APPLICATION
Application Note
HIF1a (Hypoxia-inducible factor 1-alpha) is a subunit of a transcription factor, which functions as a master transcriptional regulator of the adaptive response to hypoxia. The protein chaperone heat shock protein 90 (Hsp90) is a major regulator of different transcription factors, including HIF1a, thus a binding ELISA assay was conducted to detect the interaction of HIF1a and HSP90. Briefly, recombinant human HIF1a were diluted serially in PBS, with 0.01% BSA (pH 7.4). Duplicate samples of 100 μl were then transferred to HSP90-coated microtiter wells and incubated for 2h at 37ºC. Wells were washed with PBST and incubated for 1h with anti-HIF1a pAb, then aspirated and washed 3 times. After incubation with HRP labelled secondary antibody, wells were aspirated and washed 3 times. With the addition of substrate solution, wells were incubated 15-25 minutes at 37ºC. Finally, add 50 μl stop solution to the wells and read at 450nm immediately. The binding activity of of HIF1a and HSP90 was in a dose dependent manner.
Observed MW
40 kDa.
PROPERTIES
Form
Lyophilized powder
Buffer
Reconstitute with 20mM Tris and 150mM NaCl to 0.1-1.0mg/ml. Do not vortex. Lyophilized from 20mM Tris, 150mM NaCl, 1mM EDTA, 1mM DTT, 0.01% SKL, 5% Trehalose.
Preservative
ProClin 300
Storage
For short-term storage (1-2 weeks), store at 4ºC. For long-term storage, store at -20ºC or below. After reconstitution, keep as concentrated solution. Avoid freeze-thaw cycles.
Region/Sequence
N-terminal His-Tag; Thr218~Thr506 (NP_001230013.1)
Expression System
E. coli
Purity
> 95%
Endotoxin
< 1 EU/μg
Conjugation
Unconjugated
Note
For laboratory use only. Not for any clinical, therapeutic, or diagnostic use in humans or animals. Not for animal or human consumption.
TARGET
Synonyms
hypoxia inducible factor 1 subunit alpha , HIF-1-alpha , HIF-1A , HIF-1alpha , HIF1 , HIF1-ALPHA , MOP1 , PASD8 , bHLHe78
Background
This gene encodes the alpha subunit of transcription factor hypoxia-inducible factor-1 (HIF-1), which is a heterodimer composed of an alpha and a beta subunit. HIF-1 functions as a master regulator of cellular and systemic homeostatic response to hypoxia by activating transcription of many genes, including those involved in energy metabolism, angiogenesis, apoptosis, and other genes whose protein products increase oxygen delivery or facilitate metabolic adaptation to hypoxia. HIF-1 thus plays an essential role in embryonic vascularization, tumor angiogenesis and pathophysiology of ischemic disease. Alternatively spliced transcript variants encoding different isoforms have been identified for this gene. [provided by RefSeq, Jul 2011]
Database
Research Area
DATA IMAGES
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GTX00119-pro Functional Assay Image
Functional ELISA analysis of GTX00119-pro Human HIF1 alpha protein which can bind immobilized HSP90 protein.
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GTX00119-pro Image
SDS-PAGE analysis of GTX00119-pro Human HIF1 alpha protein.
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GTX00119-pro Image
WB analysis of GTX00119-pro Human HIF1 alpha protein.
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REFERENCE
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REVIEW
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