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Anti-ASK1 pS83 (RABBIT) Antibody

ASK1 phospho S83 Antibody

     
  •  - Anti-ASK1 pS83 (RABBIT) Antibody ASR4387
    Western blot of anti-pS83 ASK1 antibodies shows specificity for phosphorylated human ASK1. Anti-pS83 (aa 76-87) antibody was tested by western blot against Cos-7 cell lysates after transient transfection with 1) vector only, 2) recombinant HA-ASK1, and 3) recombinant human HA-ASK1 where S83 was substituted with an alanine residue. Cells were lysed 24 h post-transfection in 200 µL of 1x SDS-sample buffer, heated at 96°C for 5', and vortexed for 30 sec. Samples (10 µL each) were separated on a 12% SDS-PAGE gel and transferred to PVDF (Millipore) followed by blocking for 45' using TTBS supplemented with 5% non-fat dry milk. All incubations were performed at room temperature. In panel a) all samples were incubated with anti-HA antibody. This blot demonstrates both recombinant transfections express rASK1. In panel b) all samples were incubated with anti-pS83 ASK1. Lane 2 shows strong specific staining of ASK1. Lane 3, where S83 was replaced with alanine, shows greatly diminished staining. In panel c) all samples were incubated with anti-pS83 ASK1 antibody as before except the antibody was pre-incubated with phospho peptide prior to membrane incubation. No staining is observed after phospho peptide blocking occurs.
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Product Information
Host Rabbit
Conjugate Unconjugated
Target Species Human
Reactivity Human
Clonality Polyclonal
Application
  • Applications Legend:
  • WB=Western Blot
  • IHC=Immunohistochemistry
  • IHC-P=Immunohistochemistry (Paraffin-embedded Sections)
  • IHC-F=Immunohistochemistry (Frozen Sections)
  • IF=Immunofluorescence
  • FC=Flow Cytopmetry
  • IC=Immunochemistry
  • ICC=Immunocytochemistry
  • E=ELISA
  • IP=Immunoprecipitation
  • DB=Dot Blot
  • CHIP=Chromatin Immunoprecipitation
  • FA=Fluorescence Assay
  • IEM=Immuno electron microscopy
  • EIA=Enzyme Immunoassay
WB, E, I, LCI
Application Note This phospho specific polyclonal antibody reacts human pS83 ASK1 and shows minimal reactivity by western blot, ELISA and competitive ELISA with non-phosphorylated ASK1. Although not tested, this antibody is likely functional in RIA, immunohistochemistry and immunoprecipitation. For immunoblotting a 1:1,000 dilution is recommended.  A 155 kDa band corresponding to human ASK-1 is detected.  Whole cell lysates from SW1353 can be used as a positive control.  For ELISA a 1:5,000 to 1:10,000 dilution is recommended.   Researchers should determine optimal titers for other applications.
Physical State Liquid (sterile filtered)
Buffer 0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2
Immunogen This purified antibody was prepared from rabbit serum after repeated immunizations with a KLH conjugated peptide corresponding to an internal region near amino acids 75-100 of human ASK-1 protein.
Preservative 0.01% (w/v) Sodium Azide
Additional Information
Gene ID 4217
Other Names 4217
Purity This product is an IgG fraction antibody purified from antiserum by a multi-step process which includes delipidation, salt fractionation and ion exchange chromatography followed by extensive dialysis against the buffer stated above. Assay by immunoelectrophoresis resulted in a single precipitin arc against anti-Rabbit Serum. No reaction was observed with ASK-1 from mouse sources. Reactivity with the kinase from other sources has not been determined.
Storage Condition Store vial at -20° C prior to opening. Aliquot contents and freeze at -20° C or below for extended storage. Avoid cycles of freezing and thawing. Centrifuge product if not completely clear after standing at room temperature. This product is stable for several weeks at 4° C as an undiluted liquid. Dilute only prior to immediate use.
Precautions NoteThis product is for research use only and is not intended for therapeutic or diagnostic applications.
Protein Information
Name MAP3K5
Synonyms ASK1, MAPKKK5, MEKK5
Function Serine/threonine kinase which acts as an essential component of the MAP kinase signal transduction pathway. Plays an important role in the cascades of cellular responses evoked by changes in the environment. Mediates signaling for determination of cell fate such as differentiation and survival. Plays a crucial role in the apoptosis signal transduction pathway through mitochondria-dependent caspase activation. MAP3K5/ASK1 is required for the innate immune response, which is essential for host defense against a wide range of pathogens. Mediates signal transduction of various stressors like oxidative stress as well as by receptor-mediated inflammatory signals, such as the tumor necrosis factor (TNF) or lipopolysaccharide (LPS). Once activated, acts as an upstream activator of the MKK/JNK signal transduction cascade and the p38 MAPK signal transduction cascade through the phosphorylation and activation of several MAP kinase kinases like MAP2K4/SEK1, MAP2K3/MKK3, MAP2K6/MKK6 and MAP2K7/MKK7. These MAP2Ks in turn activate p38 MAPKs and c-jun N-terminal kinases (JNKs). Both p38 MAPK and JNKs control the transcription factors activator protein-1 (AP-1).
Cellular Location Cytoplasm. Endoplasmic reticulum. Note=Interaction with 14-3-3 proteins alters the distribution of MAP3K5/ASK1 and restricts it to the perinuclear endoplasmic reticulum region
Tissue Location Abundantly expressed in heart and pancreas.
Research Areas
Citations (0)
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Background

ASK-1 (apoptosis signal-regulating kinase 1 - also referred to as MEK Kinase-5 or MAPKKK5) is a novel serine/threonine MAP kinase kinase kinase (MAPKKK) component of the mitogen -activated protein (MAP) cascade that is activated in response to extracellular stimuli by cytokines, growth factors and environmental stresses and other factors.   Overexpression of ASK-1 induces apoptotic cell death.  ASK-1 is expressed in a variety of human and mouse tissues.  The overall amino acid sequence identity between the mouse and human ASK1 is 91.9%.  ASK-1 interacts with CDKN1A (also known as p21, WAF1, CIP1).  Please refer to the reference list at the end of this document for further information.

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$ 475.00
Cat# ASR4387
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