Overview

  • Product name
    Anti-Hsp27 antibody [G3.1]
    See all Hsp27 primary antibodies
  • Description
    Mouse monoclonal [G3.1] to Hsp27
  • Host species
    Mouse
  • Tested applications
    Suitable for: Flow Cyt, IHC-Fr, ICC/IF, IHC-P, WBmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Sheep, Chicken, Human, Non human primates
    Predicted to work with: Cow, Pig
  • Immunogen

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

  • Positive control
    • Hela Cells.

Properties

Applications

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.

Application Abreviews Notes
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.

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.
  • 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

Images

  • 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

  • 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.

     

     

  • 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


    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.

References

This product has been referenced in:
  • Sun P  et al. Foot-and-mouth disease virus capsid protein VP2 activates the cellular EIF2S1-ATF4 pathway and induces autophagy via HSPB1. Autophagy 14:336-346 (2018). Read more (PubMed: 29166823) »
  • San Gil R  et al. Using bicistronic constructs to evaluate the chaperone activities of heat shock proteins in cells. Sci Rep 7:2387 (2017). Read more (PubMed: 28539657) »
See all 29 Publications for this product

Customer reviews and Q&As

1-9 of 9 Abreviews or Q&A

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

Dr. Mukesh Jaiswal

Verified customer

Submitted Apr 12 2013

Question
Answer

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 supernatant will not have a concentration stated on the datasheet.

Antibody concentration is usually determined by protein assay, and serum / ascites / tissue culture supernatant will contain a lot of other proteins, which means the antibody quantification would not be accurate.

I can confirm that for ascites, concentration of antibody is known to very between 5 - 10 mg/ml.

I am sorry we are not able to provide an exact concentration on this occasion, but hope this information will be helpful to you. If you have any further questions, please do not hesitate to contact us.

Read More

Answer

Thank you for your enquiry and your interest in our products.

I can confirm that these two products are the same clone (therefore the clone number G3.1 is identical) so the immunogen sequence used to raise them was exactly the same. However, ab2790 is conjugated whilst ab115639 labelled to DyLight® 488.

If you need any further assistance in the future, please do not hesitate to contact me.

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (A549 cells)
Loading amount
50000 cells
Specification
A549 cells
Treatment
26 nM DMSO/bortezomib (BTZ) 24 hours
Gel Running Conditions
Reduced Denaturing (NuPage 4-12%)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Aug 15 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Mouse Tissue lysate - whole (Whole brain homogenate)
Loading amount
10 µg
Specification
Whole brain homogenate
Gel Running Conditions
Reduced Denaturing (Bis-tris 4-12%)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Mr. Brian Hitt

Verified customer

Submitted Jun 01 2010

Answer

Thank you for your enquiry. I can confirm that this product has been successfully used in IHC in the following article. Ye H et al. Proteomic based identification of manganese superoxide dismutase 2 (SOD2) as a metastasis marker for oral squamous cell carcinoma. Cancer Genomics Proteomics 5:85-94 (2008). WB, IHC-P; Human. PubMed: 18460737 http://www.ncbi.nlm.nih.gov/pubmed/18460737?dopt=Abstract Unfortunately, I was not able to locate the data demonstrating reactivity with mouse HSP27 at this time. Therefore, I will temporarily change the mouse reactivity from the list of approved species to "predicted to react" until we can produce the required data. However, I have run a sequence align and found that the mouse HSP27 sequence is strongly conserved within the human sequence (see attached file). Also, this is one of our oldest clones, and I there are no customer complaints for using this antibody in mouse. Therefore, we expect that it will work for your too.

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunoprecipitation
Sample
Human Cell lysate - whole cell (Colon cancer cell line HCT116)
Total protein in input
30 µg
Specification
Colon cancer cell line HCT116
Immuno-precipitation step
Protein A

Abcam user community

Verified customer

Submitted Oct 09 2006

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (Colon cancer cell line HCT116)
Loading amount
40 µg
Specification
Colon cancer cell line HCT116
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% in TBST

Abcam user community

Verified customer

Submitted Oct 03 2006

Question
Answer

Thank you for your enquiry. Regarding the HSP-27 antibody (ab2790), a sodium citrate heat mediated antigen retrieval method has been used with this product in IHC techniques (I have added this information to the datasheet for future reference). I would recommend adding a permeabilization step, such as Triton X-100 or saponin, as well. I am still looking into ab13494 for you, and will email again once I have more information.

Read More

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