製品の概要

  • 製品名Anti-Hsp27 antibody [G3.1]
    Hsp27 一次抗体 製品一覧
  • 製品の詳細
    Mouse monoclonal [G3.1] to Hsp27
  • アプリケーション適用あり: Flow Cyt, ELISA, IHC-Fr, ICC, ICC/IF, IHC-P, WB, IPmore details
  • 種交差性
    交差種: Mouse, Rat, Dog, Human, Non Human Primates
    交差が予測される動物種: Cow, Pig
  • 免疫原

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

  • ポジティブ・コントロール
    • Hela Cells.

製品の特性

アプリケーション

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.

アプリケーション Abreviews 特記事項
Flow Cyt 1/100. ab170190-Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
ELISA Use at an assay dependent concentration.
IHC-Fr 1/100.
ICC 1/500.
ICC/IF 1/50 - 1/500.
IHC-P 1/500. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

Use citrate buffer.

WB 1/1000.
IP Use at an assay dependent concentration.

Suggest 2μl

ターゲット情報

  • 機能Involved in stress resistance and actin organization.
  • 組織特異性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.
  • 関連疾患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.
  • 配列類似性Belongs to the small heat shock protein (HSP20) family.
  • 翻訳後修飾Phosphorylated in MCF-7 cells on exposure to protein kinase C activators and heat shock.
  • 細胞内局在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
  • 参照データベース
  • 別名
    • 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] 画像

  • Immunocytochemistry/Immunofluorescence analysis of Hsp27 shows staining in C6 cells. Hsp27 staining (green), F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were probed without (control) or with ab2790 (1:20) overnight at 4°C, washed with PBS and incubated with a DyLight-488 conjugated goat anti-mouse secondary antibody. Images were taken at 60X magnification.

  • Immunocytochemistry/Immunofluorescence analysis of Hsp27 shows staining in HeLa cells. Hsp27 staining (green), F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were probed without (control) or with ab2790 (1:200) overnight at 4°C, washed with PBS and incubated with a DyLight-488 conjugated goat anti-mouse secondary antibody. Images were taken at 60X magnification.

  • Immunocytochemistry/Immunofluorescence analysis of Hsp27 shows staining in U251 cells. Hsp27 staining (green), F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were probed without (control) or with ab2790 (1:200) overnight at 4°C, washed with PBS and incubated with a DyLight-488 conjugated goat anti-mouse secondary antibody. Images were taken at 60X magnification.

  • Immunocytochemistry/Immunofluorescence analysis of Hsp27 (green) in HeLa cells and negative control NIH3T3 cells. Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% BSA for 15 minutes at room temperature. Cells were probed with ab2790 (1:50) for at least 1 hour at room temperature, washed with PBS, and incubated with a DyLight 488 goat anti-mouse IgG secondary antibody (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. Images were taken at 20X magnification.

  • Immunocytochemistry/Immunofluorescence analysis of Hsp27 (green) in HeLa cells. Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% BSA for 15 minutes at room temperature. Cells were probed with (left panel) or without (right panel) ab2790 (1:100) for at least 1 hour at room temperature, washed with PBS, and incubated with a DyLight 488 goat-anti-mouse IgG secondary antibody (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. Images were taken at 20X magnification.

  • Immunoprecipitation of Hsp27 was performed on HeLa cells. Antigen:antibody complexes were formed by incubating 500µg whole cell lysate with 2 µl of ab2790 overnight on a rocking platform at 4°C. The immune complexes were captured on 50µl Protein A/G agarose, washed extensively, and eluted with buffer. HeLa cell lysate (25µg) was loaded as a positive control. 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 ab2790 (1:1000) overnight rotating at 4°C, washed in TBST, and probed with IP detection reagent-HRP (1:1000) for at least one hour. Chemiluminescent detection was performed.

  • Western blot analysis of Hsp27 was performed by loading 50µg of the indicated whole cell lysates per well onto a 4-20% Tris-HCl polyacrylamide gel. Proteins were transferred to a PVDF membrane and blocked with 5% BSA/TBST for at least 1 hour. The membrane was incubated with ab2790 (1:1000) overnight at 4°C on a rocking platform, washed in TBS-0.1%Tween 20, and incubated with a goat anti-mouse IgG-HRP secondary antibody (1:20,000) for at least 1 hour. Chemiluminescent detection was performed.

  • 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

  • Immunohistochemistry was performed on both normal and cancer biopsies of deparaffinized Human tonsil tissue tissues. To expose target proteins heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1:500 with a mouse monoclonal antibody recognizing Heat Shock Protein 27 ab2790 or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.

  • Immunohistochemistry was performed on both normal and cancer biopsies of deparaffinized Human breast carcinoma tissues. To expose target proteins heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1:500 with a mouse monoclonal antibody recognizing Heat Shock Protein 27 ab2790 or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.

  • Immunohistochemistry was performed on both normal and cancer biopsies of deparaffinized Human colon carcinoma tissues. To expose target proteins heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1:200 with a mouse monoclonal antibody recognizing Heat Shock Protein 27 ab2790 or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.

  • 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.

    See Abreview

Anti-Hsp27 antibody [G3.1] (ab2790) 使用論文

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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投稿 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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投稿 Jan 25 2016

Application IHC - Wholemount
Sample Mouse Tissue (Transplanted lung tumor section)
Specification Transplanted lung tumor section
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投稿 Dec 22 2015

Application IHC - Wholemount
Sample Human Tissue (Ovary adenocarcinoma)
Specification Ovary adenocarcinoma
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投稿 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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投稿 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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投稿 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
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Chi Li

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投稿 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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投稿 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
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Dr. Mukesh Jaiswal

Verified customer

投稿 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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