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Primary Antibodies  >  Phospho Specific Antibodies
 

ASK-1 phospho S83 Antibody

Rabbit Polyclonal
Code

200-401-410
Size

200 µg
Price

$309.00
Availability

Ships next business day.
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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 ?L of 1x SDS-sample buffer, heated at 96?C for 5', and vortexed for 30 sec.  Samples (10 ?L 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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Synonyms: Apoptosis signal regulating kinase 1 antibody, ASK 1 antibody, ASK1 antibody, MAP/ERK kinase kinase 5 antibody, MAP3K5 antibody, MAPK/ERK kinase kinase 5 antibody

ASK-1 phospho S83 Antibody Properties

Description
Anti-ASK-1 pS83 (RABBIT) Antibody - 200-401-410
Host
Rabbit
Target Species
Human
Known Cross Reactivity
Human
Clonality
Polyclonal
Application
ELISA : 1:5,000 - 1:10,000
Western Blot : 1:1,000
Other Dilution: User Optimized
Physical State
Liquid (sterile filtered)
Shipping condition
Dry Ice
Format
IgG
Label
Concentration
1.0 mg/mL by UV absorbance at 280 nm
Buffer
0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2
Preservative
0.01% (w/v) Sodium Azide
Stabilizer
None
GeneName
MAP3K5
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Lot#  

ASK-1 phospho S83 Antibody Description

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.
Immunogen
This purified antibody was prepared from rabbit serum after repeated immunizations with a KLH conjugated peptide corresponding to amino acids 76-87 of human ASK-1 protein.
Immunogen Type
Peptide
Post Translational Modification
Phosphorylation
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.
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.
Purity/Specifity
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.
Disclaimer Note-General
This product is for research use only and is not intended for therapeutic or diagnostic applications. Please contact a technical service representative for more information. All products of animal origin manufactured by Rockland Immunochemicals are derived from starting materials of North American origin. Collection was performed in United States Department of Agriculture (USDA) inspected facilities and all materials have been inspected and certified to be free of disease and suitable for exportation. All properties listed are typical characteristics and are not specifications. All suggestions and data are offered in good faith but without guarantee as conditions and methods of use of our products are beyond our control. All claims must be made within 30 days following the date of delivery. The prospective user must determine the suitability of our materials before adopting them on a commercial scale. Suggested uses of our products are not recommendations to use our products in violation of any patent or as a license under any patent of Rockland Immunochemicals, Inc. If you require a commercial license to use this material and do not have one, then return this material, unopened to: Rockland Inc., P.O. BOX 326, Gilbertsville, Pennsylvania, USA.
General Reference
Morita, K. et al. (2001) Negative feedback regulation of ASK1 by protein phophatase 5 (PP5) in response to oxidative stress. EMBO J., 20, 6028-6036. Matsuzawa, A. and Ichijo, H. (2001) Molecular Mechanisms of the Decision between Life and Death: Regulation of Apoptosis by Apoptosis Signal-Regulating Kinase 1. J. Biochem., 130, 1-8. Zou, X. et al. (2001) The cell cycle-regulatory CDC25A phosphatase inhibits apoptosis signal-regulating kinase 1 (ASK1). Mol. Cell. Biol., 21, 4818-4828.
Specific Reference
Subramanyam M, Takahashi N, Hasegawa Y, et al. Inhibition of protein kinase Akt1 by apoptosis signal-regulating kinase-1 (ASK1) is involved in apoptotic inhibition of regulatory volume increase. J. Biol. Chem. 2010 Feb 26;285(9):6109-6117.
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Product Type
Primary Antibodies;
Applications
ELISA, WB, IHC, IF, FC
Reacts With
human, mouse, rat, chicken
Catalog Number
100-401-401

Product Type
Primary Antibodies;
Applications
ELISA, WB, IHC, IF, FC
Reacts With
human, mouse, rat, monkey
Catalog Number
200-301-268

Product Type
Primary Antibodies;
Applications
ELISA, WB
Reacts With
human, mouse, rat
Catalog Number
600-401-281

Product Type
Primary Antibodies;
Applications
ELISA, WB, IF
Reacts With
human, mouse
Catalog Number
600-401-B27

Product Label

 

Description
Unconjugated
Color
Conjugation Reference
Molecular Weight
Excitation Wavelength
Conjugation Chemistry