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NA57 Anti-DNA-PK (Ab-2) Mouse mAb (18-2)

Overview

Replacement Information

Key Spec Table

Species ReactivityHostAntibody Type
HMMonoclonal Antibody

Products

Catalogue NumberPackaging Qty/Pack
NA57-100UG Plastic ampoule 100 μg
Description
OverviewRecognizes the ~470 kDa DNA-PK protein in A431, HeLa, and HL60 cells, normal tonsil, normal colon, and breast carcinoma tissue.
Catalogue NumberNA57
Brand Family Calbiochem®
SynonymsAnti-DNA Dependent Protein Kinase
Application Data
Detection of human DNA-PK by immunohistochemistry. Sample: Human tonsil tissue fixed in formalin and embedded in paraffin. Primary antibody: Anti-DNA-PK (Ab-2) Mouse mAb (18-2) (Cat. No. NA57) (1:100). Detection: fluorescence.

Detection of human DNA-PK by immunoblotting. Sample: Human DNA-PK. Primary antibody: Anti-DNA-PK (Ab-2) Mouse mAb (18-2) (Cat. No. NA57) (1 µg/ml). Detection: chemiluminescence.
References
References<→EMD:if(output-is PDS)→> Kharbanda, S., et al. 1997. Nature 386, 732.
McConnell, K.R., et al. 1997 J. Immunol. 158, 2083-2089.
Peterson, S.R., et al. 1997. J. Biol. Chem. 272, 10227.
Anderson, C.W. and Carter, T.H. 1996. Curr. Top. Microbiol. Immunol. 217, 91.
Blunt, T., et al. 1996. Proc. Natl. Acad. Sci. USA 93, 10285.
Carpenter, C.L. and Cantley, L.C. 1996. Curr. Opin. Cell. Biol. 8, 153.
Chan, D.W. and Lees-Miller, S.P. 1996. J. Biol. Chem. 271, 8936.
Connelly, M.A., et al. 1996. Gene 175, 271.
Danska, J.S., et al. 1996. Mol. Cell. Biol. 16, 5507.
Han, Z., et al. 1996. J. Biol. Chem. 271, 25035.
Lees-Miller, S.P. 1996. Biochem. Cell. Biol. 74, 503.
Song, Q., et al. 1996. EMBO J. 15, 3238.
Teraoka, H., et al. 1996. FEBS Lett. 393, 1.
Blunt, T., et al. 1995. Cell 80, 813.
Hartley, K.O., et al. 1995. Cell 82, 849.
Kirchgessner, C.U., et al. 1995. Science 267, 1178.
Lees-Miller, S.P., et al. 1995. Science 267, 1183.
Poltoratsky, V.P., et al. 1995. J. Immunol. 155, 4529.
Anderson, C.W. 1993. Trends Biochem. Sci. 18, 433.
Gottleib, T.M. and Jackson, S.P. 1993. Cell 72, 131. <→EMD:endif()→>
Product Information
FormLiquid
FormulationIn 10 mM PBS, 0.2% BSA, pH 7.4.
Positive controlA-431, HeLa, or HL-60 cells or normal tonsil, normal colon, or breast carcinoma tissue
Preservative≤0.1% sodium azide
Quality LevelMQ100
Applications
Application ReferencesOriginal Clone Carter, T., et al. 1990. Mol. Cell. Biol. 10, 6460.
Key Applications Frozen Sections
Immunoblotting (Western Blotting)
Immunofluorescence
Immunoprecipitation
Neutralization Studies
Paraffin Sections
Application NotesFrozen Sections (1:50-1:100)
Immunoblotting (1:250-1:500)
Immunofluorescence (1:50-1:100)
Immunoprecipitation (10 µl/mg protein lysate)
Neutralization Studies (see comments)
Paraffin Sections (1:50-1:100, heat/pressure cooker pre-treatment required)
Application CommentsThis antibody will inhibit (approximately 50%) the kinase activity of DNA-PKcs. However, preincubation of the enzyme with DNA protects DNA-PKcs against inactivation by Clone 18-2. Antibody should be titrated for optimal results in individual systems.
Biological Information
ImmunogenDNA-PK purified from HeLa cells
ImmunogenHuman
Epitopewithin amino acids 1-2713 of DNA-PK
Clone18-2
HostMouse
IsotypeIgG₁
Species Reactivity
  • Human
Antibody TypeMonoclonal Antibody
Concentration Label Please refer to vial label for lot-specific concentration
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 +2°C to +8°C
Do not freeze Ok to freeze
Special InstructionsFor long-term storage, aliquot and freeze (-20°C). Avoid freeze/thaw cycles.
Packaging Information
Transport Information
Supplemental Information
Specifications
Global Trade Item Number
Catalogue Number GTIN
NA57-100UG 04055977227598

Documentation

Anti-DNA-PK (Ab-2) Mouse mAb (18-2) SDS

Title

Safety Data Sheet (SDS) 

Anti-DNA-PK (Ab-2) Mouse mAb (18-2) Certificates of Analysis

TitleLot Number
NA57

References

Reference overview
<→EMD:if(output-is PDS)→> Kharbanda, S., et al. 1997. Nature 386, 732.
McConnell, K.R., et al. 1997 J. Immunol. 158, 2083-2089.
Peterson, S.R., et al. 1997. J. Biol. Chem. 272, 10227.
Anderson, C.W. and Carter, T.H. 1996. Curr. Top. Microbiol. Immunol. 217, 91.
Blunt, T., et al. 1996. Proc. Natl. Acad. Sci. USA 93, 10285.
Carpenter, C.L. and Cantley, L.C. 1996. Curr. Opin. Cell. Biol. 8, 153.
Chan, D.W. and Lees-Miller, S.P. 1996. J. Biol. Chem. 271, 8936.
Connelly, M.A., et al. 1996. Gene 175, 271.
Danska, J.S., et al. 1996. Mol. Cell. Biol. 16, 5507.
Han, Z., et al. 1996. J. Biol. Chem. 271, 25035.
Lees-Miller, S.P. 1996. Biochem. Cell. Biol. 74, 503.
Song, Q., et al. 1996. EMBO J. 15, 3238.
Teraoka, H., et al. 1996. FEBS Lett. 393, 1.
Blunt, T., et al. 1995. Cell 80, 813.
Hartley, K.O., et al. 1995. Cell 82, 849.
Kirchgessner, C.U., et al. 1995. Science 267, 1178.
Lees-Miller, S.P., et al. 1995. Science 267, 1183.
Poltoratsky, V.P., et al. 1995. J. Immunol. 155, 4529.
Anderson, C.W. 1993. Trends Biochem. Sci. 18, 433.
Gottleib, T.M. and Jackson, S.P. 1993. Cell 72, 131. <→EMD:endif()→>
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.

Revision05-September-2007 RFH
SynonymsAnti-DNA Dependent Protein Kinase
ApplicationFrozen Sections (1:50-1:100)
Immunoblotting (1:250-1:500)
Immunofluorescence (1:50-1:100)
Immunoprecipitation (10 µl/mg protein lysate)
Neutralization Studies (see comments)
Paraffin Sections (1:50-1:100, heat/pressure cooker pre-treatment required)
Application Data
Detection of human DNA-PK by immunohistochemistry. Sample: Human tonsil tissue fixed in formalin and embedded in paraffin. Primary antibody: Anti-DNA-PK (Ab-2) Mouse mAb (18-2) (Cat. No. NA57) (1:100). Detection: fluorescence.

Detection of human DNA-PK by immunoblotting. Sample: Human DNA-PK. Primary antibody: Anti-DNA-PK (Ab-2) Mouse mAb (18-2) (Cat. No. NA57) (1 µg/ml). Detection: chemiluminescence.
DescriptionPurified mouse monoclonal antibody generated by immunizing BALB/c mice with the specified immunogen and fusing the splenocytes with mouse myeloma SP2/0 cells (see application references). Recognizes the ~460 kDa DNA-PK protein.
BackgroundDNA Dependent Protein Kinase, DNA-PK, is a nuclear serine/threonine protein kinase composed of two subunits, a ~470 kDa catalytic subunit, DNA-PKcs, and the Ku autoantigen, the heterodimeric DNA binding protein p70/p80. DNA-PK has been shown to phosphorylate a number of key proteins in vitro including p53, Sp1, myc, RPA, and RNA Pol I. DNA-PK activity requires association of DNA-PKcs with Ku, DNA binding, and the presence of DNA ends. Mutations in DNA-PKcs contribute to murine SCID phenotypes which leads to inefficient V(D)J recombination in developing T & B lymphocytes. Loss of DNA-PK activity in fibroblasts or lymphocytes also confers on cells hypersensitivity to killing by ionizing radiation. These two phenotypes implicate DNA-PK in the repair of DNA double strand breaks in all cells and V(D)J recombination in lymphocytes. The 4127 amino acid DNA-PKcs protein shares a high degree of similarity with the ATM family of kinases, including ATM, ATR, and FRAP in humans and MEC1 and TEL1 in yeast. The catalytic domain of DNA-PKcs also shares homology with the PI3K family of lipid kinases and DNA-PK is inhibitable by wortmanin, a lipid kinase inhibitor. While the exact role for DNA-PK in cell cycle regulation is unclear, the protein may be important for suppression of apoptosis since DNA-PKcs is targeted for proteolysis during apoptosis, which leads to loss of catalytic activity. DNA-PK can also be inactivated through phosphorylation, possibly by c-abl and by autophosphorylation. DNA-PKcs is expressed in all cells and a DNA-PK activity is observed in all higher eukaryotes examined. The protein level of DNA-PKcs in human cells is very high, 50 to 100x that observed in murine cells.
HostMouse
Immunogen speciesHuman
ImmunogenDNA-PK purified from HeLa cells
Epitopewithin amino acids 1-2713 of DNA-PK
Clone18-2
IsotypeIgG₁
Specieshuman
Positive controlA-431, HeLa, or HL-60 cells or normal tonsil, normal colon, or breast carcinoma tissue
FormLiquid
FormulationIn 10 mM PBS, 0.2% BSA, pH 7.4.
Concentration Label Please refer to vial label for lot-specific concentration
Preservative≤0.1% sodium azide
CommentsThis antibody will inhibit (approximately 50%) the kinase activity of DNA-PKcs. However, preincubation of the enzyme with DNA protects DNA-PKcs against inactivation by Clone 18-2. Antibody should be titrated for optimal results in individual systems.
Storage +2°C to +8°C
Do Not Freeze Ok to freeze
Special InstructionsFor long-term storage, aliquot and freeze (-20°C). Avoid freeze/thaw cycles.
Toxicity Standard Handling
References<→EMD:if(output-is PDS)→> Kharbanda, S., et al. 1997. Nature 386, 732.
McConnell, K.R., et al. 1997 J. Immunol. 158, 2083-2089.
Peterson, S.R., et al. 1997. J. Biol. Chem. 272, 10227.
Anderson, C.W. and Carter, T.H. 1996. Curr. Top. Microbiol. Immunol. 217, 91.
Blunt, T., et al. 1996. Proc. Natl. Acad. Sci. USA 93, 10285.
Carpenter, C.L. and Cantley, L.C. 1996. Curr. Opin. Cell. Biol. 8, 153.
Chan, D.W. and Lees-Miller, S.P. 1996. J. Biol. Chem. 271, 8936.
Connelly, M.A., et al. 1996. Gene 175, 271.
Danska, J.S., et al. 1996. Mol. Cell. Biol. 16, 5507.
Han, Z., et al. 1996. J. Biol. Chem. 271, 25035.
Lees-Miller, S.P. 1996. Biochem. Cell. Biol. 74, 503.
Song, Q., et al. 1996. EMBO J. 15, 3238.
Teraoka, H., et al. 1996. FEBS Lett. 393, 1.
Blunt, T., et al. 1995. Cell 80, 813.
Hartley, K.O., et al. 1995. Cell 82, 849.
Kirchgessner, C.U., et al. 1995. Science 267, 1178.
Lees-Miller, S.P., et al. 1995. Science 267, 1183.
Poltoratsky, V.P., et al. 1995. J. Immunol. 155, 4529.
Anderson, C.W. 1993. Trends Biochem. Sci. 18, 433.
Gottleib, T.M. and Jackson, S.P. 1993. Cell 72, 131. <→EMD:endif()→>
Application referencesOriginal Clone Carter, T., et al. 1990. Mol. Cell. Biol. 10, 6460.