[Histonet] Luxol Fast Blue with IHC

Hello- Has anyone tried to counterstain an IHC stain with Luxol Fast Blue? I am worried that the LFB destain will remove some of the IHC. I use the VIP solution from Vector as the chromogen for IHC instead of DAB and I destain Luxol Fast Blue with Hydroquinone/Sodium Sulfite (in addition to

[Histonet] Heterochromatin Ab

Hello- I'm wondering if anyone can recommend a good antibody that specifically labels heterochromatin (IF on FFPE sections). I've looked into several histone markers and HP1 (although I'm unsure which subtype would be best), but I'd prefer to use an antibody that I know has worked for someone

[Histonet] Frozen Artifacts

Hello Histonet... I am using frozen brain sections and I have terrible frozen artifacts. I receive the tissue from a brain bank so I have no control over the method of freezing. Is there ANYTHING that I can do at this point to at least reduce these artifacts? I've tried formalin fixation

[Histonet] Cresyl Violet Counterstain of LFB

Hi Histonet... Quick question!! I'm doing Luxol Fast Blue staining on frozen human brain sections (1% LFB in 95% EtOH with Hydroquinone/Sodium Sulfite Differentiator) and trying to counterstain with Cresyl Violet. Following Cresyl Violet incubation (0.1% for 5') my protocol calls for me to

[Histonet] Hypoxia in Post-Mortem Samples

Hey there! I have some post-mortem brain samples (FFPE) with varying PMI's and I'd like to determine the hypoxia levels (and location) at time of death (to identify if this may play a role in the disease I am studying). First of all, does anyone have any suggestions as to a good hypoxia

[Histonet] Autoflourescence

Hi- I am having a horrible time with autofluorescence in my human brain FFPE tissue. I have been using Sudan Black which helps a little, but the background is still pretty bad. The tissue is usually cut at 10-15um. Does anyone have any other suggestions? Has anyone tried photobleaching the

[Histonet] Myelin Stains for LCM

Hi- I am trying to find a suitable Myelin stain for LCM that maintains RNA integrity. I am currently testing Luxol Fast Blue, Oil Red O and Fluoromyelin but none seem to keep the RNA in tact all that well (although I am still trying to modify these procedures). My ultimate goal is to dissect

[Histonet] Primary works with ABC but not IF

Hello- I am in a situation where my primary antibody works using the ABC method, but I do not see staining with immunofluorescence. My IF protocol works well with other antibodies so I do not know why it's not working with this particular antibody. Does anybody have any suggestions? I have

[Histonet] Polyclonal Proteolipid Protein Ab?

I have been (unsuccessfully) trying to find a polyclonal antibody against human proteolipid protein (stains myelin) that will work with immunofluorescence on FFPE sections. I have also tried some myelin basic protein with no luck. Has anyone found any in the past that have worked?? Thanks!

[Histonet] citrisolve?

Does anyone have any experience using Citrisolve as a Xylene alternative to deparaffinize tissue sections? Do I use it just like Xylene for the same amount of time? Can it be reused or should I use new Citrisolve each time? How do I dispose of it? Any help at all would be much appreciated.

[Histonet] Antigen Retrieval

Hi- I am new to using IHC and I need some advice on Antigen Retrieval. I usually do it in an autoclave for 15min at 121C for FFPE human brain tissue (in Citrate or Tris Buffer), but by the end my tissue looks pretty bad and parts have even fallen off. The tissue is fixed on SuperFrost Plus