VEGF A - AB-90040

Immunohistochemistry Mouse - VEGFA

Experiment
Immunohistochemistry Mouse - VEGFA
Product
VEGF A - AB-90040 from Immunological Sciences
Manufacturer
Immunological Sciences

Protocol tips

Publication protocol

WT and NG2KO mice at postnatal day 6 (P6), and adult naïve and EAE-affected WT and NG2KO mice at 20 and 40 dpi, were transcardially perfused with 100–150 ml of 2% paraformaldehyde (PFA) and 0.2% glutaraldehyde PBS solution, under deep anaesthesia (ketamine/xylazine cocktail, 90 mg and 4.5 mg/kg, respectively) by intraperitoneal injection. Whole brains were removed and post-fixed by immersion in the same fixative at 4°C for 4 h, washed in PBS overnight at 4°C, and stored in 0.02% PFA in PBS at 4°C. Using a vibrating microtome (Leica Microsystem), serial sagittal sections (30/35-μm thick) evenly spaced at 200 μm intervals, were cut from each hemisphere to allow the analysis of the entire antero-posterior extension of the cerebral cortex. The sections, in a range of 60 to 120 sections/hemisphere, were stored in a multiwell archive as free-floating sections in PBS at 4°C. The archived sections were submitted to immunostaining to ascertain the presence of demyelinating lesions, and the adjacent sections were then utilized for confocal morphometry (see for details [6]). Briefly, after permeabilization (0.5% Triton X-100 in PBS), free-floating sections were incubated with single or combined primary antibodies, overnight at 4°C, and with appropriate secondary antibodies (Table 1), for 45 min at RT, and then counterstained with TO-PRO-3 diluted 1:10k in PBS (Invitrogen). Finally, the sections were collected on Vectabond treated slides (Vector) and coverslipped with Vectashield (Vector). Negative controls were prepared by omitting the primary antibodies and mismatching the secondary antibodies. Sections were examined under Leica TCS SP5 confocal laser scanning microscope (Leica Microsystems) using a sequential scan procedure. Confocal images were taken at 0.35 μm intervals through the z-, x-, and y-axes of the section, with 40x and 63x oil lenses.

Full paper   Login or join for free to view the full paper.

Reviews

VEGF A - AB-90040 from Immunological Sciences has not yet been reviewed for this experiment

We'd love it if you would be the first to write a review!

Discussion

Start your discussion

Share your thoughts or question with experts in your field

Start a discussion

Papers

Check out relevant papers found by Labettor's AI that are relevant for performing Immunohistochemistry Mouse - VEGFA using VEGF A - AB-90040 from Immunological Sciences.

View full paper   Login or join for free to view the full paper.

Manufacturer protocol

Download the product protocol from Immunological Sciences for VEGF A - AB-90040 below.

We haven't found the manufacturer protocol for this product yet.

Videos

Check out videos that might be relevant for performing Immunohistochemistry Mouse - VEGFA using VEGF A - AB-90040 from Immunological Sciences. Please note that these videos are representative and steps or experiment specific processes must be kept in mind to expect desired results.

We haven't found any additional videos for this experiment / product combination yet.

Outsource your experiment

Fill out your contact details and receive price quotes in your Inbox

  Outsource experiment
Become shareholder Discussions About us Contact Privacy Terms