Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (pancreatic cancer)
Specification
pancreatic cancer
Fixative
Acetone+Methanol 1:1
Permeabilization
Yes - PBST
Blocking step
Vector Lab MOM kit #BMK-2202 as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: RT°C
Other product details
Dilution
1/100
Incubation time
4 hour(s) and 0 minute(s) · Temperature: 20°C · Diluent: PBS
Secondary antibody
Name
Non-Abcam antibody was used: Alexa 488 Goat anit-Mouse
Host species: Goat
Clonality: Polyclonal
Conjugation: Alexa Fluor® 488
Host species: Goat
Clonality: Polyclonal
Conjugation: Alexa Fluor® 488
Dilution
1/1000
Additional data
Additional Notes
In order to avoid Mouse on Mouse artifact and background MOM kit (Vector Lab, #BMK-2202) was used. Autofluorescence in such relatively weak staining is an issue. Aldehyde fixatives (PFA) react with proteins (amines) to generate fluorescent products the same is with Glutaraldehyde. I have avoided aldehyde-induced autofluorescence it in this case (but that's general advice) by using fixture of acetone+methanol 1:1 in -20degC for 10min. This mixture both fixed and permabalized the 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.
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提交于 Oct 04 2010