Immunocytochemistry abreview for Anti-Aquaporin 4 antibody

Good
Abreviews
Application
Immunocytochemistry
Sample
Rat Cultured Cells (33B)
Specification
33B
Fixative
Paraformaldehyde
Permeabilization
No
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 24°C

Other product details

Dilution
1/100
Incubation time
2 hour(s) and 0 minute(s) · Temperature: 24°C · Diluent: 1% PBSA

Secondary antibody

Name
Non-Abcam antibody was used: Vector Goat Anti Rabbit Biotinylated
Host species: Goat
Clonality: Polyclonal
Conjugation: Biotin
Dilution
1/200

Additional data

Additional Notes
33B oligodendroglioma cell line was cultured in DMEM/10% FCS on coverslips in 12 well plates.
Cells were washed with PBS and fixed in 4% PFA until staining.
Staining protocol:
Wash 2x2min PBS
Endogenous peroxidase block - 3%H2O2 in methanol for 15mins at RT
Wash 2x2min PBS
Serum block - 10% Goat serum in 1% PBSA
Primary - tap off serum block add primary in 1% PBSA for 2h at RT
Wash 2x2min PBS
Secondary - Goat Anti Rabbit Biotin (Vector) 5ul/ml in 0.1% PBSA 30mins at RT
Wash 2x2min PBS
ABC from Vector 30min at RT
Wash 2x2mins PBS
DAB (Vector) (brown)
Wash 2x2min H2O
Counterstain - Harris' Haematoxylin (purple)
Wash off with H2O
Dehydrate 2mins - 1x75%EtOh 2x100%EtOh 2xHistoclear
Mount using Histomount

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Submitted Jul 06 2012

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