Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [Y175] to Hsp27 (phospho S78)
- Suitable for: WB, IHC-P, ICC/IF, Dot blot
- Reacts with: Human
Product nameAnti-Hsp27 (phospho S78) antibody [Y175]
See all Hsp27 primary antibodies
DescriptionRabbit monoclonal [Y175] to Hsp27 (phospho S78)
Specificityab32501 recognises Hsp27 (phospho S78). The antibody will detect Src phosphorylation on Serine 78.
Tested applicationsSuitable for: WB, IHC-P, ICC/IF, Dot blotmore details
Unsuitable for: Flow Cyt or IP
Species reactivityReacts with: Human
Synthetic phospho-peptide corresponding to residues surrounding Serine 78 of human HSP27
- HeLa cells, human breast carcinoma
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
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
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.
Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.
We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.
In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.
We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.
Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.
Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.
Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
Storage bufferpH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 49% PBS, 50% Glycerol (glycerin, glycerine), 0.05% BSA
Concentration information loading...
Our Abpromise guarantee covers the use of ab32501 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||1/2000. Predicted molecular weight: 23 kDa.|
|IHC-P||Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.|
|ICC/IF||1/250 - 1/500.|
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.
modificationsPhosphorylated 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
- Heat shock 27kDa protein antibody
- 28 kDa heat shock protein antibody
- CMT2F antibody
All lanes : Anti-Hsp27 (phospho S78) antibody [Y175] (ab32501) at 1/5000 dilution
Lane 1 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysates
Lane 2 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysates, treated with anisomycin.
Lane 3 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysates, treated with anisomycin. Then the membrane was incubated with phosphatase.
Lysates/proteins at 15 µg per lane.
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
Predicted band size: 23 kDa
Observed band size: 27 kDa why is the actual band size different from the predicted?
Blocking buffer 5% NFDM/TBST
Diluting buffer 5% NFDM/TBST
Secondary Ab at 1/20,000 dilution
ab32501 staining Hsp27 in HeLa cells by Immunocytochemistry. Tissue was fixed with 4% paraformaldehyde. Samples were incubated with primary antibody (4.5 μg/ml). ab150077 AlexaFluor®488 Goat anti-Rabbit (1/1000) was used as the secondary antibody. Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) at 2.5 μg/ml and DAPI were used as counter stains.
Confocal image showing the expression was increased after treatment with anisomycin (25ug/ml for 30min) and then decreased after treatment with the Lambda Protein Phosphatase 31℃ for 2h
Dot Blot analysis on immunogen phospho-peptide (A) and non-phospho peptide (B) using ab32501 at dilution 1/2000.
Immunohistochemical analysis of Hsp27 phospho S78 expression in paraffin embedded human breast carcinoma using 1/250 ab32501.
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Immunofluorescent analysis of Hsp27 phospho S78 expression in HeLa cells using 1/250 ab32501.
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
ab32501 has been referenced in 5 publications.
- Drexler R et al. Significance of unphosphorylated and phosphorylated heat shock protein 27 as a prognostic biomarker in pancreatic ductal adenocarcinoma. J Cancer Res Clin Oncol N/A:N/A (2020). IHC-P ; Human . PubMed: 32200459
- Li KC et al. Reduced expression of HSP27 following HAD-B treatment is associated with Her2 downregulation in NIH:OVCAR-3 human ovarian cancer cells. Mol Med Rep 12:3787-94 (2015). PubMed: 26044344
- de la Cuesta F et al. A proteomic focus on the alterations occurring at the human atherosclerotic coronary intima. Mol Cell Proteomics 10:M110.003517 (2011). IHC-Fr ; Human . PubMed: 21248247
- Shiryaev A et al. Distinct roles of MK2 and MK5 in cAMP/PKA- and stress/p38MAPK-induced heat shock protein 27 phosphorylation. J Mol Signal 6:4 (2011). WB . PubMed: 21575178
- Ning Y et al. IFNgamma restores breast cancer sensitivity to fulvestrant by regulating STAT1, IFN regulatory factor 1, NF-kappaB, BCL2 family members, and signaling to caspase-dependent apoptosis. Mol Cancer Ther 9:1274-85 (2010). WB ; Human . PubMed: 20457620