Key features and details
- Rabbit polyclonal to Hsp27 (phospho S85)
- Suitable for: ICC, IP, WB
- Reacts with: Rat, Human
- Isotype: IgG
存放说明Shipped 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.
存储溶液Preservative: 0.05% Sodium azide
Constituents: PBS, 3% BSA
Concentration information loading...
纯度Immunogen 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 at an assay dependent concentration.|
|IP||Use at an assay dependent concentration.
|WB||Use a concentration of 0.25 - 2 µg/ml. Detects a band of approximately 27 kDa.|
功能Involved in stress resistance and actin organization.
组织特异性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.
疾病相关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.
序列相似性Belongs to the small heat shock protein (HSP20) family.
翻译后修饰Phosphorylated in MCF-7 cells on exposure to protein kinase C activators and heat shock.
细胞定位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
- Heat shock 27kDa protein antibody
- 28 kDa heat shock protein antibody
- CMT2F antibody
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
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.
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.
ab5594 被引用在 8 文献中.
- 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 . 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 . PubMed: 27301321
- Kierulf-Lassen C et al. No Effect of Remote Ischemic Conditioning Strategies on Recovery from Renal Ischemia-Reperfusion Injury and Protective Molecular Mediators. PLoS One 10:e0146109 (2015). PubMed: 26720280
- Carlsen I et al. Quercetin attenuates cyclooxygenase-2 expression in response to acute ureteral obstruction. Am J Physiol Renal Physiol 308:F1297-305 (2015). PubMed: 25810437
- Vidyasagar A et al. Tubular expression of heat-shock protein 27 inhibits fibrogenesis in obstructive nephropathy. Kidney Int 83:84-92 (2013). WB . PubMed: 22971995
- Jain S et al. Expression of phosphorylated heat shock protein 27 during corneal epithelial wound healing. Cornea 31:820-7 (2012). PubMed: 22262220
- Vidyasagar A et al. HSP27 is involved in the pathogenesis of kidney tubulointerstitial fibrosis. Am J Physiol Renal Physiol 295:F707-16 (2008). WB ; Rat . PubMed: 18596079
- Kawano F et al. Role(s) of nucleoli and phosphorylation of ribosomal protein S6 and/or HSP27 in the regulation of muscle mass. Am J Physiol Cell Physiol 293:C35-44 (2007). WB ; Rat . PubMed: 17182729