概述

  • 产品名称Anti-Hsp27抗体[G3.1]
    参阅全部 Hsp27 一抗
  • 描述
    小鼠单克隆抗体[G3.1] to Hsp27
  • 经测试应用适用于: Flow Cyt, IHC-Fr, ICC/IF, IHC-P, WBmore details
  • 种属反应性
    与反应: Mouse, Rat, Sheep, Chicken, Human, Non human primates
    预测可用于: Cow, Pig
  • 免疫原

    Full length native protein (purified) corresponding to Human Hsp27. (Partially purified human HSP27)

  • 阳性对照
    • Hela Cells.

性能

应用

Our Abpromise guarantee covers the use of ab2790 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
Flow Cyt 1/100. ab170190-Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
IHC-Fr 1/100.
ICC/IF Use a concentration of 0.5 - 1 µg/ml.
IHC-P Use a concentration of 0.5 - 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

Use citrate buffer.

WB 1/1000.

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

Anti-Hsp27 antibody [G3.1] 图像

  • Immunocytochemistry/ Immunofluorescence analysis of non small cell lung carcinoma NCI-H1299 cells labeling Hsp27 with ab2790 at 1/300 dilution. The cells were fixed with formaldehyde and permeabilised with 0.2% TritonX-100. The cells were blocked with 1.5% serum for 10 minutes at 25°C, followed by incubation with Anti-Hsp27 antibody [G3.1] (ab2790) in 1x HBSS + 0.02% TritonX-100 + 1.5% FBS for 3 hours at 25°C. A polyclonal goat anti-mouse IgG Alexa Fluor® 488 was used as the secondary antibody at 1/1000 dilution. DAPI nuclear stainining.

    See Abreview

  • All lanes : Anti-Hsp27 antibody [G3.1] (ab2790) at 1/500 dilution

    Lane 1 : MCF7 whole cell lysate
    Lane 2 : MDA-MB-231 whole cell lysate

    Lysates/proteins at 30 µg per lane.

    Secondary
    Goat anti-Rabbit IgG (H+L) at 1/10000 dilution
    Developed using the ECL technique

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


    Exposure time : 1 minute

    This image is courtesy of an abreview submitted by Chi Li, University of Louisville.

    See Abreview

  • Immunohistochemistry of paraffin embedded human breast carcinoma with ab2790 labeling Hsp27 at 0.5μg/ml.

  • Immunohistochemistry of paraffin embedded human prostate carcinoma with ab2790 labeling Hsp27 at 0.5μg/ml.

     

     

  • Cells were fixed with 4% formaldehyde in PEM buffer. The coverslip was incubated in blocking buffer of 5% powdered milk in TBS-T plus 0.02% sodium azide for 1 hour at room temperature. Blocking buffer was removed and primary antibody was added at a dilution of 1/200 and incubated overnight at 4 degrees celsius. The coverslips were then washed 4-5 times with blocking buffer for 5 minutes. Secondary antibody, goat anti-rabbit Alexa 594 (ab150080), was added at a dilution of 1/1000 and incubated at room temperature for one hour. From this point on coverslips were covered with foil to protect them from light. They were washed 5 times with TBS-T and then one time with PEM, for 5 minutes each wash. The coverslips were fixed 10-30 minutes in 4% formaldehyde in PEM buffer, then washed 3 times with PEM buffer for 5 minutes. 0.1M ammonium chloride in PEM buffer was added for 10 minutes to quench auto-florescence, and then slips were washed 2 times for 5 minutes in PEM followed by 3 washes for 5 minutes in

  • Anti-Hsp27 antibody [G3.1] (ab2790) at 1/1000 dilution + Mouse whole brain tissue lysate. at 10 µg

    Secondary
    An HRP-conjugated Goat polyclonal. at 1/10000 dilution
    Developed using the ECL technique

    Observed band size : 24 kDa (why is the actual band size different from the predicted?)


    Exposure time : 5 minutes

    This image is courtesy of an Abreview submitted by Brian Hitt

    Blocking Step: 5% Milk for 1 hour at 25°C.
    Gel Running Conditions: Reduced, Denaturing Bis-tris 4-12%

    See Abreview

  • Overlay histogram showing HeLa cells (ab150035) stained with ab2790 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum (ab7481)  / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab2790, 1/100 dilution) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.

  • ab2790 staining Hsp27 in A549 lung cancer cells by Flow Cytometry. Cells were harvested in trypsin, fixed with paraformaldehyde and permeabilized with 0.2% Triton X-100. The sample was incubated with the primary antibody (1/50 in 1x HBSS + 0.02% Triton X-100 + 1.5% FBS) for 3 hours at 25°C. An Alexa Fluor® 488-conjugated goat anti-mouse IgG polyclonal (1/2000) was used as the secondary antibody. Gating Strategy: No gating.

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  • Flow Cytometry analysis of mouse lewis lung carcinoma cells labeling Hsp27 with ab2790. Cells were harvested in trypsin, then fixed with praformaldehyde and permeabilized with 0.2% Triton X-100. Followed by incubation with Anti-Hsp27 antibody [G3.1] (ab2790)  at 1/200 dilution in 1x HBSS + 0.02% Triton X-100 + 1.5% FBS for 3 hours at 25°C. A polyclonal goat anti-mouse IgG Alexa Fluor® 488 secondary antibody was used at 1/2000.

    See Abreview

Anti-Hsp27 antibody [G3.1] (ab2790)参考文献

This product has been referenced in:
  • 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 . Read more (PubMed: 27301321) »
  • White NM  et al. Quantitative proteomic analysis reveals potential diagnostic markers and pathways involved in pathogenesis of renal cell carcinoma. Oncotarget 5:506-18 (2014). WB ; Human . Read more (PubMed: 24504108) »

See all 16 Publications for this product

Product Wall

Application Immunocytochemistry/ Immunofluorescence
Sample Chinese Hamster Cell (Chinese hamster ovary cell)
Permeabilization Yes - 0.2% TritonX-100
Specification Chinese hamster ovary cell
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1.5% · Temperature: 25°C
Fixative Paraformaldehyde
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提交于 Feb 02 2016

Application Flow Cytometry
Sample Mouse Cell (Mouse Lewis Lung Carcinoma cells)
Permeabilization Yes - 0.2% Triton X-100
Gating Strategy No gating
Specification Mouse Lewis Lung Carcinoma cells
Preparation Cell harvesting/tissue preparation method: trypsin
Sample buffer: Hanks’ Balanced Salt solution (HBSS)
Fixation Paraformaldehyde
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Verified customer

提交于 Jan 25 2016

Application IHC - Wholemount
Sample Mouse Tissue (Transplanted lung tumor section)
Specification Transplanted lung tumor section
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Verified customer

提交于 Dec 22 2015

Application IHC - Wholemount
Sample Human Tissue (Ovary adenocarcinoma)
Specification Ovary adenocarcinoma
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提交于 Dec 21 2015

Application Western blot
Sample African Green Monkey Cell lysate - whole cell (COS-7 cells)
Gel Running Conditions Reduced Denaturing (4-20% Bis-Tris)
Loading amount 100000 cells
Treatment 30 nM Bortezomib for 24 hours
Specification COS-7 cells
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
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Verified customer

提交于 Jul 03 2015

Application Flow Cytometry
Sample Human Cell (A549 lung cancer cells)
Permeabilization Yes - 0.2% Triton X-100
Gating Strategy No gating
Specification A549 lung cancer cells
Preparation Cell harvesting/tissue preparation method: trypsin
Sample buffer: Hanks’ Balanced Salt solution (HBSS)
Fixation Paraformaldehyde
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Verified customer

提交于 May 15 2015

Application Western blot
Sample Human Cell lysate - whole cell (breast cancer cell lines MCF7 and MDA-MB-231)
Gel Running Conditions Reduced Denaturing (4-20% Tris-glycine)
Loading amount 30 µg
Specification breast cancer cell lines MCF7 and MDA-MB-231
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
Username

Dr. Chi Li

Verified customer

提交于 Apr 24 2015

Application Immunocytochemistry/ Immunofluorescence
Blocking step Serum as blocking agent for 10 minute(s) · Concentration: 1.5% · Temperature: 25°C
Sample Human Cell (non small cell lung carcinoma NCI-H1299)
Specification non small cell lung carcinoma NCI-H1299
Permeabilization Yes - 0.2% TritonX-100
Fixative Formaldehyde
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提交于 Mar 18 2015

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Frozen sections)
Sample Mouse Tissue sections (Uterus)
Specification Uterus
Fixative Formaldehyde
Blocking step BSA as blocking agent for 30 minute(s) · Concentration: 3% · Temperature: 25°C
Username

Dr. Mukesh Jaiswal

Verified customer

提交于 Apr 12 2013

Thank you for your recent telephone enquiry.

I can confirm that both ab2790 and ab2787 antibodies are sold as ascites fluid. As discussed on the phone, unpurified antibodies, such as those sold as whole antiserum, ascites or tissue culture s...

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