Overview

  • Product nameAnti-Hsp27 antibody
    See all Hsp27 primary antibodies
  • Description
    Rabbit polyclonal to Hsp27
  • Tested applicationsSuitable for: WB, ICC, ICC/IF, IHC-Pmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human, Non Human Primates
    Predicted to work with: Cow, Dog, Pig
  • Immunogen

    Synthetic peptide corresponding to Human Hsp27 aa 10-21.
    Sequence:

    LLRGPSWDPFRC


    (Peptide available as ab39789)

  • Positive control
    • HeLa cell lysate.

Properties

Associated products

Applications

Our Abpromise guarantee covers the use of ab5579 in the following tested applications.

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

Application Abreviews Notes
WB 1/1000 - 1/2000. Detects a band of approximately 27 kDa.Can be blocked with Human Hsp27 peptide (ab39789).
ICC Use at an assay dependent concentration.
ICC/IF 1/50.
IHC-P Use a concentration of 4 µg/ml. Perform heat mediated antigen retrieval via the microwave method before commencing with IHC staining protocol.

Target

  • FunctionInvolved in stress resistance and actin organization.
  • Tissue specificityDetected 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.
  • Involvement in diseaseDefects 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.
  • Sequence similaritiesBelongs to the small heat shock protein (HSP20) family.
  • Post-translational
    modifications
    Phosphorylated in MCF-7 cells on exposure to protein kinase C activators and heat shock.
  • Cellular localizationCytoplasm. 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
  • Database links
  • Alternative names
    • 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 images

  • Immunocytochemistry/Immunofluorescence analysis of C6 cells labeling Hsp27 (green) with ab5579 at 1/100. F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue). Cells were fixed with formaldehyde and incubated with the primary antibody overnight at 4°C. A DyLight 488-conjugated secondary antibody was used. 60X magnification. Right - negative control.

  • Immunocytochemistry/Immunofluorescence analysis of HeLa cells labeling Hsp27 (green) with ab5579 at 1/200. F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue). Cells were fixed with formaldehyde and incubated with the primary antibody overnight at 4°C. A DyLight 488-conjugated secondary antibody was used. 60X magnification. Right - negative control.

  • Immunocytochemistry/Immunofluorescence analysis of MCF-7 cells labeling Hsp27 (green) with ab5579 at 1/200. F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue). Cells were fixed with formaldehyde and incubated with the primary antibody overnight at 4°C. A DyLight 488-conjugated secondary antibody was used. 60X magnification. Right - negative control.

  • Western blot analysis of various cell lines (50µg/lable whole cell lysates) labeling Hsp27 with ab5579 at 1/1000. A HRP-conjugated goat anti-rabbit IgG (1/20000) was used as the seondary antibody.

  • HeLa 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 1/250 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 minute

  • Immunohistochemistry was performed on both normal and cancer biopsies of deparaffinized human breast carcinoma tissue. 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 1/100 with a rabbit polyclonal antibody recognizing Heat Shock Protein 27 (ab5579) 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 skeletal muscle tissue. To expose target proteins, heat induced antigen retrieval was performed using 10mM sodium citrate (pH 6.0) 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 1/20 with a rabbit polyclonal antibody recognizing Heat Shock Protein 27 (ab5579) 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.

References for Anti-Hsp27 antibody (ab5579)

This product has been referenced in:
  • Choi SH  et al. MMP9 processing of HSPB1 regulates tumor progression. PLoS One 9:e85509 (2014). IHC-P ; Mouse . Read more (PubMed: 24465581) »
  • Svensson KJ  et al. Exosome uptake depends on ERK1/2-heat shock protein 27 signaling and lipid Raft-mediated endocytosis negatively regulated by caveolin-1. J Biol Chem 288:17713-24 (2013). Read more (PubMed: 23653359) »

See all 5 Publications for this product

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Thank you for contacting us. Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE" I hope this information is helpful to you. Please do not hesitate to contact us if you need any more adv...

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Rat Tissue lysate - whole (Spleen)
Specification Spleen
Blocking step Other as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: N/A
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Verified customer

Submitted Oct 18 2005

Thank you for your enquiry. We are going to send you a free of charge replacement vial very shortly. For your information, the new order number is: 36004. We apologize for any inconvenience caused and thank you for bringing this matter to our at...

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