Overview

  • Product name
    Mouse Hsp27 ELISA Kit
  • Detection method
    Colorimetric
  • Precision
    Intra-assay
    Sample n Mean SD CV%
    Cell extract 3 3.2%
    Inter-assay
    Sample n Mean SD CV%
    Cell extract 8 2.3%
  • Sample type
    Cell culture extracts
  • Assay type
    Sandwich (quantitative)
  • Sensitivity
    72 pg/ml
  • Range
    0.23 ng/ml - 15 ng/ml
  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Cell culture extracts 105 104% - 106%

  • Assay time
    1h 30m
  • Assay duration
    One step assay
  • Species reactivity
    Reacts with: Mouse
  • Product overview

    Hsp27 in vitro SimpleStep ELISA® (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of mouse Hsp27 protein in cell extracts.


    The SimpleStep ELISA® employs an affinity tag labeled capture antibody and a reporter conjugated detector antibody which immunocapture the sample analyte in solution. This entire complex (capture antibody/analyte/detector antibody) is in turn immobilized via immunoaffinity of an anti-tag antibody coating the well. To perform the assay, samples or standards are added to the wells, followed by the antibody mix. After incubation, the wells are washed to remove unbound material. TMB substrate is added and during incubation is catalyzed by HRP, generating blue coloration. This reaction is then stopped by addition of Stop Solution completing any color change from blue to yellow. Signal is generated proportionally to the amount of bound analyte and the intensity is measured at 450 nm. Optionally, instead of the endpoint reading, development of TMB can be recorded kinetically at 600 nm.

  • Notes

    Heat shock proteins (HSPs) are molecular chaperones that facilitate the proper folding and function of proteins within the cell. Heat shock protein 27 (Hsp27), also known as heat shock protein beta-1 (HspB1) is a 209 amino acid protein encoded by the HSPB1 gene. Hsp27 a member of the sHsp (small heat shock protein) group, whose functions are chaperone activity, thermotolerance, inhibition of apoptosis, and regulation of cellular development and differentiation. Hsp70 member proteins may assist inhibition of apoptosis by acting on the caspase-dependent pathway and against agents such as tumor necrosis factor-alpha, staurosporine, and doxorubicin. Hsp27 can form oligomers depending on the phosphorylation state of the protein and during heat shock. The oligomers are connected to the chaperone activity, while dimers have limited activity. Hsp27 interacts with actin and intermediate filaments, which provides protection to actin filaments from fragmentation.

  • Tested applications
    Suitable for: Sandwich ELISAmore details
  • Platform
    Pre-coated microplate (12 x 8 well strips)

Properties

  • Storage instructions
    Store at +4°C. Please refer to protocols.
  • Components 1 x 96 tests
    10X Mouse Hsp27 Detector Antibody 1 x 600µl
    10X Wash Buffer PT (ab206977) 1 x 20ml
    50X Cell Extraction Enhancer Solution (ab193971) 1 x 1ml
    5X Cell Extraction Buffer PTR (ab193970) 1 x 10ml
    Antibody Diluent CPR 1 x 6ml
    Mouse Hsp27 Capture Antibody Lyophilized 1 vial
    Mouse Hsp27 Lyophilized Recombinant Protein 2 vials
    Plate Seals 1 unit
    Sample Diluent NS 1 x 12ml
    SimpleStep Pre-Coated 96-Well Microplate (ab206978) 1 unit
    Stop Solution 1 x 12ml
    TMB Development Solution 1 x 12ml
  • Research areas
  • 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.
  • Information by UniProt
  • Alternative names
    • Heat shock 27kDa protein
    • 28 kDa heat shock protein
    • CMT2F
    • DKFZp586P1322
    • epididymis secretory protein Li 102
    • Estrogen regulated 24 kDa protein
    • Estrogen-regulated 24 kDa protein
    • Heat shock 25kDa protein 1
    • Heat shock 27 kDa protein
    • Heat shock 27kD protein 1
    • Heat shock 27kDa protein 1
    • Heat shock 28kDa protein 1
    • Heat Shock Protein 27
    • Heat shock protein beta 1
    • Heat shock protein beta-1
    • heat shock protein family B (small) member 1
    • HEL-S-102
    • HMN2B
    • HS.76067
    • Hsp 25
    • HSP 27
    • Hsp 28
    • Hsp B1
    • Hsp25
    • HSP27
    • Hsp28
    • HspB1
    • HSPB1_HUMAN
    • SRP27
    • Stress responsive protein 27
    • Stress-responsive protein 27
    see all
  • Database links

Applications

Our Abpromise guarantee covers the use of ab216949 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
Sandwich ELISA Use at an assay dependent concentration.

Images

  • Background-subtracted data values (mean +/- SD) are graphed.

  • The concentrations of Hsp27 were measured in duplicate and interpolated from the Hsp27 standard curve and corrected for sample dilution. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Hsp27 concentration was determined to be 540 ng/mg in C2C12 cell extract.

  • The concentrations of Hsp27 were measured in duplicate and interpolated from the Hsp27 standard curve and corrected for sample dilution. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Hsp27 concentrations were determined to be 680 ng/mg in mock-treated, 1470 ng/mg in heat-shocked, and 700 ng/mg in untreated C2C12 cell extract.

Protocols

References

ab216949 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

There are currently no Customer reviews or Questions for ab216949.
Please use the links above to contact us or submit feedback about this product.

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

Sign up