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559304 Anti-SAPK/JNK Rabbit pAb

This Anti-SAPK/JNK Rabbit pAb is validated for use in Immunoblotting, Immunocytochemistry for the detection of SAPK/JNK.

Anti-SAPK/JNK Rabbit pAb : FDS (Fiches de données de sécurité), certificats d’analyse (CoA) et de qualité (CoQ), dossiers, brochures et autres documents disponibles.
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559304
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Replacement Information

Tableau de caractéristiques principal

Species ReactivityHostAntibody Type
H, M, RRbPolyclonal Antibody

Prix & Disponibilité

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559304-200UL
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      Description
      OverviewRecognizes the ~54 kDa SAPK/JNK protein in uv-treated HEK293 cells.
      Catalogue Number559304
      Brand Family Calbiochem®
      Application Data
      Detection of human SAPK/JNK by immunoblotting. Sample: Whole cell lysate from HEK 293 cells and SK-N-MC cells treated with UV (40 J/m2) for the indicated times. Primary antibody: Anti-SAPK/JNK Rabbit pAb (Cat. No. 559304) (1:1000). Detection: chemiluminescence.
      References
      ReferencesGupta, S., et al. 1996. EMBO J. 15(11), 2760.
      Coso, O.A., et al. 1995. Cell 81, 1137.
      Derijard, B., et al. 1994. Cell 76, 1025.
      Kyriakis, J.M., et al. 1994. Nature 369, 156.
      Hibi, M., et al. 1993. Genes Dev. 7, 2135.
      Kyriakis, J.M. and Avruch, J. 1990. J. Biol. Chem. 265, 17355.
      Product Information
      FormLiquid
      FormulationIn 150 mM NaCl, 10 mM HEPES, 50% glycerol, 0.01% BSA, pH 7.5.
      Positive controlUV treated HEK293 cells
      PreservativeNone
      Quality LevelMQ100
      Applications
      Key Applications Immunoblotting (Western Blotting)
      Immunocytochemistry
      Application NotesImmunoblotting (1:1000)
      Immunocytochemistry (1:200)
      Application CommentsRecognizes SAPK/JNK regardless of the phosphorylation state. 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% bromphenol 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
      A general protocol for sample preparation using 2x106 293 cells per well in a 6-well plate is as follows:

      1. Aspirate media. Treat cells by adding fresh media containing regulator for desired time.
      2. Aspirate media from cultures; wash cells with PBS; aspirate.
      3. Lyse cells by adding 100 µl of SDS Sample Buffer and immediately scrape the cells off the plate and transfer the extract to a microfuge tube. Keep on ice.
      4. Sonicate for 2 s to shear DNA and reduce sample viscosity.
      5. Heat sample to 95-100°C for 5 min. Cool on ice.
      6. Microcentrifuge for 5 min.
      7. Load 20 µl onto SDS-PAGE gel (10 cm x 10 cm).
      8. Electrotransfer to nitrocellulose membrane.

      As controls, we recommend using 20 µl lysate from UV treated HEK293 cell.

      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 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 full-length, recombinant, human p54 SAPK/JNK2 fusion protein
      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
      Référence GTIN
      559304-200UL 04055977192421

      Documentation

      Anti-SAPK/JNK Rabbit pAb FDS

      Titre

      Fiche de données de sécurité des matériaux (FDS) 

      Anti-SAPK/JNK Rabbit pAb Certificats d'analyse

      TitreNuméro de lot
      559304

      Références bibliographiques

      Aperçu de la référence bibliographique
      Gupta, S., et al. 1996. EMBO J. 15(11), 2760.
      Coso, O.A., et al. 1995. Cell 81, 1137.
      Derijard, B., et al. 1994. Cell 76, 1025.
      Kyriakis, J.M., et al. 1994. Nature 369, 156.
      Hibi, M., et al. 1993. Genes Dev. 7, 2135.
      Kyriakis, J.M. and Avruch, J. 1990. J. Biol. Chem. 265, 17355.
      Fiche technique

      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.

      Revision15-August-2007 RFH
      ApplicationImmunoblotting (1:1000)
      Immunocytochemistry (1:200)
      Application Data
      Detection of human SAPK/JNK by immunoblotting. Sample: Whole cell lysate from HEK 293 cells and SK-N-MC cells treated with UV (40 J/m2) for the indicated times. Primary antibody: Anti-SAPK/JNK Rabbit pAb (Cat. No. 559304) (1:1000). Detection: chemiluminescence.
      DescriptionProtein A and immunoaffinity purified rabbit polyclonal antibody. Recognizes the ~54 kDa SAPK/JNK protein.
      BackgroundThe Stress-activated protein kinases (SAPK), also referred to as Jun N-terminal kinases (JNK), function in a protein kinase cascade transducing cellular stress signals. SAPK/JNKs are activated by a highly diverse group of extracellular signals, including UV light, inflammatory cytokines and a wide variety of cellular stresses. Activation occurs via phosphorylation at Thr183 and Tyr185 by the dual specificity enzyme SEK/MKK4 and as yet unidentified kinases. Since dual phosphorylation of SAPK/JNK at Thr183/Tyr185 is essential for kinase activity, phosphorylation at this site is an excellent marker of SAPK/JNK activity. Activated SAPK in turn phosphorylates and activates several transcription factors, including c-jun at Ser63/73 and ATF-2 at Ser69/71.
      HostRabbit
      Immunogen speciesHuman
      Immunogena full-length, recombinant, human p54 SAPK/JNK2 fusion protein
      IsotypeIgG
      Specieshuman, mouse, rat
      Positive controlUV treated HEK293 cells
      FormLiquid
      FormulationIn 150 mM NaCl, 10 mM HEPES, 50% glycerol, 0.01% BSA, pH 7.5.
      PreservativeNone
      CommentsRecognizes SAPK/JNK regardless of the phosphorylation state. 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% bromphenol 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
      A general protocol for sample preparation using 2x106 293 cells per well in a 6-well plate is as follows:

      1. Aspirate media. Treat cells by adding fresh media containing regulator for desired time.
      2. Aspirate media from cultures; wash cells with PBS; aspirate.
      3. Lyse cells by adding 100 µl of SDS Sample Buffer and immediately scrape the cells off the plate and transfer the extract to a microfuge tube. Keep on ice.
      4. Sonicate for 2 s to shear DNA and reduce sample viscosity.
      5. Heat sample to 95-100°C for 5 min. Cool on ice.
      6. Microcentrifuge for 5 min.
      7. Load 20 µl onto SDS-PAGE gel (10 cm x 10 cm).
      8. Electrotransfer to nitrocellulose membrane.

      As controls, we recommend using 20 µl lysate from UV treated HEK293 cell.

      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 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
      ReferencesGupta, S., et al. 1996. EMBO J. 15(11), 2760.
      Coso, O.A., et al. 1995. Cell 81, 1137.
      Derijard, B., et al. 1994. Cell 76, 1025.
      Kyriakis, J.M., et al. 1994. Nature 369, 156.
      Hibi, M., et al. 1993. Genes Dev. 7, 2135.
      Kyriakis, J.M. and Avruch, J. 1990. J. Biol. Chem. 265, 17355.