Recombinant Anti-Hsp27 (phospho S78) antibody [Y175] (ab32501)
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: ICC/IF, WB, IHC-P, Dot blot
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-Hsp27 (phospho S78) antibody [Y175]
See all Hsp27 primary antibodies -
Description
Rabbit monoclonal [Y175] to Hsp27 (phospho S78) -
Host species
Rabbit -
Specificity
ab32501 recognises Hsp27 (phospho S78). The antibody will detect Src phosphorylation on Serine 78.
This antibody does not react with mouse and rat species in the Western blot application.
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Tested applications
Suitable for: ICC/IF, WB, IHC-P, Dot blotmore details
Unsuitable for: Flow Cyt -
Species reactivity
Reacts with: Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: HeLa starved overnight then treated with 250 ng/ml anisomycin for 30 minutes whole cell lysate. IHH: Human breast cancer tissue sections. ICC/IF: HeLa treated with 25 ug/mL anisomycin for 30 min, then Lambda Protein Phosphatase 31 for 2 hours cells.
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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
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles. -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
Y175 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Isotype control
Applications
The Abpromise guarantee
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.
Application | Abreviews | Notes |
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ICC/IF |
1/50.
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WB |
1/5000. Predicted molecular weight: 23 kDa.
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IHC-P |
1/5000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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Dot blot |
1/2000.
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Notes |
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ICC/IF
1/50. |
WB
1/5000. Predicted molecular weight: 23 kDa. |
IHC-P
1/5000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
Dot blot
1/2000. |
Target
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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
modificationsPhosphorylated 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
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Database links
- Entrez Gene: 3315 Human
- Omim: 602195 Human
- SwissProt: P04792 Human
- Unigene: 520973 Human
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Alternative names
- Heat shock 27kDa protein antibody
- 28 kDa heat shock protein antibody
- CMT2F antibody
see all
Images
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All lanes : Anti-Hsp27 (phospho S78) antibody [Y175] (ab32501) at 1/5000 dilution (Purified)
Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate
Lane 2 : HeLa (Human cervix adenocarcinoma epithelial cell) starved overnight then treated with 250 ng/ml anisomycin for 30 minutes whole cell lysate
Lane 3 : HeLa (Human cervix adenocarcinoma epithelial cell) starved overnight then treated with 250 ng/ml anisomycin for 30 minutes whole cell lysate, and then the menbrane treated with Alkaline Phosphatase for 1 hour
Lysates/proteins at 15 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 23 kDa
Observed band size: 27 kDa why is the actual band size different from the predicted? -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human breast cancer tissue sections labelling Hsp27 with purified ab32501 at 1/5000 dilution (0.11 µg/ml). Heat mediated antigen retrieval was performed using Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). Tissue was counterstained with Hematoxylin. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) secondary antibody was used at 1/0 dilution. PBS instead of the primary antibody was used as the negative control. Positive staining on human breast cancer without alkaline phosphatase treatment (image A). No staining on human breast cancer with alkaline phosphatase treatment (image B). The immunostaining was performed on a Leica Biosystems BOND® RX instrument.
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Immunocytochemistry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) treated with 25 ug/mL anisomycin for 30 min, then Lambda Protein Phosphatase 31? for 2 hours cells labeling Hsp27 with purified ab32501 at 1/50 dilution (11.3 µg/mL). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1/200 (2.5 µg/mL). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1/1000 (2 µg/mL) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
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Dot Blot analysis on immunogen phospho-peptide (A) and non-phospho peptide (B) using ab32501 at dilution 1/2000.
Protocols
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.
Datasheets and documents
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SDS download
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Datasheet download
References (8)
ab32501 has been referenced in 8 publications.
- Coates MS et al. Pseudomonas aeruginosa induces p38MAP kinase-dependent IL-6 and CXCL8 release from bronchial epithelial cells via a Syk kinase pathway. PLoS One 16:e0246050 (2021). PubMed: 33524056
- Li Z et al. Spinal heat shock protein 27 participates in PDGFRβ-mediated morphine tolerance through PI3K/Akt and p38 MAPK signalling pathways. Br J Pharmacol 177:5046-5062 (2020). PubMed: 32559815
- 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
- Zhang Y et al. Heat Shock Protein 27 Regulates the Inflammatory Response of Intestinal Epithelial Cells by the Nuclear Factor-?B Pathway. Dig Dis Sci 65:3514-3520 (2020). PubMed: 32078087
- 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