When tested by Western blotting on TNF alpha stimulated HeLa lysates the antibody detects a single clean band of about 27kD. This band is only blocked by the phospho peptide immunogen and not by an equivalent non-phospho peptide or by a generic phospho peptide. This confirms that the antibody is specific for phospho S82 of Hsp27.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
1/1000. Detects a band of approximately 27 kDa (predicted molecular weight: 27 kDa).
Involved in stress resistance and actin organization.
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.
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.
Belongs to the small heat shock protein (HSP20) family.
Phosphorylated in MCF-7 cells on exposure to protein kinase C activators and heat shock.
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.
heat shock protein family B (small) member 1 antibody
Hsp 25 antibody
HSP 27 antibody
Hsp 28 antibody
Hsp B1 antibody
Stress responsive protein 27 antibody
Stress-responsive protein 27 antibody
Western blot - Hsp27 (phospho S82) antibody (ab17937)
Predicted band size : 27 kDa
Western blot using ab17937 on HeLa cell lysate.
Lane 1: Unstimulated HeLa lysate
Lane 2: TNF-α Stimulated HeLa lysate
Lane 3: TNF-α Stimulated HeLa lysate blocked with non-phospho peptide (equivalent to immunogen sequence)
Lane 4: TNF-α Stimulated HeLa lysate blocked with generic phospho-serine peptide.
Lane 5: TNF-α Stimulated HeLa lysate blocked with phosphopeptide immunogen.
10-30 µg of cell lysate can be loaded when using similar lysates with this antibody. Samples were run using SDS-PAGE on a 10% polyacrylamide gel and transferred to PVDF. Membranes were blocked with a 5% BSA-TBST buffer for one hour at room temperature, then incubated with ab17937 for one hour at room temperature in 3% BSA-TBST buffer, following prior incubation with blocking peptides. After washing, membranes were incubated with goat F(ab')2 anti-rabbit IgG HRP conjugate in 3% BSA-TBST buffer, and bands were detected using the Pierce SuperSignal method.
Only the phospho peptide immunogen blocks the antibody, confirming its specificity for the phospho epitope S82.
Svensson KJ et al. Exosome uptake depends on ERK1/2-heat shock protein 27 signaling and lipid Raft-mediated endocytosis negatively regulated by caveolin-1. J Biol Chem288:17713-24 (2013).
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Wang YT et al. An informatics-assisted label-free quantitation strategy that depicts phosphoproteomic profiles in lung cancer cell invasion. J Proteome Res9:5582-97 (2010).
Read more (PubMed: 20815410) »