Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Rat Tissue sections (Growth plate cartilage)
Antigen retrieval step
Enzymatic - Buffer/Enzyme Used: 700U/ml hyaluronidase
Permeabilization
Yes - 0.2% Triton X 100 in TBS
Specification
Growth plate cartilage
Blocking step
10% goat serum + 1% BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: RT°C
Fixative
Formaldehyde
Other product details
Incubation time
16 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: 10% goat serum + 0.1% Tween 20
Dilution
1/1000
Secondary antibody
Dilution
1/1000
Name
Abcam antibody:Goat Anti-Mouse IgM mu chain (HRP) preadsorbed
Additional data
Additional Notes
Performed on 8 week rat tibia.
-Dehydrate slides per usual
-Enzymatic treatment in hyaluronidase 600U/ml 37deg 1hr
-Rinse 3x5min TBS
-Permeabilize 0.2% Triton X 100 in PBS 15mins RT
-Rinse 3x5min TBS
-Hydrogen peroxide quench in 3% H2O2 in methanol 15mins RT
-Rinse 3x5 min TBS
-Block 10% goat serum + 1% BSA 1hr RT
-Primary incubation 1:1000 overnight 4deg
-Rinse 3x5min TBS
-Secondary 1:1000 ab98679 1hr RT
-Rinse 3x5min TBS
-Develop with DAB 2 mins
-Hematoxylin counterstain
-Rehydrate per usual
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.
MR. Chris Erickson
Verified customer
提交于 Jan 06 2016