The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
1/2000. Predicted molecular weight: 23 kDa.
Use at an assay dependent concentration.
1/250 - 1/500.
Use at an assay dependent concentration. PubMed: 21248247
Application notesIs unsuitable for Flow Cyt or IP.
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.
Post-translational 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.
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
Western blot - Anti-Hsp27 (phospho S78) antibody [Y175] (ab32501)
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.
Immunofluorescent analysis of Hsp27 phospho S78 expression in HeLa cells using 1/250 ab32501.
References for Anti-Hsp27 (phospho S78) antibody [Y175] (ab32501)
This product has been referenced in:
de la Cuesta F et al. A proteomic focus on the alterations occurring at the human atherosclerotic coronary intima. Mol Cell Proteomics10:M110.003517 (2011).
Read more (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 Signal6:4 (2011).
Read more (PubMed: 21575178) »