Iba1 Antibody Staining Protocol for Immunohistochemistry


Description: This antibody is expected to recognise the isoforms represented by NP_001614 and NP_116573 but not NP_004838 Please note: Although we have some very good Abreviews on mouse, some customers were receiving inconsistent results on mouse samples. We have therefore moved mouse to the predicted species and can longer guarantee it.


Primary Antibody

Name: Iba1 Antibody

Clone: Goat polyclonal

Supplier: Abcam

Catalog Number: ab5076

Dilution: 1:100 using IHC-TekTM Antibody Diluent (Cat# IW-1000 or IW-1001) to reduce background and unspecific staining and serum blocking step is NOT needed.

Incubation Time/Temp: 60 min/room temperature

Antigen Retrieval
Device: IHC-TekTM Epitope Retrieval Steamer Set (Cat# IW-1102)
Buffer/pH value: IHC-TekTM Epitope Retrieval Solution (Cat# IW-1100)
Heat/Cool Temperature: 95-100 ºC/room temperature
Heat/Cool Time: 20 minutes/20 minutes
This antibody did not work on citrate buffer based antigen retrieval solution.

Detection Methods
Standard Method: ABC Method or LSAB Method
Enhanced Method: Polymeric Methods

Chromogen Substrate
Reagent: DAB Chromogen Substrate
Incubation Time/Temperature: 1-3 minutes/room temperature

Reagent: Mayer's Hematoxylin
Staining Time: 30 seconds

Staining Pattern: Cytoplasm > cytoskeleton. Cell projection > ruffle membrane. Associated with the actin cytoskeleton at membrane ruffles and at sites of phagocytosis.
Images: Search image

Additional Information:
Species Reactivity: Rat, guinea pig, human, pig, mouse
Fixation: Formalin fixed paraffin sections
Positive Control: Brain
Negative Control: Omit primary antibody, isotype control, absorption control
Blocking: 2-5% normal serum to reduce unspecific background staining; 0.5-3% H2O2 to block endogenous peroxidase activity; avidin/biotin to block endogenous biotin activity if necessary