Immunocytochemistry abreview for Anti-68kDa Neurofilament / NF-L antibody

Good
Abreviews
Application
Immunocytochemistry
Sample
Mouse Cell (Neuro2a)
Permeabilization
No
Specification
Neuro2a
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 24°C
Fixative
Paraformaldehyde

Other product details

Incubation time
2 hour(s) and 0 minute(s) · Temperature: 24°C · Diluent: 1% PBSA
Dilution
1/500

Secondary antibody

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

Additional data

Additional Notes
Mouse Neuro2a cell line was grown on cover slips in 6 well plates in DMEM/10% FCS. Cells were washed with PBS prior to fixing with 1%PFA. 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 PBS Primary - tap off serum block add primary in 1% PBSA for 2h at RT Wash 2x2min PBS Secondary - Goat Anti Chicken 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 Apr 22 2014

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