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506123 Anti-p38 MAP Kinase (341-360) Rabbit pAb

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).

Anti-p38 MAP Kinase (341-360) Rabbit pAb MSDS (material safety data sheet) or SDS, CoA and CoQ, dossiers, brochures and other available documents.
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506123
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Overview

Replacement Information

Key Spec Table

Species ReactivityHostAntibody Type
H, M, RRbPolyclonal Antibody

Pricing & Availability

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506123-200UL
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      		 Plastic ampoule 200 ul
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      Description
      OverviewRecognizes 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
      		•
      Catalogue Number506123
      Brand Family Calbiochem®
      Application Data
      Detection of p38 MAP kinase by immunoblotting. Samples: C6 cells (lanes 1 and 2) treated (lane 1) or untreated (lane 2) with anisomycin and NIH/3T3 cells (lanes 3 and 4) treated (lane 3) or untreated (lane 4) with UV. Primary antibody: Anti-p38 MAP Kinase (341-360) Rabbit pAb (Cat. No. 506123) (1:1000). Detection: chemiluminescence.
      References
      ReferencesRaingeaud, J., et al. 1995. J. Biol. Chem. 270, 7420.
      Zervos, A.S., et al. 1995. Proc. Natl. Acad. Sci. USA 92, 10531.
      Han, J., et al. 1994. Science 265, 808.
      Lee, J.C., et al. 1994. Nature 372, 739.
      Rouse, J., et al. 1994. Cell 78, 1027.
      Product Information
      FormLiquid
      FormulationIn 150 mM NaCl, 10 mM HEPES, 50% glycerol, 0.01% BSA, pH 7.5.
      PreservativeNone
      Quality LevelMQ100
      Applications
      Key Applications Flow Cytometry
      Immunoblotting (Western Blotting)
      Paraffin Sections
      Application NotesFlow Cytometry (1:25)
      Immunoblotting (1:1000)
      Paraffin Sections (1:50, heat pretreatment required, see comments)
      Application CommentsPretreat 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.

      Recommended Protocol for Immunoblotting

      Solutions and Reagents
      • Transfer Buffer: 25 mM Tris base, 0.2 M glycine, 20% methanol, pH 8.5.
      • SDS Sample Buffer: 62.5 mM Tris-HCl, pH 6.8, 2% SDS, 10% glycerol, 50 mM DTT, 0.1% bromophenol blue.
      • 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.
      • Blocking Buffer: 1X TBS, 0.1% Tween®-20 detergent with 5% non-fat dry milk.
      • Primary Antibody Dilution Buffer: 1X TBS, 0.1% Tween®-20 detergent with 5% BSA
      • Wash Buffer (TBST): 1X TBS, 0.1% Tween®-20 detergent

      Blotting Membrane
      Nitrocellulose or PVDF membranes may be used.

      Protein Blotting
      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.
      2. Sonicate for 2 s to shear DNA and reduce sample viscosity.
      3. Heat sample to 95-100°C for 5 min. Cool on ice.
      4. Microcentrifuge for 5 min.
      5. Load 20 ml onto SDS-PAGE gel (10 cm x 10 cm).
      6. Electrotransfer to nitrocellulose membrane.

      As controls, we recommend using 15 ml of phosphorylated and nonphosphorylated C-6 glioma cell extracts.

      Membrane Blocking, Gel and Antibody Incubations
      1. After transfer, wash membrane with 25 ml TBS for 5 min at room temperature.
      2. Incubate membrane in 25 ml of Blocking Buffer for 1-3 h at room temperature or overnight at 4°C.
      3. Wash 3 times for 5 min each with 15 ml TBST.
      4. Incubate membrane and primary antibody (at the appropriate dilution) in 10 ml Primary Antibody Dilution Buffer with gentle agitation overnight at 4°C.
      5. Wash 3 times for 5 min each with 15 ml TBST.
      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.
      7. Wash membrane as in step 5.

      Detection of Proteins
      Chemiluminescence.
      Biological Information
      Immunogena synthetic peptide (TYDEYISFVPPPLDQEEMES) corresponding to amino acids 341-360 of human p38 MAP kinase
      ImmunogenHuman
      HostRabbit
      IsotypeIgG
      Species Reactivity
      • Human
      • Mouse
      • Rat
      Antibody TypePolyclonal Antibody
      Physicochemical Information
      Dimensions
      Materials Information
      Toxicological Information
      Safety Information according to GHS
      Safety Information
      Product Usage Statements
      Storage and Shipping Information
      Ship Code Blue Ice Only
      Toxicity Standard Handling
      Storage -20°C
      Avoid freeze/thaw Avoid freeze/thaw
      Do not freeze Ok to freeze
      Special InstructionsFollowing initial thaw, aliquot and freeze (-20°C).
      Packaging Information
      Transport Information
      Supplemental Information
      Specifications
      Global Trade Item Number
      Catalogue Number GTIN
      506123-200UL 04055977199475

      Documentation

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

      Title

      Safety Data Sheet (SDS) 

      Anti-p38 MAP Kinase (341-360) Rabbit pAb Certificates of Analysis

      TitleLot Number
      506123

      References

      Reference overview
      Raingeaud, J., et al. 1995. J. Biol. Chem. 270, 7420.
      Zervos, A.S., et al. 1995. Proc. Natl. Acad. Sci. USA 92, 10531.
      Han, J., et al. 1994. Science 265, 808.
      Lee, J.C., et al. 1994. Nature 372, 739.
      Rouse, J., et al. 1994. Cell 78, 1027.
      Data Sheet

      Note that this data sheet is not lot-specific and is representative of the current specifications for this product. Please consult the vial label and the certificate of analysis for information on specific lots. Also note that shipping conditions may differ from storage conditions.

      Revision17-July-2007 RFH
      ApplicationFlow Cytometry (1:25)
      Immunoblotting (1:1000)
      Paraffin Sections (1:50, heat pretreatment required, see comments)
      Application Data
      Detection of p38 MAP kinase by immunoblotting. Samples: C6 cells (lanes 1 and 2) treated (lane 1) or untreated (lane 2) with anisomycin and NIH/3T3 cells (lanes 3 and 4) treated (lane 3) or untreated (lane 4) with UV. Primary antibody: Anti-p38 MAP Kinase (341-360) Rabbit pAb (Cat. No. 506123) (1:1000). Detection: chemiluminescence.
      DescriptionProtein A and immunoaffinity purified rabbit polyclonal antibody. Recognizes the ~38 kDa p38 MAPK protein.
      Backgroundp38 MAP kinase (MAPK), also called RK and CSBP, is the mammalian homologue of the yeast HOG kinase and participates in a cascade controlling cellular responses to cytokines and stress. Like the SAPK/JNK pathway, p38 MAPK is activated by a variety of cellular stresses including osmotic shock, inflammatory cytokines, lipopolysaccharides (LPS), UV light and growth factors. Both MKK3 and SEK phosphorylate p38 on tyrosine and threonine at the sequence T*GY* resulting in p38 activation. Activated p38 MAPK has been shown to phosphorylate and activate MAPKAP kinase-2 and to phosphorylate the transcription factors ATF-2 and Max.
      HostRabbit
      Immunogen speciesHuman
      Immunogena synthetic peptide (TYDEYISFVPPPLDQEEMES) corresponding to amino acids 341-360 of human p38 MAP kinase
      IsotypeIgG
      Specieshuman, mouse, rat
      FormLiquid
      FormulationIn 150 mM NaCl, 10 mM HEPES, 50% glycerol, 0.01% BSA, pH 7.5.
      PreservativeNone
      CommentsPretreat 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.

      Recommended Protocol for Immunoblotting

      Solutions and Reagents
      • Transfer Buffer: 25 mM Tris base, 0.2 M glycine, 20% methanol, pH 8.5.
      • SDS Sample Buffer: 62.5 mM Tris-HCl, pH 6.8, 2% SDS, 10% glycerol, 50 mM DTT, 0.1% bromophenol blue.
      • 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.
      • Blocking Buffer: 1X TBS, 0.1% Tween®-20 detergent with 5% non-fat dry milk.
      • Primary Antibody Dilution Buffer: 1X TBS, 0.1% Tween®-20 detergent with 5% BSA
      • Wash Buffer (TBST): 1X TBS, 0.1% Tween®-20 detergent

      Blotting Membrane
      Nitrocellulose or PVDF membranes may be used.

      Protein Blotting
      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.
      2. Sonicate for 2 s to shear DNA and reduce sample viscosity.
      3. Heat sample to 95-100°C for 5 min. Cool on ice.
      4. Microcentrifuge for 5 min.
      5. Load 20 ml onto SDS-PAGE gel (10 cm x 10 cm).
      6. Electrotransfer to nitrocellulose membrane.

      As controls, we recommend using 15 ml of phosphorylated and nonphosphorylated C-6 glioma cell extracts.

      Membrane Blocking, Gel and Antibody Incubations
      1. After transfer, wash membrane with 25 ml TBS for 5 min at room temperature.
      2. Incubate membrane in 25 ml of Blocking Buffer for 1-3 h at room temperature or overnight at 4°C.
      3. Wash 3 times for 5 min each with 15 ml TBST.
      4. Incubate membrane and primary antibody (at the appropriate dilution) in 10 ml Primary Antibody Dilution Buffer with gentle agitation overnight at 4°C.
      5. Wash 3 times for 5 min each with 15 ml TBST.
      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.
      7. Wash membrane as in step 5.

      Detection of Proteins
      Chemiluminescence.
      Storage Avoid freeze/thaw
      -20°C
      Do Not Freeze Ok to freeze
      Special InstructionsFollowing initial thaw, aliquot and freeze (-20°C).
      Toxicity Standard Handling
      ReferencesRaingeaud, J., et al. 1995. J. Biol. Chem. 270, 7420.
      Zervos, A.S., et al. 1995. Proc. Natl. Acad. Sci. USA 92, 10531.
      Han, J., et al. 1994. Science 265, 808.
      Lee, J.C., et al. 1994. Nature 372, 739.
      Rouse, J., et al. 1994. Cell 78, 1027.