Apolipoprotein D Antibody Staining Protocol for Immunohistochemistry


Description: Apolipoprotein D is a glycoprotein involved in the human plasma lipid transport system. It is mainly associated with high density lipoprotein particles and forms disulphide-linked heterodimers with other apolipoproteins. The function of apolipoprotein D in the metabolism of plasma lipoproteins is unclear but the observation that this protein forms complexes with lecithin: cholesterol acyltransferase has led to the suggestion that apolipoprotein D may be involved in cholesterol esterification and transport of substrates and products of the reaction. Apolipoprotein D is expressed in a range of normal tissues including axillary apocrine glands, adrenal cortex and corpus luteum. Peripheral nerves, pituitary, testis, cerebellum and renal tubes are also positive.


Primary Antibody

Name: Apolipoprotein D Antibody

Clone: 36C6, Mouse Anti-Human

Supplier: Novocastra Labs

Catalog Number: NCL-Apo-D

Dilution: 1:40 - 1:80 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: 60min/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

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

Reagent: Mayer's Hematoxylin
Staining Time: 30 seconds

Staining Pattern: Granular staining of Apolipoprotein D-containing lipid droplets
Images: Search image

Additional Information:
Tissue Type: Testis, Prostate carcinoma tissue lysate.
Fixation: Formalin-fixed paraffin sections.
Positive Control: Testis, Prostate carcinoma tissue lysate.
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