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ELISA - ASK1 (phospho S83) antibody (ab34650)
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Western blot - ASK1 (phospho S83) antibody (ab34650)
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Product Name
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ASK1 (phospho S83) antibody
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See all ASK1 antibodies (13)...
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Product type
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Primary antibodies
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Description
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Rabbit polyclonal to ASK1 (phospho S83)
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Immunogen
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Synthetic peptide: ATRGR GSSVGGG conjugated to KLH, corresponding to amino acids 76-87 of Human ASK1
BLAST 'ATRGR GSSVGGG' with  or 
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Reacts with
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Hu
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Does not react with
(species key)
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Ms
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Specificity
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ab34650 recognises ASK-1 (apoptosis signal-regulating kinase 1 - also referred to as MEK Kinase-5 or MAPKKK5).
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Tested applications
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ELISA, WB
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Abreviews
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Application notes
(see key)
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Recommended dilutions
ELISA: 1/5000 - 1/10000.
WB: 1/1000. Detects a band of approximately 155 kDa.
Not yet tested in other applications.
Optimal dilutions/concentrations should be determined by the end user.
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Positive control
(see definition)
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SW1353 whole cell lysate
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Cellular localization
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Cytoplasmic
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Research areas
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Cardiovascular >> Heart >> Hypertrophy >> Other
Cardiovascular >> Heart >> Apoptosis
Cell Biology >> Apoptosis >> Intracellular >> Kinases
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Relevance
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Mitogen activated protein (MAP) kinase cascades are activated in response to various extracellular stimuli including cytokines, growth factors and environmental stresses. A novel MAP kinase kinase kinase (MAPKKK) was recently identified and designated ASK1 (for apoptosis signal-regulating kinase 1). ASK1 activates two different subgroups of MAPKK, MKK4 and MKK6, which in turn activate c-Jun N terminal kinase (JNK) and p38 MAP kinase, respectively. ASK1 is activated by TNFR and Fas through the interaction with members of the TRAF family and Fas associated protein Daxx. Overexpression of ASK1 induces apoptotic cell death, and a catalytically inactive form of ASK1 inhibits TNF alpha-induced apoptosis. ASK1 is expressed in variety of human and mouse tissues.
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Database links
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The links below go to external sites and will open in a new browser window |
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Raised in
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Rabbit
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Clonality
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Polyclonal
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Isotype
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IgG
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Purity
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IgG fraction
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Storage buffer
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Preservative: 0.01% Sodium Azide
Constituents: 0.15M Sodium chloride, 0.02M Potassium phosphate. pH 7.2 Material safety datasheet (MSDS) for this product:
Sodium Azide MSDS
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Purification notes
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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 storage buffer.
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Form
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Liquid
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Concentration
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2.000 mg/ml
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Storage instructions
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Store at +4°C short term (1-2 weeks). Aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
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At Abcam, we have one centralized database to hold all of our product information, so that everything we know about this ASK1 (phospho S83) antibody is on this datasheet. But please do contact us if you would like any reassurance! |
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See below for ASK1 (phospho S83) antibody images, references, products related to ab34650 and other tools.
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ASK1 (phospho S83) antibody images:
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 ELISA - ASK1 (phospho S83) antibody (ab34650)
ab34650, tested
against BSA conjugates of non-phospho and
phospho forms of immunizing peptide by ELISA. Each
well was coated with 0.1 mg of conjugate. The
starting dilution of antibody was 1:1,000 and
each point on the X-axis represents a 2-fold
dilution. HRP conjugated Gt-a-Rabbit IgG H&L
and TMB substrate were used for detection.
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Western blot - ASK1 (phospho S83) antibody (ab34650)
Ab34650, generated by immunization with phospho peptide coupled to KLH, was tested by immunoblot against lysates of Cos-7 cells after transient transfection, separately, 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 uL of 1x SDS-sample buffer, heated
at 96°C for 5', and vortexed for 30 sec. Samples (10 uL 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 10 ug/mL mouse
anti-HA antibody for 45'. After 5X washes with TTBS, reaction with ALP rabbit anti-mouse IgG at 200 ng/mL proceeded for 45' following again by washing as before. The blot was developed using BCIP/NBT. This blot demonstrates both recombinant transfections were successfully over-expressed in the Cos-7 cells.
In panel b) all samples were incubated with a 1:1,000 dilution of ab34650 for 45'. The antibody was pre-incubated with non-phospho peptide prior to membrane incubation. After 5X washes with TTBS, reaction with HRP goat anti-rabbit IgG at 10 ng/mL proceeded for 45' following again by washing as before. The membrane was
processed as before. 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 a 1:1,000 dilution of ab34650 as before except the antibody was preincubated with phospho peptide prior to membrane incubation. No staining is observed after phospho peptide blocking occurs.
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Search PubMed (MEDLINE) for references to ASK1
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ASK1 (phospho S83) antibody - more information
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