IGF-IR Antibody Staining Protocol for Immunohistochemistry

 

 

 

Description: Receptor tyrosine kinases (RTKs) are transmembrane molecular scaffolds that influence cellular processes including the cell cycle, cell migration, cell metabolism survival, proliferation and differentiation. Insulin-like growth factor-I receptor (IGF-IR) is an RTK that stimulates growth in many different cell types, blocks apoptosis, acts as an intermediate of many growth hormone responses, and may stimulate the growth of some types of cancer. The IGF-IR cognate ligand insulin-like growth factor I (IGF-1) promotes association of IGF-IR with Shc, Grb2, and Sos-1, which initiates Ras and Erk kinase cascades, thereby modifying transcription factor activity, such as activation of the ELK transcription factors. The modular phosphotyrosine binding (PTB) domains of insulin receptor substrate (IRS) -1 and -2 can associate with active IGF1-R and initiate phosphatidylinositol 3-kinase-dependent downstream signals. The human IGF-IR gene maps to chromosome 15q25-q26 and encodes a 1376 amino acid precursor protein that cleaves into alpha and beta subunits. The human IGF-IIR gene maps to chromosome 6q26 and encodes a 2491 amino acid, 250 kDa transmembrane protein.

 

Primary Antibody

Name: IGF-IR Antibody

Clone: Rabbit anti-Human

Supplier: Santa Cruz Biotechnology

Catalog Number: sc-9038

Dilution: 1:200 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 minutes/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 ºC - 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: Nuclear

Images: Search image

Additional Information:

Tissue Type: Prostate

Fixation: Formalin fixed paraffin sections

Positive Control: Prostate

Negative Control: Omit primary antibody, isotype control or 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

Notes:

Not tested on frozen sections.

 

References: