Description: HIF-1α Hypoxia contributes significantly to the pathophysiology of major categories of human disease, including myocardial and cerebral ischemia, cancer, pulmonary hypertension, congenital heart disease and chronic obstructive pulmonary disease. HIF-1 is a nuclear protein that activates gene transcription in response to reduced cellular O2 concentration. HIF-1 activates the transcription of EPO, VEGF, iNOS, heme oxygenase-1 and several other critical intracellular responses to hypoxia. HIF-1 is a heterodimer composed of HIF-1α and HIF-1β subunits. Both subunits are induced by hypoxia and rapidly decay upon return to normoxia. Recent research suggests that ability to regulate hypoxia-inducible factors may be related to tumor-related angiogenesis in certain cancers.
Primary Antibody
Name: HIF-1-alpha Antibody |
Clone: ESEE122, Mouse monoclonal |
Supplier: Novus Biologicals |
Catalog Number: NB100-131 |
Dilution: 1:1,000 - 1:10,000 depending on detection system 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 |
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: 45 minutes/20 minutes |
Standard Method: ABC Method or LSAB Method |
Enhanced Method: Polymeric Methods - Recommended methods for this HIF-1-alpha antibody |
Reagent: DAB |
Incubation Time/Temperature: 1-3 minutes/room temperature |
Reagent: Mayer's Hematoxylin |
Staining Time: 30 seconds |
Staining Pattern: Cytoplasmic in normoxia, nuclear translocation in response to hypoxia |
Images: Search image |
Species Reactivity: Mouse, rat, human, cow |
Fixation: Zinc fixed or Formalin fixed paraffin sections |
Positive Control: Mouse or human skin, hypoxia-induced tumors or human placenta |
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 |
1. Not tested on frozen sections. 2. Antigen retrieval with EDTA/Tris-EDTA produces high background staining and is NOT recommended for HIF-1 alpha staining. 3. When Clone ESEE122 used for mouse tissue, mouse on mouse blocking method is recommended to reduce unspecific background staining. 4. Standard ABC or LSAB method is NOT sensitive enough for this antibody so more sensitive detection systems are recommended such as ImmPRESS (Vector Labs) or Envision System (Dako). 5. Formalin fixed tissues tend to produce weaker staining so Zinc fixative is recommended for this antibody. |
References:
Zhong H, De Marzo AM, Laughner E, Lim M, Hilton DA, Zagzag D, Buechler P, Isaacs WB, Semenza GL, Simons JW. Overexpression of hypoxia-inducible factor 1alpha in common human cancers and their metastases. Cancer Res. 1999 Nov 15;59(22):5830-5.