Re: [Histonet] IHC DAB

Hi Kristy, As the others said, so many things that could be on the way...But making sure your system is based on a HRP enzyme is definitely a good start, as mentioned by Joe. If you can share details, I might be able to help too. Welcome to the IHC world! Cheers, Ana Ana Maluenda Rese

Re: [Histonet] Pregnant in histo lab. Am I safe?

Hi Valerie, I don't know how it is in US, but in Australia what I did was to liaise with our Occupational Health and Safety Department to check which measurements they'd recommend about having pregnant workers around the lab. My lab however is not heavily Histology and the tissue processing is

Re: [Histonet] IHC mouse optomization

Hi Blanca, It all depends on what you are trying to stain, how the tissue has been harvested/processed and what are your antibodies. If you are trying to use a primary mouse on mouse tissue, that is always tricky. You will need to probably explore your options with M.O.M kits. Otherwise, you ma

[Histonet] Platelet staining on FFPE mouse tissues

Hi everyone, Does anyone has experience performing IHC staining for platelets on mouse FFPE? I've been trying for a while with different primary antibodies unsuccessfully. I found two that work in cryosections, but not a single hint of positivity on FFPE tissues. The samples have been prepared

Re: [Histonet] Unstained slides

Hi all, It has been very interesting reading all your comments on unstained slides. This has been a forever discussion I always have everywhere I go in different research institutes. So expanding the topic, I wonder what's everyone's opinion on unstained cryosections? How long are they reliable

Re: [Histonet] IHC validation

Hi Terri, This is actually a very valid note. This has been in my mind for years, but never had the chance to put in action in a research environment (I always try to bring to the research field the efficiency and money saving strategies we use in diagnostics). Where do you usually order your

[Histonet] Frozen section - IHC for 6x HisTag

Hi everyone! Does anyone have experience with immunohistochemistry or immunofluorescence anti-6x HisTag on frozen sections, mouse tissue? I can find online lots of references for immunocytochemistry, but not much on tissue staining. Any help or hints would be much appreciated! Kind regards, A

Re: [Histonet] Paraformaldehyde Fixed Tissue

Hi Sandra, I haven't myself particularly worked with brain and spinal cord, but majority of my protocols in my old job used fixation in 4% PFA (24 hours at 4-8oC) and routinely process (or transfer to graded EtOH, if not processing immediately). However, our routine process didn't include a firs

[Histonet] IHC for secreted proteins and cytokines in frozen sections - Pre-fix or post-fix?

Hi everyone, Just wondering what is people's opinion and protocols for IHC in mouse frozen sections targeting secreted proteins and cytokines. I see lots of places using fresh frozen sections/snap-freeze and cold acetone or methanol/ethanol post-fixation. Is this an issue when it comes to diffu

[Histonet] H&E on frozen sections

Hi everyone, Just wondering if anyone out there had already had issues with commercial Haematoxylin and Eosin. I started working in a lab that recently purchased Sigma Mayer's Haematoxylin (MHS-16) and 1% Alcoholic Eosin (Fronine - II016). We work with fresh frozen sections of mouse tissue. Sin

[Histonet] Anti-fibrin antibody

Dear all, I've been trying to optimise an immunohistochemistry protocol for fibrin that has not been working. Was wondering if there is anyone that would have any recommendation of staining protocols for fibrin or recommendation of anti-fibrin antibodies? We are working with cryosections, mouse