Mitogen-Activated Protein Kinase-Activated Protein Kinase 2 (MAPKAPK2) antibody

Antigen: Mitogen-Activated Protein Kinase-Activated Protein Kinase 2 (MAPKAPK2)

Synonyms: MK2, Rps6kc1, AA960234, mapkapk2, wu:fb30e10, wu:fc56c10, wu:fi03a01, MGC78852, MAPKAPK2

Clonality: Polyclonal

Host: Rabbit

Reactivity: Human, Cow (Bovine), Mouse (Murine), Rat (Rattus)

Application: ELISA, Western Blotting (WB), Immunoprecipitation (IP)

Catalog no.: ABIN103960

Additional Information

Alternative name: MAPKAP Kinase 2

Immunogen: This antibody was affinity purified from whole rabbit serum prepared by repeated immunizationswith a synthetic peptide corresponding to aa 310-325 of rabbit MAPKAP Kinase 2 conjugated to KLH using maleimide. A terminal cysteine residue was added to facilitate coupling. Peptide Sequence: M-T-S-A-L-A-T-M-R-V-D-Y-E-Q-I-K

Format: Liquid (sterile filtered)

Description: MAPKAP kinase-2 (or MK2) is anacronym for mitogen-activated protein kinase-activated proteinkinase-2. This enzyme is phosphorylated and activated by Erksand p38 MAPK in vitro. While multiple residues of MK2 arephosphorylated in vivo in response to stress, only four of theresidues Thr25, Thr222, Ser272 and Thr334 are phosphorylatedby p38 in an in vitro kinase assay. MK2 is serine-threonineprotein kinase that contains a proline rich sequence and twoputative SH3-binding sites. MK2 phosphorylates downstreamcomponents on a consensus Pro-X-Ser/Thr-Pro motif. The best-characterized MK2 substrate is hsp27. Although unknown, thephysiological function of MK2 may be to increase stressresistance. MK2 is an essential component in the inflammatoryresponse that regulates biosynthesis of TNF-alpha at a post- transcriptional level and MK2 is rapidly phosphorylated andactivated in response to cytokines, stress and chemotacticfactors.

Specificity: This affinity-purified antibody is directed against rabbit MAPKAP Kinase 2 and is useful in determiningits presence in various assays. This polyclonal anti-MAPKAP Kinase 2 antibody recognizes native and over-expressed proteinfound in various tissues and extracts. This sequence is conserved 100% in human, mouse, rabbit, rat, hamster, chicken,Xenopus and zebrafish. Cross reactivity with MAPKAP Kinase 2 from other sources has not been tested.

Application Details

Application Notes: Recommended Dilutions: This product was assayed by immunoblot and was found to be reactive to 20 u g of crude protein extract prepared from HeLa cells. The gel was probed with antibodies-online's Affinity Purified anti-MAPKAP Kinase2 used at a dilution of 1:2000. Use Peroxidase Conjugated Affinity Purified Anti-RABBIT IgG (H&L) (GOAT) (Min X Bv Ch GtGP Hs Hu Ms Rt & Sh Serum Proteins) (code ABIN102010) as a secondary antibody for detection. The antibody detects asingle 44 kDa protein in crude extracts prepared either from unstimulated (code w09-000-364) or TNF alpha stimulated (code w09- 000-367) human HeLa cell lysates. A 1:15,000 dilution is recommended for ELISA. The researcher should determine optimaltiters for other applications. Note: This antibody is suitable for westernblotting, ELISA, immunoprecipitation and kinase assays.Although not tested, this antibody is likely functional inimmunohistochemistry and other immunological methods.Affinity purified anti-MAPKAP Kinase 2 is specific for MAPKAP Kinase 2 and will not effectively immunoblot other isoforms of the kinase (i.e. MAPKAP K1 or K3) from cell lines and/or tissues.Use a HeLa cell lysate as a positive control. MK2 assays are often used and are easier and have less background than p38kinase assays. Immunoprecipitation of MK2 using antibodies is used for this purpose.

Concentration: 0.9 mg/ml (by UV absorbance at 280 nm)

Buffer: Stabilizer: None. Preservative: 0.01% (w/v) Sodium Azide. Buffer: 0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2.

Storage: Store vial at -20° C. For extended storage aliquot contents and freeze at -20° C or below. Avoid cyclesof freezing and thawing. Dilute only prior to immediate use. Expiration date is one (1) year from date of restoration.

Research Area: Signaling, Cytoskeleton, Microfilaments, Inflammation, Kinases/Phosphatases

Restrictions: For Research Use only

Images

Affinity purified anti-MAPKAP Kinase 2 polyclonal antibody detects MK2 in unstimulated human HeLa whole cell lysate by western blot. Polyclonal rabbitanti- MAPKAP Kinase 2 used at a 1:2000 dilution to detect 20 ug of whole cell lysate containing MK2. This antibody detects a single 44 kDa protein as indicated in crude extracts prepared from either unstimulated or TNFalpha stimulated human HeLa cell lysates. A 4-20% gradient gel was used to separate the protein by SDS-PAGE. The protein was transferred to nitrocellulose using standard methods. After blocking the membrane was probed with the primary antibody for 1 h at room temperature followed by washes and reaction with a 1:5,000 dilution of IRDye