Immunofluorescent analysis of HIF-1 alpha (green) in HeLa cells either left untreated (left panel) or treated with 100uM Deferoxamine mesylate for ~16 hours (right panel). Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 15 minutes at room temperature and blocked with 0.3% BSA for 15 minutes at room temperature. Cells were probed with a HIF-1 alpha monoclonal antibody (Product # MA1-516) at a dilution of 1:100 for at least 1 hour at room temperature, washed with PBS, and incubated with a DyLight 488-conjugated goat anti-mouse IgG secondary antibody (Product # 35502) at a dilution of 1:500 for 30 minutes at room temperature. F-actin (red) was stained with DyLight 594 Phalloidin (Product # 21836) and nuclei (blue) were stained with Hoechst 33342 dye (Product # 62249). Images were taken on a Thermo Scientific ArrayScan Instrument at 20X magnification.
|Tested species reactivity||Bovine, Human, Mouse, Non-human primate, Pig|
|Published species reactivity||Rabbit, Rat, Bovine, Mouse, Human|
|Host / Isotype||Mouse / IgG1|
|Immunogen||Human HIF-1 alpha amino acids 530-826.|
|Storage buffer||PBS, pH 7.4, with 1mg/ml BSA|
|Contains||0.05% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|Gel Shift (GS)||Assay dependent|
|Immunoelectrophoresis (IE)||Assay Dependent|
|Immunohistochemistry (Paraffin) (IHC (P))||1:10-1:100|
|Immunoprecipitation (IP)||Assay dependent|
|Western Blot (WB)||1:2000|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
MA1-516 detects hypoxia-inducible factor 1 alpha (HIF-1 alpha) from human, non-human primate, bovine, mouse and porcine cells. This antibody does not cross-react with ARNT or the related HIF-2 alpha.
MA1-516 has been successfully used in Western blot, immunofluorescence, immunoprecipitation, immunocytochemistry, and gel shift procedures. By Western blot, this antibody detects an ~116 kDa protein representing HIF-1 alpha after hypoxic induction in COS cells. Immunofluorescence staining of HIF-1 alpha in COS-7 cells with MA1-516 yields nuclear staining after exposing cells to 1% oxygen for 4 hours. In gel shift assays, MA1-516 has been successfully used only with mouse HIF-1 alpha.
Antibodies to this protein (and modification) were previously sold as part of a Thermo Scientific Cellomics High Content Screening Kit. This replacement antibody is now recommended for researchers who need an antibody for high content cell based assays. It has been thoroughly tested and validated for cellular immunofluorescence (IF) applications. Further optimization including the selection of the most appropriate fluorescent Dylight conjugated secondary antibody may have to be performed for your high content assay.
Oxygen availability plays a major role in the regulation of expression of many different genes including erythropoietin, nitric oxide synthase (NOS), heme oxygenase 1 (HO-1), glucose transporters and vascular growth factors necessary for the maintenance of homeostasis in hypoxic conditions. Hypoxia-inducible factor 1 alpha (HIF-1 alpha) has been identified as a bHLH-PAS family member which is instrumental in the oxygen-dependent regulation of these genes. HIF-1 alpha rapidly accumulates in the nucleus upon exposure to hypoxic conditions where it heterodimerizes with the aryl hydrocarbon nuclear receptor translocator, ARNT, also referred to as HIF-1 beta.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
A common functional regulatory variant at a type 2 diabetes locus upregulates ARAP1 expression in the pancreatic beta cell.
MA1-516 was used in EMSA to study the upregulation of pancreatic beta cell ARAP1 by a functional regulatory DNA variant at a type 2 diabetes locus
|Kulzer JR,Stitzel ML,Morken MA,Huyghe JR,Fuchsberger C,Kuusisto J,Laakso M,Boehnke M,Collins FS,Mohlke KL||American journal of human genetics (94:186)||2014|
Hypoxia-inducible factor and vascular endothelial growth factor are targets of dietary soy during acute stroke in female rats.
MA1-516 was used in western blot to study the role of HIF-1alpha and VEGF in the neuroprotective effects of soy following acute stroke
|Ma Y,Lovekamp-Swan T,Bekele W,Dohi A,Schreihofer DA||Endocrinology (154:1589)||2013|
Disrupting galectin-1 interactions with N-glycans suppresses hypoxia-driven angiogenesis and tumorigenesis in Kaposi's sarcoma.
MA1-516 was used in western blot to study the role of galectin-1 interactions with N-glycans in hypoxia-mediated angiogenesis and tumorigenesis in Kaposi's sarcoma
|Croci DO,Salatino M,Rubinstein N,Cerliani JP,Cavallin LE,Leung HJ,Ouyang J,Ilarregui JM,Toscano MA,Domaica CI,Croci MC,Shipp MA,Mesri EA,Albini A,Rabinovich GA||The Journal of experimental medicine (209:1985)||2012|
Up-regulation of glyceraldehyde-3-phosphate dehydrogenase gene expression by HIF-1 activity depending on Sp1 in hypoxic breast cancer cells.
MA1-516 was used in western blot to investigate the effect of HIF-1 on the regulation of GAPDH expression in hypoxic breast cancer cells
|Higashimura Y,Nakajima Y,Yamaji R,Harada N,Shibasaki F,Nakano Y,Inui H||Archives of biochemistry and biophysics (509:1)||2011|
Hypoxia preconditioning protection of corneal stromal cells requires HIF1alpha but not VEGF.
MA1-516 was used in western blot to investigate the role of HIF-1alpha on hypoxia preconditioning protection in corneal stromal cells
|Xing D,Bonanno JA||Molecular vision (15:1020)||2009|
Hypoxia upregulates carcinoembryonic antigen expression in cancer cells.
MA1-516 was used in western blot to study the upregulation of carcinoembryonic antigen expression in cancer cells by hypoxia
|Kokkonen N,Ulibarri IF,Kauppila A,Luosujärvi H,Rivinoja A,Pospiech H,Kellokumpu I,Kellokumpu S||International journal of cancer (121:2443)||2007|
Hypoxia preconditioning protects corneal stromal cells against induced apoptosis.
MA1-516 was used in western blot to investigate the influence of hypoxia on corneal stromal cell survival
|Xing D,Sun X,Li J,Cui M,Tan-Allen K,Bonanno JA||Experimental eye research (82:780)||2006|
Role of oxygen and vascular development in epithelial branching morphogenesis of the developing mouse lung.
MA1-516 was used in western blot to study the effect of oxygen level during mouse lung development
|van Tuyl M,Liu J,Wang J,Kuliszewski M,Tibboel D,Post M||American journal of physiology. Lung cellular and molecular physiology (288:L167)||2005|
Angiogenic potential of prostate carcinoma cells overexpressing bcl-2.
MA1-516 was used in western blot to investigate the role of bcl-2 in tumorigenesis
|Fernandez A,Udagawa T,Schwesinger C,Beecken W,Achilles-Gerte E,McDonnell T,D'Amato R||Journal of the National Cancer Institute (93:208)||2001|
Soluble FLT1 binds lipid microdomains in podocytes to control cell morphology and glomerular barrier function.
MA1-516 was used in blocking or activating experiment to study the role of the soluble isoform of FLT-1 in modulating podocyte/pericyte function
|Jin J,Sison K,Li C,Tian R,Wnuk M,Sung HK,Jeansson M,Zhang C,Tucholska M,Jones N,Kerjaschki D,Shibuya M,Fantus IG,Nagy A,Gerber HP,Ferrara N,Pawson T,Quaggin SE||Cell (151:384)||2012|
Experimental angiogenesis of arterial vasa vasorum.
MA1-516 was used in immunohistochemistry to study the development of arterial vasa vasorum in a rabbit model
|Bayer IM,Caniggia I,Adamson SL,Langille BL||Cell and tissue research (307:303)||2002|