Immunofluorescent analysis of HIF-1 alpha (green) in HeLa cells either left untreated (left panel) or treated with 0.1 mM Cobalt Chloride (right panel) for 4 hours. Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA (Product # 37525) for 15 minutes at room temperature. Cells were probed with a HIF-1 alpha polyclonal antibody (Product # PA1-16601) at a dilution of 1:100 for at least 1 hour at room temperature, washed with PBS, and incubated with DyLight 488 goat-anti-rabbit IgG secondary antibody (Product # 35552) at a dilution of 1:400 for 30 minutes at room temperature. F-Actin (red) was stained with DyLight-554 Phalloidin (Product # 21834) and nuclei (blue) were stained with Hoechst 33342 dye (Product # 62249). Images were taken on a Thermo Scientific ArrayScan at 20X magnification.
|Tested species reactivity||Goat, Hamster, Human, Mouse, Non-human primate, Rabbit, Rat|
|Published species reactivity||Rabbit, Rat, Human, Mouse, Not Applicable|
|Host / Isotype||Rabbit / IgG|
|Immunogen||A fusion protein including residues 530-825 of the mouse HIF-1 alpha protein.|
|Purification||Antigen affinity chromatography|
|Contains||0.05% sodium azide|
|Storage Conditions||4° C, do not freeze|
|Tested Applications||Dilution *|
|ChIP assay (ChIP)||Assay-Dependent|
|Flow Cytometry (Flow)||Assay-Dependent|
|Immunohistochemistry (IHC)||5-10 µg/ml|
|Immunohistochemistry (Frozen) (IHC (F))||5-10 µg/ml|
|Immunohistochemistry (Paraffin) (IHC (P))||5-10 µg/ml|
|Western Blot (WB)||1:500-1:2000|
* 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.
This antibody detects upregulation of HIF-1 alpha in hypoxic samples.
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 involved in mammalian oxygen homeostasis. This occurs as a posttranslational modification by prolyl hydroxylation. HIF-1 is a heterodimer composed of HIF-1 alpha and HIF-1 beta subunits. Both subunits are constantly translated. However, under normoxic conditions, human HIF-1 alpha is hydroxylated at Pro402 or Pro564 by a set of HIF prolyl hydroxylases, is polyubiquinated, and eventually degraded in proteosomes. Under hypoxic conditions, the lack of hydroxylation prevents HIF degradation and increases transcriptional activity. Therefore, the concentration of HIF-1 alpha increases in the cell. In contrast, HIF-1 beta remains stable under either condition. HIF hydroxylases provide insight into hypoxic cell responses, which may be used to help isolate therapeutic targets.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
|Not Applicable||Not Cited||
Metformin and resveratrol ameliorate muscle insulin resistance through preventing lipolysis and inflammation in hypoxic adipose tissue.
PA1-16601 was used in western blot to characterize prevention of lipolysis and inflammation in hypoxic adipose tissue by studying metformin and resveratrol that ameliorate muscle insulin resistance
|Zhao W,Li A,Feng X,Hou T,Liu K,Liu B,Zhang N||Cellular signalling (28:1401)||2016|
HIF-1¿-induced HSP70 regulates anabolic responses in articular chondrocytes under hypoxic conditions.
PA1-16601 was used in western blot to study the role of Hsp70 in HIF-1alpha-dependent anabolic pathways operating in hypoxic articular chondrocytes
|Tsuchida S,Arai Y,Takahashi KA,Kishida T,Terauchi R,Honjo K,Nakagawa S,Inoue H,Ikoma K,Ueshima K,Matsuki T,Mazda O,Kubo T||Journal of orthopaedic research : official publication of the Orthopaedic Research Society (32:975)||2014|
Inhibition of mitochondrial complex III blocks neuronal differentiation and maintains embryonic stem cell pluripotency.
PA1-16601 was used in western blot to study the suppression of the neuronal differentiation of murine embryonic stem cells following mitochondrial complex III inhibition
|Pereira SL,Grãos M,Rodrigues AS,Anjo SI,Carvalho RA,Oliveira PJ,Arenas E,Ramalho-Santos J||PloS one (8:null)||2013|
Response of small heat shock proteins in diabetic rat retina.
PA1-16601 was used in western blot to study the expression of small heat shock proteins in the retina of diabetic rats
|Reddy VS,Raghu G,Reddy SS,Pasupulati AK,Suryanarayana P,Reddy GB||Investigative ophthalmology and visual science (54:7674)||2013|
Superoxide induced by a high-glucose concentration attenuates production of angiogenic growth factors in hypoxic mouse mesenchymal stem cells.
PA1-16601 was used in western blot to investigate the interplay among high-glucose content, oxidative stress, and production of angiogenic growth factors in mesenchymal stem cells
|Ishizuka T,Hinata T,Watanabe Y||The Journal of endocrinology (208:147)||2011|
The Neuroprotective Effect of Coumaric Acid on Spinal Cord Ischemia/Reperfusion Injury in Rats.
PA1-16601 was used in immunohistochemistry - paraffin section to demonstrate that coumaric acid is protective after a ischemia-reperfusion injury of the spinal cord.
|Guven M,Sehitoglu MH,Yuksel Y,Tokmak M,Aras AB,Akman T,Golge UH,Karavelioglu E,Bal E,Cosar M||Inflammation (38:1986)||2015|
Wnt signaling directs a metabolic program of glycolysis and angiogenesis in colon cancer.
PA1-16601 was used in immunohistochemistry to study the role of PDK1 in the mechanism by which Wnt signaling modulates glycolytic flux and angiogenesis in colon cancer
|Pate KT,Stringari C,Sprowl-Tanio S,Wang K,TeSlaa T,Hoverter NP,McQuade MM,Garner C,Digman MA,Teitell MA,Edwards RA,Gratton E,Waterman ML||The EMBO journal (33:1454)||2014|