Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (white adipose)
Specification
white adipose
Fixative
Formaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citrate, 10mM pH6
Permeabilization
Yes - 0.4% Triton X-100
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
Other product details
Dilution
1/250
Incubation time
2 hour(s) and 0 minute(s) · Diluent: Blocking buffer (or overnight at +4oC)
Secondary antibody
Name
Non-Abcam antibody was used: AlexaFluor anti-rabbit-488
Host species: Chicken
Clonality: Polyclonal
Conjugation: Alexa Fluor® 488
Host species: Chicken
Clonality: Polyclonal
Conjugation: Alexa Fluor® 488
Dilution
1/500
Additional data
Additional Notes
This antibody works well in Western blotting (1:1000-3000), with PVDF. Also with IP (1:300-500 used, very high efficiency of pull down, 0.5mg total protein) - both using murine WAT lysates using 1% Triton X-100 in PBS and protease/phosphatase inhibitors (see figure). Also works in immunocytochemistry on various cell types and frozen brain sections (both 4% PFA fixed) when using at 1:500 and fluorescent secondary antibodies at 1:1000 using standard protocols. It does not appear to work at any concentration when staining formalin-fixed adipose section in paraffin, using fluorescent detection, nor with antigen retrieval (Citrate, as attached). Did not have any more success when using DAB as a substrate with ABC system and biotinylated secondary antibodies.
Abcam response
Many thanks for this feedback; we invite you to submit separate Abreviews for ICC and IHC brain and adipose tissue sections.
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.
Abcam user community
Verified customer
投稿 Feb 25 2013