使用敲除细胞株进行验证
重组
RabMAb

Anti-Hsp27抗体[EPR5477] (ab109376)

概述

  • 产品名称
    Anti-Hsp27抗体[EPR5477]
    参阅全部 Hsp27 一抗
  • 描述
    兔单克隆抗体[EPR5477] to Hsp27
  • 经测试应用
    适用于: ICC/IF, WB, IP, IHC-P, Flow Cytmore details
  • 种属反应性
    与反应: Mouse, Rat, Human
  • 免疫原

    A synthetic peptide corresponding to residues in Human Hsp27

  • 阳性对照
    • HeLa, HAP1, COS-1, BxPC-3 and HT-1376 cell lysates
  • 常规说明

     

     

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents

    This product is a recombinant rabbit monoclonal antibody.

性能

应用

Our Abpromise guarantee covers the use of ab109376 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

应用 Ab评论 说明
ICC/IF 1/500.
WB 1/1000 - 1/10000. Predicted molecular weight: 23 kDa.Can be blocked with Hsp27 peptide (ab215449).
IP 1/10 - 1/100.
IHC-P 1/100 - 1/250. Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol.
Flow Cyt 1/10 - 1/100.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

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.
  • Information by UniProt
  • 数据库链接
  • 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



  • Predicted band size : 23 kDa

    Lane 1: Wild type HAP1 whole cell lysate (20 µg)
    Lane 2: Hsp27 knockout  HAP1 whole cell lysate (20 µg)
    Lane 3: HeLa whole cell lysate (20 µg)
    Lane 4: MCF7 whole cell lysate (20 µg)

    Lanes 1 - 4: Merged signal (red and green). Green - ab109376 observed at 27 kDa. Red - loading control, ab8245, observed at 37 kDa.

    ab109376 was shown to specifically react with Hsp27 in wild-type cells as signal was lost in Hsp27 knockout HEP1 cells. Wild-type and Hsp27 knockout samples were subjected to SDS-PAGE.  Ab109376 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

  • Immunocytochemistry/Immunofluorescence analysis of HeLa (human cervix adenocarcinoma) cells labelling Hsp27 with purified ab109376 at 1/500. Cells were fixed with 100% methanol. ab150077, Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. Nuclei were counterstained with DAPI (blue).

    Secondary Only Control: PBS was used instead of the primary antibody as the negative control.

  • All lanes : Anti-Hsp27 antibody [EPR5477] (ab109376) at 1/1000 dilution

    Lane 1 : HeLa cell lysate
    Lane 2 : COS-1 cell lysate
    Lane 3 : BxPC-3 cell lysate
    Lane 4 : HT-1376 cell lysate

    Lysates/proteins at 10 µg per lane.


    Predicted band size : 23 kDa
    Observed band size : 27 kDa (why is the actual band size different from the predicted?)

文献

ab109376 has not yet been referenced specifically in any publications.

客户评价及客户问答

Application
Western blot
Sample
Human Cell lysate - whole cell (BEL-7402)
Gel Running Conditions
Non-reduced Denaturing (10%)
Loading amount
50 µg
Treatment
80 µM Doxorubicin for 24hrs
Specification
BEL-7402
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Dr. guoqing zhu

Verified customer

提交于 Jan 26 2016

Application
Western blot
Loading amount
62.5 µg
Gel Running Conditions
Reduced Denaturing (15% acrylamide)
Sample
Pig Tissue lysate - other (Muscle sarcoplasmic extract)
Specification
Muscle sarcoplasmic extract
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Username

Dr. Shannon Cruzen

Verified customer

提交于 Feb 14 2014

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

注册