APAF1 Antibody Staining Protocol for Immunohistochemistry

 

Description: Apoptotic peptidase activating factor 1, also known as APAF1, is a protein which in humans is encoded by the APAF1 gene. This gene is mapped to chromosome 12q23. It encodes a cytoplasmic protein that initiates apoptosis. And it is an essential downstream effector of p53-mediated apoptosis. This protein contains several copies of the WD40 repeat domain, a caspase recruitment domain (CARD), and an ATPase domain (NB-ARC). In the presence of cytochrome c and dATP, APAF1 assembles into an oligomeric apoptosome, which is responsible for activation of procaspase-9 and maintenance of the enzymatic activity of processed caspase-9. Furthermore, APAF1 is inactivated in metastatic melanomas, leading to defects in the execution of apoptotic cell death. Additionally, APAF1 has been shown to interact with NLRP1,5 Caspase-9, APIP, BCL2-like 1, and HSPA4.

 

Primary Antibody

Name: APAF1 IHC Antibody

Clone: Rabbit polyclonal

Supplier: IHC World

Catalog Number: IW-PA1249

Dilution: Ready to Use

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

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

Images: Search image

Additional Information:

Species Reactivity: Human, mouse, rat

Fixation: Formalin fixed paraffin embedded sections.

Positive Control: Rat brain

Negative Control: Omit primary antibody, isotype control, absorption control

Blocking: Normal serum blocking is not needed for this RTU antibody; 0.5-3% H2O2 to block endogenous peroxidase activity; avidin/biotin to block endogenous biotin activity if necessary

 

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