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HIF-1alpha (D1S7W) XP® Rabbit mAb
HIF-1alpha (D1S7W) XP® Rabbit mAb

HIF-1alpha (D1S7W) XP® Rabbit mAb

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    分子量：
    120
    120
    反应种属：
    Human,Mouse,Monkey,
    Human,Mouse,Monkey,
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    产品介绍
    产品信息
    抗原名称
    HIF-1alpha
    来源纯化
    使用与人源 HIF-1α 蛋白的 Lys478 周围的残基相对应的合成肽对动物进行免疫接种来产生单克隆抗体。
    宿主
    Rabbit
    简单描述
    Highly specific and rigorously validated rabbit monoclonal anti HIF-1 alpha antibody (D1S7W). Validated for WB, IP, IF, F, ChIP, ChIP, C&R. Click here.
    商品描述

    Product Usage Information

    For optimal ChIP and ChIP-seq results, use 5 μl of antibody and 10 μg of chromatin (approximately 4 x 106 cells) per IP. This antibody has been validated using SimpleChIP® Enzymatic Chromatin IP Kits. The CUT&RUN dilution was determined using CUT&RUN Assay Kit #86652.

    ApplicationDilution
    Western Blotting1:1000
    Immunoprecipitation1:50
    Immunofluorescence (Immunocytochemistry)1:400 - 1:1600
    Flow Cytometry (Fixed/Permeabilized)1:100 - 1:400
    Chromatin IP1:100
    Chromatin IP-seq1:100
    CUT&RUN1:100
    分子量
    120
    研究领域
    癌症,细胞生物学,纤维化,代谢,神经科学
    应用
    反应种属
    Human,Mouse,Monkey,
    目标/特异性

    Specificity/Sensitivity

    HIF-1α (D1S7W) XP® Rabbit mAb 可识别内源水平的 HIF-1α 总蛋白。这种抗体不与 HIF-2α 蛋白发生交叉反应。

    物种反应性：

    人, 小鼠, 猴

    敏感性
    Endogenous
    背景
    背景
    低氧诱导因子 1 (HIF1) 是在细胞对缺氧的反应中发挥至关重要作用的异二聚转录因子 (1)。HIF1 复合体由作为 PAS（Per、ARNT、Sim）家族碱性螺旋-环-螺旋蛋白的两个亚基 HIF-1α 和 HIF-1β 组成 (2)。HIF1 调节诸多基因类型转录，这些基因可促进低氧环境的反应，包括调节血管生成、红细胞生成、细胞周期、代谢和凋亡的基因。广泛表达的 HIF-1α 一般在常氧细胞中可通过泛素/蛋白酶体通路快速降解。在常氧条件下，HIF-1α 会发生脯氨酸羟基化，这会导致能够促进与希佩尔-林道 (VHL) 蛋白 E3 连接酶复合体结合的构象变化；随后发生泛素化和蛋白酶体降解 (3,4)。低氧条件和羟化酶化学抑制剂（如去铁敏和钴）均可抑制 HIF-1α 降解并且导致其稳定。此外，HIF-1α 可按不依赖于氧的方式，由各种细胞因子，经 PI3K-AKT-mTOR 通路诱导 (5-7)。HIF-1β 也称作 AhR 胞核易位蛋白 (ARNT)，原因在于其具有与芳基烃受体 (AhR) 匹配以形成异二聚转录因子复合体的能力 (8)。HIF-1β 与 AhR 均可在生物异源物质代谢中发挥重要作用 (8)。此外，导致 TEL-ARNT 融合蛋白的染色体易位与急性成髓细胞性白血病相关 (9)。研究还表明，ARNT/HIF-1β 表达水平在2 型糖尿病患者的胰岛中显著下降，这表明 HIF-1β 在胰腺 β-细胞功能中发挥重要作用 (10)。 1.Sharp, F.R. and Bernaudin, M. (2004) Nat Rev Neurosci 5, 437-48. 2.Wang, G.L. et al. (1995) Proc Natl Acad Sci U S A 92, 5510-4. 3.Jaakkola, P. et al. (2001) Science 292, 468-72. 4.Maxwell, P.H. et al. (1999) Nature 399, 271-5. 5.Fukuda, R. et al. (2002) J Biol Chem 277, 38205-11. 6.Jiang, B.H. et al. (2001) Cell Growth Differ 12, 363-9. 7.Laughner, E. et al. (2001) Mol Cell Biol 21, 3995-4004. 8.Walisser, J.A. et al. (2004) Proc Natl Acad Sci U S A 101, 16677-82. 9.Salomon-Nguyen, F. et al. (2000) Proc Natl Acad Sci U S A 97, 6757-62. 10.Gunton, J.E. et al. (2005) Cell 122, 337-49.
    研究领域
    癌症,细胞生物学,纤维化,代谢,神经科学
    翻译后修饰
    Unmodified
    制备和贮存
    保存方式
    Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.For a carrier free (BSA and azide free) version of this product see product #36199.
    数据库链接
    Entrez-Gene ID
    3091
    UniProt ID
    Q16665

    参考图片

    Western blot analysis of extracts from Hep G2 cells untreated (-) or treated with cobalt chloride (100 µM, 4 h; +), Raji cells untreated (-) or treated with cobalt chloride (100 µM, 4 h; +) and U-2 OS cells untreated (-) or treated with DMOG (1 mM, 6 h; +) using HIF-1α (D1S7W) XP® Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).

    Immunoprecipitation of HIF-1α from lysate of Hep G2 cells treated with cobalt chloride (100 µM, 4 h). Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is HIF-1α (D1S7W) XP® Rabbit mAb. Western blot analysis was performed using HIF-1α (D1S7W) XP® Rabbit mAb. Anti-rabbit IgG, HRP-linked Antibody #7074 was used as the secondary antibody.

    Confocal immunofluorescent analysis of Hep G2 cells, untreated (left) or treated with cobalt chloride (500 μM, 24 h; right), using HIF-1α (D1S7W) XP® Rabbit mAb (green). Actin filaments were labeled with DyLight™ 554 Phalloidin #13054 (red).

    Flow cytometric analysis of U-2 OS cells, untreated (blue) or treated with DMOG (1 mM, 6 h; green), using HIF-1α (D1S7W) XP® Rabbit mAb (solid lines) or concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype control #3900 (dashed lines). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.

    Chromatin immunoprecipitations were performed with cross-linked chromatin from MCF7 cells treated with cobalt chloride (100 μM) overnight and HIF-1α (D1S7W) XP® Rabbit mAb, using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. DNA Libraries were prepared from 5 ng enriched ChIP DNA using NEBNext® Ultra™ II DNA Library Prep Kit for Illumina®, and sequenced on the Illumina NextSeq. The figure shows binding across ARRDC3, a known target gene of HIF-1α (see additional figure containing ChIP-qPCR data). For additional ChIP-seq tracks, please download the product data sheet.

    Chromatin immunoprecipitations were performed with cross-linked chromatin from MCF7 cells treated with cobalt chloride (100 μM, overnight) and either HIF-1α (D1S7W) XP® Rabbit mAb or Normal Rabbit IgG #2729, using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. The enriched DNA was quantified by real-time PCR using human ARRDC3 downstream primers, SimpleChIP® Human ERRFI1 Upstream Primers #31180, and SimpleChIP® Human α Satellite Repeat Primers #4486. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.

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    货号：
    36169T
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