Anti-p38 MAP Kinase (341-360) Rabbit pAb

Application

Flow Cytometry (1:25)r>Immunoblotting (1:1000)r>Paraffin Sections (1:50, heat pretreatment required, see comments)

General description

Recognizes the ~38 kDa p38 MAPK protein.Antibody Target Gene Symbol: MAPK14 Target Synonym: CRK1, CSBP, CSBP1, CSBP2, CSPB1, EXIP, Hog, MAPK p38, MGC102436, MGC105413, MXI2, P38, P38 KINASE, P38 Map Kinase, p38 Mapk alpha, P38-ALPHA, p38-RK, p38/Hog1, p38/Mpk2, P38/RK, p38a, p38Hog, p38MAPK, PRKM14, PRKM15, RK, SAPK2A Entrez Gene Name: mitogen-activated protein kinase 14 Hu Entrez ID: 1432 Mu Entrez ID: 26416 Rat Entrez ID: 81649

Protein A and immunoaffinity purified rabbit polyclonal antibody. Recognizes the ~38 kDa p38 MAPK protein.

This Anti-p38 MAP Kinase (341-360) Rabbit pAb is validated for use in Flow Cytometry, Immunoblotting, Paraffin Sections for the detection of p38 MAP Kinase (341-360).

Immunogen

a synthetic peptide (TYDEYISFVPPPLDQEEMES) corresponding to amino acids 341-360 of human p38 MAP kinase

Human

Legal Information

TWEEN is a registered trademark of Croda International PLC

CALBIOCHEM is a registered trademark of Merck KGaA, Darmstadt, Germany

Other Notes

Pretreat paraffin sections by heating tissue in 10 mM citrate buffer, pH 6.0 for 1 min at high power followed by 9 min at medium power; keep the slides fully immersed and maintain the temperature at or just below boiling; cool the slides for 20 min at room temperature prior to staining. Variables associated with assay conditions will dictate the proper working dilution.r>r>Recommended Protocol for Immunoblottingr>r>Solutions and Reagentsr> Transfer Buffer: 25 mM Tris base, 0.2 M glycine, 20% methanol, pH 8.5.r> SDS Sample Buffer: 62.5 mM Tris-HCl, pH 6.8, 2% SDS, 10% glycerol, 50 mM DTT, 0.1% bromophenol blue.r> 10X TBS (Tris-buffered saline): To prepare 1 liter, 24.2 g Tris base, 80 g NaCl, adjust pH to 7.6 with HCl. Dilute 1:10 for use.r> Blocking Buffer: 1X TBS, 0.1% Tween^®-20 detergent with 5% non-fat dry milk.r> Primary Antibody Dilution Buffer: 1X TBS, 0.1% Tween™-20 detergent with 5% BSAr> Wash Buffer (TBST): 1X TBS, 0.1% Tween™-20 detergentr>r>Blotting Membraner>Nitrocellulose or PVDF membranes may be used.r>r>Protein Blottingr>1. Lyse cells by adding 100 ml SDS Sample Buffer and immediately scrape the cells off the plate and transfer the extract to a microfuge tube. Keep on ice.r>2. Sonicate for 2 s to shear DNA and reduce sample viscosity.r>3. Heat sample to 95-100°C for 5 min. Cool on ice.r>4. Microcentrifuge for 5 min.r>5. Load 20 ml onto SDS-PAGE gel (10 cm x 10 cm).r>6. Electrotransfer to nitrocellulose membrane.r>r>As controls, we recommend using 15 ml of phosphorylated and nonphosphorylated C-6 glioma cell extracts.r>r>Membrane Blocking, Gel and Antibody Incubationsr>1. After transfer, wash membrane with 25 ml TBS for 5 min at room temperature.r>2. Incubate membrane in 25 ml of Blocking Buffer for 1-3 h at room temperature or overnight at 4°C.r>3. Wash 3 times for 5 min each with 15 ml TBST.r>4. Incubate membrane and primary antibody (at the appropriate dilution) in 10 ml Primary Antibody Dilution Buffer with gentle agitation overnight at 4°C.r>5. Wash 3 times for 5 min each with 15 ml TBST.r>6. Incubate membrane with conjugated secondary antibody at the appropriate dilution in 10 ml Blocking Buffer with gentle agitation for 1 h at room temperature.r>7. Wash membrane as in step 5.r>r>Detection of Proteinsr>Chemiluminescence.

Raingeaud, J., et al. 1995. J. Biol. Chem.270, 7420.Zervos, A.S., et al. 1995. Proc. Natl. Acad. Sci. USA92, 10531.Han, J., et al. 1994. Science265, 808.Lee, J.C., et al. 1994. Nature372, 739.Rouse, J., et al. 1994. Cell78, 1027.

Packaging

200 µL in Plastic ampoule

Physical form

In 150 mM NaCl, 10 mM HEPES, 50% glycerol, 0.01% BSA, pH 7.5.

Reconstitution

Following initial thaw, aliquot and freeze (-20°C).

Warning

Toxicity: Standard Handling (A)