Cathelicidin Antibody Staining Protocol for Immunohistochemistry

Description: Cathelicidins are a family of antimicrobial proteins predominantly found in the peroxidase negative granules of neutrophils. The cathelicidins are synthesized as preproproteins. Within neutrophils, they are stored in granules as inactive proforms after removal of the signal peptide. The active biologic domains of the cathelicidins generally reside in the C terminus. The C terminal antimicrobial peptides are activated when cleaved from the proforms of the cathelicidins by serine proteases from azurophil granules.

Primary Antibody

Name: Rabbit Anti-Cathelicidin Antibody

Clone: Rabbit polyclonal

Supplier: Abcam

Catalog Number: ab64892

Dilution: 1:500 using IHC-Tek^TM 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-Tek^TM Epitope Retrieval Steamer Set (Cat# IW-1102)

Buffer/pH value: IHC-Tek^TM Epitope Retrieval Solution (Cat# IW-1100)

Heat/Cool Temperature: 95-100 ºC/room temperature

Heat/Cool Time: 20 minutes/20 minutes

Detection Methods

Standard Method: ABC Method or LSAB Method

Enhanced Method: Polymeric Methods

Chromogen Substrate

Reagent: DAB

Incubation Time/Temperature: 1-3 minutes/room temperature

Counterstain

Reagent: Mayer's Hematoxylin

Staining Time: 30 seconds

Results:

Staining Pattern: Cytoplasmic/secreted

Images: Search image

Additional Information:

Species Reactivity: Human

Fixation: Formalin fixed paraffin sections

Positive Control: Skin

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. if IHC-Tek^TM Antibody Diluent (Cat# IW-1000 or IW-1001) is used, there is no serum blocking step needed.

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