Product nameAnti-Hsp27 (phospho S85) antibody
See all Hsp27 primary antibodies
DescriptionRabbit polyclonal to Hsp27 (phospho S85)
Tested applicationsSuitable for: ICC, IP, ICC/IF, WBmore details
Species reactivityReacts with: Rat, Human
Predicted to work with: Mouse, Cow, Dog, Pig, Xenopus laevis, Chinese hamster
Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
Storage bufferPreservative: 0.05% Sodium azide
Constituents: 99% PBS, 3% BSA
Concentration information loading...
PurityImmunogen affinity purified
Our Abpromise guarantee covers the use of ab5594 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|ICC||Use a concentration of 15 - 25 µg/ml.|
|IP||Use at an assay dependent concentration.
|ICC/IF||Use a concentration of 15 - 25 µg/ml.|
|WB||Use a concentration of 0.25 - 2 µg/ml. Detects a band of approximately 27 kDa.|
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
Immunocytochemistry/Immunofluorescence analysis of Hsp27 (phospho S85) (green) in HeLa cells either left untreated (left panel) or treated with 10uM Anisomysin (right panel) for 30 minutes. Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with ab5594 at 20ug/ml for at least 1 hour at room temperature, washed with PBS, and incubated with DyLight 488 goat anti-rabbit IgG secondary antibody at 1:400 for 30 minutes at room temperature. F-Actin (red) was stained with DyLight 554 Phalloidin and nuclei (blue) were stained with Hoechst 33342 dye. 20X magnification.
Immunoprecipitation of Hsp27 (phospho S85) was performed on HeLa cells treated with 10uM Anisomysin for 30 minutes. Antigen-antibody complexes were formed by incubating 500µg of whole cell lysate with 3µg of ab5594 overnight on a rocking platform at 4°C. The immune complexes were captured on 50µl Protein A/G Agarose, washed extensively, and eluted with Lane Marker Reducing Sample Buffer. HeLa cell lysate (50ug) was loaded as a positive control (left lane). Samples were resolved on a 4-20% Tris-HCl polyacrylamide gel, transferred to a PVDF membrane, and blocked with 5% BSA/TBST for at least 1 hour. The membrane was probed with ab5594 at 2ug/ml overnight rotating at 4°C, washed in TBST, and probed with Clean-Blot IP Detection Reagent at a dilution of 1:1000 for at least 1 hour. Chemiluminescent detection was performed using SuperSignal West Dura.
All lanes : Anti-Hsp27 (phospho S85) antibody (ab5594) at 2 µg/ml
Lane 1 : Hela cell lysate - untreated
Lane 2 : Hela cell lysate - treated with 10uM Anisomycin for 30 minutes
Lysates/proteins at 50 µg per lane.
All lanes : HRP-conjugated goat anti-rabbit IgG at 1/20000 dilution
Western blot analysis of Hsp27 (phospho S85) was performed by loading rat skeletal muscle tissue extracts on a SDS-PAGE gel. Proteins were transferred to a membrane probed with a Hsp27 (phospho S85) polyclonal antibody (ab5594) at a concentration of 0.25ug/ml followed by an anti-rabbit IgG-HRP secondary antibody. Detection was performed using a chemiluminescent substrate.
This product has been referenced in:
- Yamamoto Y et al. Pentose phosphate pathway activation via HSP27 phosphorylation by ATM kinase: A putative endogenous antioxidant defense mechanism during cerebral ischemia-reperfusion. Brain Res 1687:82-94 (2018). WB ; Rat . Read more (PubMed: 29510140) »
- Zhai W et al. A1 adenosine receptor attenuates intracerebral hemorrhage-induced secondary brain injury in rats by activating the P38-MAPKAP2-Hsp27 pathway. Mol Brain 9:66 (2016). Rat . Read more (PubMed: 27301321) »