Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (Spinal Cord (Dorsal Horn))
Antigen retrieval step
None
Permeabilization
Yes - 1% Triton, 0.25% Tween in 1XPBS
Specification
Spinal Cord (Dorsal Horn)
Blocking step
Mouse-on-Mouse Block (Vector Labs) as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 25°C
Fixative
Paraformaldehyde
Other product details
Incubation time
1 hour(s) and 0 minute(s) · Temperature: 25°C · Diluent: Mouse-on-Mouse Diluent (Vector Labs)
Dilution
1/200
Secondary antibody
Dilution
1/500
Name
Non-Abcam antibody was used: Goat anti Mouse IgG (H+L) Alexa Fluor 594
Host species: Goat
Clonality: Polyclonal
Conjugation: Alexa Fluor® 594
Host species: Goat
Clonality: Polyclonal
Conjugation: Alexa Fluor® 594
Additional data
Additional Notes
ab18184 was utilized to detect the 6XHis tag epitope conjugated to a phage display antibody against heparin sulfate. We observed ab18184 binding both in the presence and absence of the phage display antibody, suggesting that ab18184 binds an endogenous epitope in the mouse spinal cord. We were thus unsuccessful in utilizing this antibody for IHC-P applications in mouse CNS tissue.
Abcam response
The possibility of false staining is due to secondary being binging to endogenous mouse. The mouse on mouse (MOM) kit we sell is a best product we offer to block the endogenous IgGs. Reviewing the details we believe the endogenous IgGs may be source of the problem.
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.
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投稿 Feb 11 2016