Product Name: ASK1-pS1033
Product Number: AB-PK524
Size: 25 µg      Price:89.00
      $US
Target Full Name: Apoptosis signal regulating protein-serine kinase 1

Target Alias: Apoptosis signal regulating kinase 1; Apoptosis signal-regulating kinase 1; ASK-1; Kinase ASK1; M3K5; MAP3K5; MAPK/ERK kinase kinase 5; MAPKKK5; MEK kinase 5; MEKK5; ENSG00000197442

Product Type Specific: Protein kinase phosphosite-specific antibody

Antibody Code: PK524

Antibody Target Type: Phosphosite-specific

Antibody Phosphosite: S1033

Protein UniProt: Q99683

Protein SigNET: Q99683

Antibody Type: Polyclonal

Antibody Host Species: Rabbit

Antibody Immunogen Source: Human ASK1 (MAP3K5) sequence peptide Cat. No.: PE-04ALQ95

Antibody Immunogen Sequence: APP(pS)PEE(bA)C

Antibody Immunogen Description: Corresponds to amino acid residues A1030 to E1036; After the kinase catalytic domain. This is the major in vivo phosphorylation site in ASK1.

Production Method: The immunizing peptide was produced by solid phase synthesis on a multipep peptide synthesizer and purified by reverse-phase hplc chromatography. Purity was assessed by analytical hplc and the amino acid sequence confirmed by mass spectrometry analysis. This peptide was coupled to KLH prior to immunization into rabbits. New Zealand White rabbits were subcutaneously injected with KLH-coupled immunizing peptide every 4 weeks for 4 months. The sera from these animals was applied onto an agarose column to which the immunogen peptide was thio-linked. Antibody was eluted from the column with 0.1 M glycine, pH 2.5. Subsequently, the antibody solution was neutralized to pH 7.0 with saturated Tris.This antibody was also subject to negative purification over phosphotyrosine-agarose.

Antibody Modification: Unconjugated. Contact KInexus if you are interest in having the antibody biotinylated or coupled with fluorescent dyes.

Antibody Concentration: 1 mg/ml

Storage Buffer: Phosphate buffered saline pH 7.4, 0.05% Thimerasol

Storage Conditions: For long term storage, keep frozen at -40°C or lower. Stock solution can be kept at +4°C for more than 3 months. Avoid repeated freeze-thaw cycles.

Product Use: Western blotting | Antibody microarray

Antibody Dilution Recommended: 2 µg/ml for immunoblotting

Antibody Potency: Medium immunoreactivity of a target-sized protein by Western blotting. Very strong immunoreactivity with immunogen peptide on dot blots.

Antibody Species Reactivity: Human

Antibody Positive Control: The observed molecular mass of the processed target protein on SDS-PAGE gels is reported to be around 150-160 kDa.

Antibody Specificity: Very high

Antibody Cross Reactivity: No cross-reactive proteins detected in HeLa cells and in sea star oocytes.

Related Product 1: ASK1-pS1033 blocking peptide

Scientific Background: ASK1 (MAP3K5, MAPKKK5) is a protein-serine/threonine kinase that is a member of the STE group of protein kinases in the STE11 family. This kinase is moderate to highly expressed in most tested human tissues. ASK1 regulates apoptosis through mitochondria-dependent caspase activation. Overexpression of ASK1 induces apoptotic cell death. ASK1 interacts with members of the TRAF family and is activated by TRAF2 in the tumour necrosis factor-alpha (TNF) signalling pathway. ASK1 is activated by phosphorylation at T838 by MAP3K6 and inhibited by phosphorylation at S966 and S1033. It acts as an upstream activator of the MKK>JNK and MKK>p38 MAPK signal transduction cascades through the direct phosphorylation of different MAP kinases, such as MAP2K4 (MKK4), MAP2K3 (MKK3), MAP2K6 (MKK6), and MAP2K7 (MKK7). It is dephosphorylated and activated by PGAM5. ASK1 contains an N-terminal autoinhibitory domain. It is inhibited by phosphorylation at S966 and S1033, and by HIV-1 Nef. ASK1 appears to be a tumour suppressor protein (TSP) as the active form of the protein kinase normally acts to inhibit tumour cell proliferation. 24% of melanoma cell lines were found to contain mutations in the coding-regions of either MAP3K5 or the closely related kinase MAP3K9, often in the kinase catalytic domain.Loss of heterozygosity in MAP3K5 was observed in 85% of melanoma specimens, indicating that the previously observed mutations result in the loss-of-function in the protein. An I780F substitution, the most commonly observed mutant form of the kinase, inhibits phosphotransferase activity.
Figure 1. Epitope mapping of ASK1-pS1033 antibody with similar phosphopeptides on dot blots.


Figure 2. Epitope mapping of ASK1-pS1033 antibody with similar phosphopeptides on dot blots.


Figure 3. Identification of phosphosites related to ASK1-pS1033.