HIF-2 alpha Rabbit Recombinant mAb

目录号：A5100

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规格	价格	库存
20ul	RMB 447.68	现货
100ul	RMB 1500.41	现货
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使用信息

抗体应用

IF,ELISA

稀释比例

1:50

反应性

Human Mouse Rat

MW (kDa)

96kDa

抗体类型

Rabbit

浓度

1mg/ml

储存液配方

10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide.

储存条件
(自收到货起)

Store at –20°C.

Datasheet & SDS

生物描述

特异性	HIF-2 alpha Rabbit Recombinant mAb detects endogenous levels of total HIF-2.
背景	Hypoxia is defined as a reduction of oxygen amount available to a cell, tissue or organism. Hypoxia inducible factors (HIFs), are the master regulators of oxygen homeostasis and play a role in development, postnatal physiology as well as disease pathogenesis. Two known hypoxia-inducible factors (HIFs), HIF-1α and HIF-2α, primarily mediate the transcriptional response to hypoxia. Despite the high homology between HIF-1α and HIF-2α, emerging evidence suggests differences between both molecules in terms of transcriptional targets as well as impact on multiple physiological pathways and tumorigenesis. HIF-2 is a heterologous dimeric protein complex that consists of two subunits: HIF-2α and HIF-1β. Both HIF-2α and HIF-1β belong to the superfamily of transcription factors bHLH-PAS. HIF-2α expression is restricted to specific types, including endothelial cells, glial cells, type II pneumocytes, cardiomyocytes, kidney fibroblasts, interstitial cells of the pancreas and duodenum and hepatocytes. It is confined to the cell nucleus and expressed only under hypoxic stimulation. HIF-2α and HIF-1α regulate key downstream genes, including c-Myc, p53, mTOR and β-catenin, conversely. HIF-2α induction promotes cell cycle progression by enhancing c-Myc function. HIF-2α plays a critical role in metabolism, modulating the expression of cytochrome c oxidase isoforms so as to maximize the efficiency of the electron transport chain. Defects in this response lead to impaired ATP production and elevated oxidant production in hypoxia.

特异性

HIF-2 alpha Rabbit Recombinant mAb detects endogenous levels of total HIF-2.

背景

Hypoxia is defined as a reduction of oxygen amount available to a cell, tissue or organism. Hypoxia inducible factors (HIFs), are the master regulators of oxygen homeostasis and play a role in development, postnatal physiology as well as disease pathogenesis. Two known hypoxia-inducible factors (HIFs), HIF-1α and HIF-2α, primarily mediate the transcriptional response to hypoxia. Despite the high homology between HIF-1α and HIF-2α, emerging evidence suggests differences between both molecules in terms of transcriptional targets as well as impact on multiple physiological pathways and tumorigenesis. HIF-2 is a heterologous dimeric protein complex that consists of two subunits: HIF-2α and HIF-1β. Both HIF-2α and HIF-1β belong to the superfamily of transcription factors bHLH-PAS. HIF-2α expression is restricted to specific types, including endothelial cells, glial cells, type II pneumocytes, cardiomyocytes, kidney fibroblasts, interstitial cells of the pancreas and duodenum and hepatocytes. It is confined to the cell nucleus and expressed only under hypoxic stimulation. HIF-2α and HIF-1α regulate key downstream genes, including c-Myc, p53, mTOR and β-catenin, conversely. HIF-2α induction promotes cell cycle progression by enhancing c-Myc function. HIF-2α plays a critical role in metabolism, modulating the expression of cytochrome c oxidase isoforms so as to maximize the efficiency of the electron transport chain. Defects in this response lead to impaired ATP production and elevated oxidant production in hypoxia.

实验方法

Immunofluorescence (Immunocytochemistry)

Specimen Preparation (forcultured cell lines, IF-IC)

1. Aspirate liquid, then cover cells to a depth of 2–3 mm with 4% formaldehyde diluted in 1X PBS.
NOTE: Formaldehyde is toxic, use only in a fume hood.
2. Fix cells for 15 min at room temperature.
3. Aspirate fixative, rinse three times in 1X PBS for 5 min each.
4. Proceed with Immunostaining.

Immunostaining

1. Add theblocking buffer and incubate for 60 min at RT.
2. Prepare primary antibody diluent in antibody dilution buffer as recommended .
3. Aspirate blocking solution, apply diluted primary antibody.
4. Incubate overnight at 4°C.
5. Rinse three times in 1X PBS for 5 min each.
6. Incubate specimens in fluorochrome-conjugated secondary antibody diluted in antibody dilution buffer for 1–2 hr at room temperature in the dark.
7. Rinse three times in 1X PBS for 5 min each.
8. Mount slides usingmounting medium with DAPI and cover with coverslips.
9. For best results, allow mountant to cure overnight at room temperature. For long-term storage, store slides flat at 4°C protected from light.

参考文献	[1] Zhao J, et al. Cell Death Dis. 2015, 6:e1600. [2] Loboda A, et al. Mol Cells. 2010, 29(5):435-42. [3] Bertout JA, et al. Proc Natl Acad Sci U S A. 2009, 106(34):14391-6.

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