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  1. Link

    hrp-hsp27-antibody-epr5477-ab194079.pdf

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Signal Transduction Protein Trafficking Chaperones Heat Shock Proteins
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Validated using a knockout cell lineRecombinantRabMAb

Recombinant HRP Anti-Hsp27 antibody [EPR5477] (ab194079)

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  • SDS
  • Certificate of Compliance
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Western blot - HRP Anti-Hsp27 antibody [EPR5477] (ab194079)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - HRP Anti-Hsp27 antibody [EPR5477] (ab194079)
  • Western blot - HRP Anti-Hsp27 antibody [EPR5477] (ab194079)
  • HRP Anti-Hsp27 antibody [EPR5477] (ab194079)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • HRP Rabbit monoclonal [EPR5477] to Hsp27
  • Suitable for: IHC-P, WB
  • Knockout validated
  • Reacts with: Human
  • Conjugation: HRP

Conjugates logo Related conjugates and formulations

Alexa Fluor® 594 Alexa Fluor® 647 Carrier Free Unconjugated

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Anti-Hsp27 antibody [EPR5477] - BSA and Azide free (ab229442)

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Overview

  • Product name

    HRP Anti-Hsp27 antibody [EPR5477]
    See all Hsp27 primary antibodies
  • Description

    HRP Rabbit monoclonal [EPR5477] to Hsp27
  • Host species

    Rabbit
  • Conjugation

    HRP
  • Tested applications

    Suitable for: IHC-P, WBmore details
  • Species reactivity

    Reacts with: Human
    Predicted to work with: Mouse, Rat
  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: HeLa cell lysate. IHC: normal human skeletal muscle tissue.
  • General notes

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

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

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Store at -20°C. Avoid freeze / thaw cycle. Store In the Dark.
  • Storage buffer

    pH: 7.40
    Preservative: 0.1% Proclin 300 Solution
    Constituents: 30% Glycerol (glycerin, glycerine), 1% BSA, PBS
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR5477
  • Isotype

    IgG
  • Research areas

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

Associated products

  • Alternative Versions

    • Anti-Hsp27 antibody [EPR5477] (ab109376)
    • Alexa Fluor® 647 Anti-Hsp27 antibody [EPR5477] (ab194078)
    • Alexa Fluor® 594 Anti-Hsp27 antibody [EPR5477] (ab215328)
    • Anti-Hsp27 antibody [EPR5477] - BSA and Azide free (ab229442)
  • Isotype control

    • HRP Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab199507)
  • Recombinant Protein

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

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab194079 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
IHC-P
1/500. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

ab199507 - Rabbit monoclonal IgG (HRP), is suitable for use an as isotype control with this antibody.

WB
1/5000. Detects a band of approximately 27 kDa (predicted molecular weight: 23 kDa).
Notes
IHC-P
1/500. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

ab199507 - Rabbit monoclonal IgG (HRP), is suitable for use an as isotype control with this antibody.

WB
1/5000. Detects a band of approximately 27 kDa (predicted molecular weight: 23 kDa).

Target

  • Function

    Involved in stress resistance and actin organization.
  • Tissue specificity

    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.
  • Involvement in disease

    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.
  • Sequence similarities

    Belongs 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 localization

    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
  • Database links

    • Entrez Gene: 3315 Human
    • Entrez Gene: 15507 Mouse
    • Entrez Gene: 24471 Rat
    • Omim: 602195 Human
    • SwissProt: P04792 Human
    • SwissProt: P14602 Mouse
    • SwissProt: P42930 Rat
    • Unigene: 520973 Human
    • Unigene: 13849 Mouse
    • Unigene: 3841 Rat
    see all
  • 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

Images

  • Western blot - HRP Anti-Hsp27 antibody [EPR5477] (ab194079)
    Western blot - HRP Anti-Hsp27 antibody [EPR5477] (ab194079)
    All lanes : HRP Anti-Hsp27 antibody [EPR5477] (ab194079) at 1/5000 dilution

    Lane 1 : Wild-type HAP1 whole cell lysate
    Lane 2 : HSPB1 (Hsp27) knockout HAP1 whole cell lysate

    Lysates/proteins at 20 µg per lane.

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


    Exposure time: 10 seconds


    ab194079 was shown to specifically react with Hsp27 in wild-type HAP1 cells as signal was lost in HSPB1 (Hsp27) knockout cells. Wild-type and HSPB1 (Hsp27) knockout samples were subjected to SDS-PAGE. Ab194079 and ab184095 (Mouse monoclonal [mAbcam 9484] to GAPDH - Loading Control (Alexa Fluor® 680) loading control) were incubated overnight at 4°C at 1/5000 dilution and 1/1000 dilution respectively. The loading control was imaged using the Licor Odyssey CLx prior to blots being developed with ECL technique.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - HRP Anti-Hsp27 antibody [EPR5477] (ab194079)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - HRP Anti-Hsp27 antibody [EPR5477] (ab194079)

    IHC image of Hsp27 staining in a section of formalin-fixed paraffin-embedded normal human skeletal muscle tissue*, performed on a Leica BOND. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20mins. The section was then incubated with ab194079 at 1/500 dilution, for 15 mins at room temperature. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset negative control image is taken from an identical assay without primary antibody.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

    *Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre

  • Western blot - HRP Anti-Hsp27 antibody [EPR5477] (ab194079)
    Western blot - HRP Anti-Hsp27 antibody [EPR5477] (ab194079)
    HRP Anti-Hsp27 antibody [EPR5477] (ab194079) at 1/5000 dilution + HeLa whole cell lysate (ab150035) at 10 µg

    Developed using the ECL technique.

    Performed under reducing conditions.

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


    Exposure time: 20 seconds


    This blot was produced using a 4-12% Bis-tris gel under the MES buffer system. The gel was run at 200V for 35 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 2% Bovine Serum Albumin before being incubated with ab194079 overnight at 4°C. Antibody binding was visualised using ECL development solution ab133406.

  • HRP Anti-Hsp27 antibody [EPR5477] (ab194079)
    HRP Anti-Hsp27 antibody [EPR5477] (ab194079)

Protocols

  • Western blot protocols
  • Immunohistochemistry protocols

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

Certificate of Compliance

To download a Certificate of Compliance, please enter your Lot number below:

References (0)

Publishing research using ab194079? Please let us know so that we can cite the reference in this datasheet.

ab194079 has not yet been referenced specifically in any publications.

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