View
My antibody didn't work at all. I have mouse brains, 30 um cryosectioned, and tried different concentrations of tween, as well as both formamide and SSC treatment to no avail.
19 December 2016 6,807 3 View
In my lab we generally used to do immunocytochemistry regularly but now its not working that means it does not show any signals after secondary antibody addition with background or almost no stain...
18 December 2016 5,068 7 View
my cells contains luciferase, when i injected cells into mice, the Bioluminescence imaging showed well. but when i do the tissue immunohistochemistry, i found the luciferase express in nuclea,...
16 December 2016 2,333 3 View
Hi all! I am currently working on a project where we have gone from using mouse embryos (E7.5-9.0) to pig embryos (E.17-20). We have notices of late a lot of issues with the technical aspect of...
15 December 2016 2,758 4 View
Dear researchgaters, I have a question regarding staining protocols for oxidative tissue markers via IHC (4-HNE, MDA-2, 8-0HdG etc). I have found that these markers work in cryo stored material,...
13 December 2016 8,260 3 View
CD3, CD5 Parrafin section immunohistology. Dear Colleagues, I have also problem in Immunostaining of Parrafin sections of Rat sciatic nerve and spleen with CD3 and CD5. I am also using rabbit...
12 December 2016 5,294 3 View
Hello, I'm having some trouble with my negative control slides when I do immunofluorescence. I only work with frozen muscle tissue, and I'm trying to optimize a number of antibodies. For some of...
08 December 2016 3,523 7 View
Hi everyone, I plan to perfuse-fix my mice with 4% PFA through the heart, carefully extract the brain and then post-fix the brain overnight in 4% PFA for 12-16 hours. I plan to paraffin embed them...
17 November 2016 3,358 3 View