Application
Immunocytochemistry
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 24°C
Sample
Human Cell (Kelly Cell line)
Specification
Kelly Cell line
Permeabilization
No
Fixative
Paraformaldehyde
Other product details
Incubation time
2 hour(s) and 0 minute(s) · Temperature: 24°C · Diluent: 1% PBSA
Dilution
1/50
Secondary antibody
Name
Non-Abcam antibody was used: Goat Anti Mouse Biotinylated
Host species: Goat
Clonality: Polyclonal
Conjugation: Biotin
Host species: Goat
Clonality: Polyclonal
Conjugation: Biotin
Dilution
1/200
Additional data
Additional Notes
Human Kelly Neuroblastoma cell line was cultured in RPMI/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 PBS
Primary - tap off serum block add primary in 1% PBSA for 2h at RT
Wash 2x2min PBS
Secondary - Goat Anti Mouse Biotin 5ul/ml in 0.1% PBSA 30mins at RT
Wash 2x2min PBS
ABC 30min at RT
Wash 2x2mins PBS
DAB (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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投稿 Apr 28 2014