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Signal Transduction Protein Trafficking Chaperones Heat Shock Proteins

Anti-Hsp27 (phospho S85)抗体(ab5594)

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Hsp27 (phospho S85) antibody (ab5594)
  • Immunocytochemistry - Anti-Hsp27 (phospho S85) antibody (ab5594)
  • Western blot - Anti-Hsp27 (phospho S85) antibody (ab5594)
  • Immunoprecipitation - Anti-Hsp27 (phospho S85) antibody (ab5594)
  • Western blot - Anti-Hsp27 (phospho S85) antibody (ab5594)

Key features and details

  • Rabbit polyclonal to Hsp27 (phospho S85)
  • Suitable for: ICC, IHC-P, IP, WB
  • Reacts with: Rat, Human
  • Isotype: IgG

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概述

  • 产品名称

    Anti-Hsp27 (phospho S85)抗体
    参阅全部 Hsp27 一抗
  • 描述

    兔多克隆抗体to Hsp27 (phospho S85)
  • 宿主

    Rabbit
  • 经测试应用

    适用于: ICC, IHC-P, IP, WBmore details
  • 种属反应性

    与反应: Rat, Human
    预测可用于: Mouse, Cow, Dog, Pig, Xenopus laevis, Chinese hamster
  • 免疫原

    Synthetic peptide corresponding to Rat Hsp27 aa 50-150 (phospho S85).

    Run BLAST with BLAST the sequence with ExPASy Run BLAST with BLAST the sequence with NCBI
  • 常规说明

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.

    If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

性能

  • 形式

    Liquid
  • 存放说明

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
  • 存储溶液

    Preservative: 0.05% Sodium azide
    Constituents: PBS, 3% BSA
  • Concentration information loading...
  • 纯度

    Immunogen affinity purified
  • 克隆

    多克隆
  • 同种型

    IgG
  • 研究领域

    • Signal Transduction
    • Protein Trafficking
    • Chaperones
    • Heat Shock Proteins
    • Cancer
    • Tumor biomarkers
    • Other
    • Cardiovascular
    • Atherosclerosis
    • Ischemia / Reperfusion
    • Cardiovascular
    • Heart
    • Contractility
    • Contractile Proteins
    • Actins

相关产品

  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
  • Isotype control

    • Rabbit IgG, polyclonal - Isotype Control (ChIP Grade) (ab171870)
  • Recombinant Protein

    • Recombinant Human Hsp27 (phospho S82) protein (ab113185)

应用

The Abpromise guarantee

Abpromise™承诺保证使用ab5594于以下的经测试应用

“应用说明”部分 下显示的仅为推荐的起始稀释度;实际最佳的稀释度/浓度应由使用者检定。

应用 Ab评论 说明
ICC
Use a concentration of 15 - 25 µg/ml.
IHC-P
1/100. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
IP
Use at 6 µg/mg of lysate.
WB (1)
Use a concentration of 0.25 - 2 µg/ml. Detects a band of approximately 27 kDa.
说明
ICC
Use a concentration of 15 - 25 µg/ml.
IHC-P
1/100. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
IP
Use at 6 µg/mg of lysate.
WB
Use a concentration of 0.25 - 2 µg/ml. Detects a band of approximately 27 kDa.

靶标

  • 功能

    Involved in stress resistance and actin organization.
  • 组织特异性

    Detected in all tissues tested: skeletal muscle, heart, aorta, large intestine, small intestine, stomach, esophagus, bladder, adrenal gland, thyroid, pancreas, testis, adipose tissue, kidney, liver, spleen, cerebral cortex, blood serum and cerebrospinal fluid. Highest levels are found in the heart and in tissues composed of striated and smooth muscle.
  • 疾病相关

    Defects in HSPB1 are the cause of Charcot-Marie-Tooth disease type 2F (CMT2F) [MIM:606595]. CMT2F is a form of Charcot-Marie-Tooth disease, the most common inherited disorder of the peripheral nervous system. Charcot-Marie-Tooth disease is classified in two main groups on the basis of electrophysiologic properties and histopathology: primary peripheral demyelinating neuropathy or CMT1, and primary peripheral axonal neuropathy or CMT2. Neuropathies of the CMT2 group are characterized by signs of axonal regeneration in the absence of obvious myelin alterations, normal or slightly reduced nerve conduction velocities, and progressive distal muscle weakness and atrophy. Nerve conduction velocities are normal or slightly reduced. CMT2F onset is between 15 and 25 years with muscle weakness and atrophy usually beginning in feet and legs (peroneal distribution). Upper limb involvement occurs later. CMT2F inheritance is autosomal dominant.
    Defects in HSPB1 are a cause of distal hereditary motor neuronopathy type 2B (HMN2B) [MIM:608634]. Distal hereditary motor neuronopathies constitute a heterogeneous group of neuromuscular disorders caused by selective impairment of motor neurons in the anterior horn of the spinal cord, without sensory deficit in the posterior horn. The overall clinical picture consists of a classical distal muscular atrophy syndrome in the legs without clinical sensory loss. The disease starts with weakness and wasting of distal muscles of the anterior tibial and peroneal compartments of the legs. Later on, weakness and atrophy may expand to the proximal muscles of the lower limbs and/or to the distal upper limbs.
  • 序列相似性

    Belongs to the small heat shock protein (HSP20) family.
  • 翻译后修饰

    Phosphorylated in MCF-7 cells on exposure to protein kinase C activators and heat shock.
  • 细胞定位

    Cytoplasm. Nucleus. Cytoplasm > cytoskeleton > spindle. Cytoplasmic in interphase cells. Colocalizes with mitotic spindles in mitotic cells. Translocates to the nucleus during heat shock and resides in sub-nuclear structures known as SC35 speckles or nuclear splicing speckles.
  • Target information above from: UniProt accession P04792 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • 数据库链接

    • Entrez Gene: 516099 Cow
    • Entrez Gene: 3315 Human
    • Entrez Gene: 15507 Mouse
    • Entrez Gene: 493184 Pig
    • Entrez Gene: 24471 Rat
    • Omim: 602195 Human
    • SwissProt: Q3T149 Cow
    • SwissProt: P04792 Human
    • SwissProt: P14602 Mouse
    • SwissProt: Q5S1U1 Pig
    • SwissProt: P42930 Rat
    • Unigene: 520973 Human
    • Unigene: 13849 Mouse
    • Unigene: 3841 Rat
    see all
  • 别名

    • Heat shock 27kDa protein antibody
    • 28 kDa heat shock protein antibody
    • CMT2F antibody
    • DKFZp586P1322 antibody
    • epididymis secretory protein Li 102 antibody
    • Estrogen regulated 24 kDa protein antibody
    • Estrogen-regulated 24 kDa protein antibody
    • Heat shock 25kDa protein 1 antibody
    • Heat shock 27 kDa protein antibody
    • Heat shock 27kD protein 1 antibody
    • Heat shock 27kDa protein 1 antibody
    • Heat shock 28kDa protein 1 antibody
    • Heat Shock Protein 27 antibody
    • Heat shock protein beta 1 antibody
    • Heat shock protein beta-1 antibody
    • heat shock protein family B (small) member 1 antibody
    • HEL-S-102 antibody
    • HMN2B antibody
    • HS.76067 antibody
    • Hsp 25 antibody
    • HSP 27 antibody
    • Hsp 28 antibody
    • Hsp B1 antibody
    • Hsp25 antibody
    • HSP27 antibody
    • Hsp28 antibody
    • HspB1 antibody
    • HSPB1_HUMAN antibody
    • SRP27 antibody
    • Stress responsive protein 27 antibody
    • Stress-responsive protein 27 antibody
    see all

图片

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Hsp27 (phospho S85) antibody (ab5594)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Hsp27 (phospho S85) antibody (ab5594)

    Immunohistochemistry analysis of paraffin-embedded human skeletal muscle tissue labelling Hsp27 (phospho S85) with ab5594 (right) compared to a negative control without primary antibody (left).

    To expose target proteins, antigen retrieval was performed using 10mM sodium citrate (pH 6.0), microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS, and then probed with ab5594 diluted in 3% BSA-PBS at 1/100 dilution for 1 hour at 37°C in a humidified chamber. Tissues were washed extensively in PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.

  • Immunocytochemistry - Anti-Hsp27 (phospho S85) antibody (ab5594)
    Immunocytochemistry - Anti-Hsp27 (phospho S85) antibody (ab5594)

    Immunocytochemistry/Immunofluorescence analysis of Hsp27 (phospho S85) (green) in HeLa cells either left untreated (left panel) or treated with 10uM Anisomysin (right panel) for 30 minutes. 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 for 15 minutes at room temperature. Cells were probed with ab5594 at 20ug/ml for at least 1 hour at room temperature, washed with PBS, and incubated with DyLight 488 goat anti-rabbit IgG secondary antibody at 1:400 for 30 minutes at room temperature. F-Actin (red) was stained with DyLight 554 Phalloidin and nuclei (blue) were stained with Hoechst 33342 dye. 20X magnification.

  • Western blot - Anti-Hsp27 (phospho S85) antibody (ab5594)
    Western blot - Anti-Hsp27 (phospho S85) antibody (ab5594)
    All lanes : Anti-Hsp27 (phospho S85) antibody (ab5594) at 2 µg/ml

    Lane 1 : Hela cell lysate - untreated
    Lane 2 : Hela cell lysate - treated with 10uM Anisomycin for 30 minutes

    Lysates/proteins at 50 µg per lane.

    Secondary
    All lanes : HRP-conjugated goat anti-rabbit IgG at 1/20000 dilution
  • Immunoprecipitation - Anti-Hsp27 (phospho S85) antibody (ab5594)
    Immunoprecipitation - Anti-Hsp27 (phospho S85) antibody (ab5594)

    Immunoprecipitation of Hsp27 (phospho S85) was performed on HeLa cells treated with 10uM Anisomysin for 30 minutes. Antigen-antibody complexes were formed by incubating 500µg of whole cell lysate with 3µg of ab5594 overnight on a rocking platform at 4°C. The immune complexes were captured on 50µl Protein A/G Agarose, washed extensively, and eluted with Lane Marker Reducing Sample Buffer. HeLa cell lysate (50ug) was loaded as a positive control (left lane). Samples were resolved on a 4-20% Tris-HCl polyacrylamide gel, transferred to a PVDF membrane, and blocked with 5% BSA/TBST for at least 1 hour. The membrane was probed with ab5594 at 2ug/ml overnight rotating at 4°C, washed in TBST, and probed with Clean-Blot IP Detection Reagent at a dilution of 1:1000 for at least 1 hour. Chemiluminescent detection was performed using SuperSignal West Dura.

  • Western blot - Anti-Hsp27 (phospho S85) antibody (ab5594)
    Western blot - Anti-Hsp27 (phospho S85) antibody (ab5594)
    Western blot analysis of Hsp27 (phospho S85) was performed by loading rat skeletal muscle tissue extracts on a SDS-PAGE gel. Proteins were transferred to a membrane probed with a Hsp27 (phospho S85) polyclonal antibody (ab5594) at a concentration of 0.25ug/ml followed by an anti-rabbit IgG-HRP secondary antibody. Detection was performed using a chemiluminescent substrate.

实验方案

  • Immunoprecipitation protocols
  • Immunocytochemistry & immunofluorescence protocols
  • Western blot protocols

Click here to view the general protocols

数据表及文件

  • Datasheet download

    Download

文献 (11)

发表研究结果有使用 ab5594?请让我们知道,以便我们可以引用本数据表中的参考文章。

ab5594 被引用在 11 文献中.

  • Matsuo K  et al. Geranylgeranylacetone attenuates cerebral ischemia-reperfusion injury in rats through the augmentation of HSP 27 phosphorylation: a preliminary study. BMC Neurosci 22:9 (2021). PubMed: 33557752
  • Yao K  et al. HMGN5 promotes IL-6-induced epithelial-mesenchymal transition of bladder cancer by interacting with Hsp27. Aging (Albany NY) 12:7282-7298 (2020). PubMed: 32315283
  • Wenzina J  et al. Inhibition of p38/MK2 Signaling Prevents Vascular Invasion of Melanoma. J Invest Dermatol 140:878-890.e5 (2020). PubMed: 31622599
  • Yamamoto Y  et al. Pentose phosphate pathway activation via HSP27 phosphorylation by ATM kinase: A putative endogenous antioxidant defense mechanism during cerebral ischemia-reperfusion. Brain Res 1687:82-94 (2018). WB ; Rat . PubMed: 29510140
  • Zhai W  et al. A1 adenosine receptor attenuates intracerebral hemorrhage-induced secondary brain injury in rats by activating the P38-MAPKAP2-Hsp27 pathway. Mol Brain 9:66 (2016). Rat . PubMed: 27301321
View all Publications for this product

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Western blot abreview for Anti-Hsp27 (phospho S85) antibody

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Rat Cell lysate - whole cell (primary hepatocyte)
Specification
primary hepatocyte
Blocking step
Other as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: ?
Read More

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