Application
Immunohistochemistry free floating
Sample
Mouse Tissue sections (brain)
Specification
brain
Other product details
Incubation time
24 hour(s) and 0 minute(s)
Dilution
1/300
Secondary antibody
Secondary antibody
None used
Additional data
Additional Notes
Mice were perfused with 2% paraformaldehyde in PBS and postfixed overnight in 2% paraformaldehyde. Brains kept at 4 degrees in PBS until sectioning on a vibratome (60 um). Sections stored at -20 degrees in antifreeze until IHC reaction. Antigen retrieval with 1% sodium borohydride for 20 min. Pre-incubation in 10% normal goat serum, 1% BSA, 0.3% TritonX-100 in PBS for 1 hr. Primary incubation in 1% normal goat serum, 1% BSA, 0.3% TritonX-100, plus primary antibody (1:300) in PBS overnight at room temp. Rinse 3 x 10 min in PBS. Secondary incubation in same solution as primary (but with different antibody) for 90 min at room temp. Rinse 3 x 10 min in PBS. Mount with Vectashield with DAPI.
This antibody gave bright, low-background, specific labeling of the GFP reporter, which was too dim to see on its own. In the image, green is GFP, red is NeuN, and blue is DAPI.
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.
James Bogenpohl
Verified customer
提交于 Jul 16 2018