Immunocytochemistry/ Immunofluorescence abreview for Anti-Iba1 antibody

Excellent
Abreviews
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (Macrophage cell line Mono-Mac 6)
Specification
Macrophage cell line Mono-Mac 6
Fixative
Paraformaldehyde
Permeabilization
No
Blocking step
Serum as blocking agent for 20 minute(s) · Concentration: 10% · Temperature: 24°C

Other product details

Dilution
1/500
Incubation time
16 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: PBS

Secondary antibody

Name
Non-Abcam antibody was used: Invitrogen Rabbit Anti Goat AF555
Host species: Rabbit
Clonality: Polyclonal
Conjugation: Alexa Fluor® 555
Dilution
1/1000

Additional data

Additional Notes
Macrophage cell line Mono-Mac6 was grown in RPMI/10% FCS and grown on glass cover slips in 6 well plates.
Fixed in 1%PFA prior to staining.
Wash 3x3min in PBS.
Block 10% rabbit serum for 20min at RT.
Primary Ab diluted 1/500 in PBS and incubated overnight at 4deg, also works by incubating for 2 hours at RT.
Wash 3x3min in PBS.
Secondary - rabbit anti-goat AF555 (red) 1/1000 in PBS for 1h at RT.
Wash 3x3min in PBS
Counterstained with Hoechst 33258 (blue) for 10mins at RT.
Wash 3min in dH2O
Coverslips removed tapped dry and placed on glass slides and mounted in Invitrogen ProLong gold and kept at 4deg until imaged.

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Submitted May 25 2012

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