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OP109 Anti-p73 (Ab-2) Mouse mAb (ER-15)

OP109
  
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      Overview

      Replacement Information

      Key Spec Table

      Species ReactivityHostAntibody Type
      H, Mk M Monoclonal Antibody
      Description
      Overview

      This product has been discontinued.



      Recognizes the ~80 kDa α and the ~68 kDa β isoforms of p73 in HEK293, LOS, and IM9 cells. Will immunoprecipitate p73α/p73β complexes.

      Catalogue NumberOP109
      Brand Family Calbiochem®
      References
      ReferencesDi Como, C.J., et al. 1999. Mol. Cell Biol. 19, 1438.
      De Laurenzi, V., et al. 1998. J. Exp. Med. 188, 1763.
      Dobbelstein, M. and J. Roth. 1998. J. Gen. Virol. 79, 3079.
      Higashino, F., et al. 1998. Proc. Natl. Acad. Sci. USA 95, 15683.
      Marin, M.C., et al. 1998. Mol. Cell Biol. 18, 6316.
      Prabhu, N.S., et al. 1998. Int. J. Oncol. 13, 5.
      Roth, J., et al. 1998. J. Virol. 72, 8510.
      Zhu, J., et al. 1998. Cancer Res 58, 5061.
      Jost, C.A., et al. 1997. Nature 389, 191.
      Kaghad, M., et al. 1997. Cell 90, 809.
      Product Information
      FormLiquid
      FormulationIn 0.05 M sodium phosphate buffer, 0.2% gelatin.
      Negative controlSK-N-AS cells
      Positive controlHEK293, COS, or IM9 cells
      Preservative≤0.1% sodium azide
      Applications
      Application ReferencesOriginal Clone Marin, M.C., et al. 1998. Mol. Cell Biol. 18, 6316.
      Key Applications Gel Shift
      Immunoblotting (Western Blotting)
      Immunoprecipitation
      Application NotesGel Shift (see comments)
      Immunoblotting (2-4 µg/ml)
      Immunoprecipitation (2-4 µg)
      Application CommentsTwo p73 isoforms are expressed in most cells. p73α migrates as an ~80 kDa protein and p73β migrates as a ~68 kDa protein. This antibody detects both p73α and p73β in immunoblots and will immunoprecipitate p73α, p73β, and p73α/p73β complexes. This antibody will also supershift p73α/DNA and p73β/DNA complexes. For gel shift, use Cat. No. OP109L and resuspend in 100 µl buffer. For blotting, use 150 to 300 µg total cell lysate per lane. Antibody should be titrated for optimal results in individual systems.

      1. Using fresh lysates, display 150-300 µg of total cellular protein on 10% PAGE/SDS. Transfer to nitrocellulose.
      2. Block nitrocellulose for 30 min at RT in TBS/T (10 mM Tris-HCl, pH 8.0, 150 mM NaCl, 0.2% Tween®-20 detergent) buffer containing 5% non-fat dry milk and 1% normal goat serum.
      3. Incubate blot with primary antibody in TBS/T (10 mM Tris-HCl, pH 8.0, 150 mM NaCl, 0.2% Tween®-20 detergent) for 2 h at RT or overnight at 4°C.
      4. Wash blot 3 times for 10 min. each with TBS/T.
      5. Incubate blot with goat anti-mouse IgG conjugated to HRP at 50 ng/mL in TBS/T for 30 min at RT.
      6. Wash blot 3 times for 10 min. each with TBS/T.
      7. Detect bound antibodies using chemiluminescence.
      8. Multiple bands may appear in all lanes due to non-specific cross-reacting material. However, bands corresponding to p73α and p73β should be seen in 293 but not in SK-N-AS lysates.
      Biological Information
      Immunogenrecombinant, human p73α protein
      ImmunogenHuman
      Epitopewithin amino acids 380-495 of human p73α
      CloneER-15
      HostMouse
      IsotypeIgG₁
      Species Reactivity
      • Human
      • Monkey
      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 Yes
      Packaging Information
      Transport Information
      Supplemental Information
      Specifications

      Documentation

      Anti-p73 (Ab-2) Mouse mAb (ER-15) MSDS

      Title

      Safety Data Sheet (SDS) 

      Anti-p73 (Ab-2) Mouse mAb (ER-15) Certificates of Analysis

      TitleLot Number
      OP109

      References

      Reference overview
      Di Como, C.J., et al. 1999. Mol. Cell Biol. 19, 1438.
      De Laurenzi, V., et al. 1998. J. Exp. Med. 188, 1763.
      Dobbelstein, M. and J. Roth. 1998. J. Gen. Virol. 79, 3079.
      Higashino, F., et al. 1998. Proc. Natl. Acad. Sci. USA 95, 15683.
      Marin, M.C., et al. 1998. Mol. Cell Biol. 18, 6316.
      Prabhu, N.S., et al. 1998. Int. J. Oncol. 13, 5.
      Roth, J., et al. 1998. J. Virol. 72, 8510.
      Zhu, J., et al. 1998. Cancer Res 58, 5061.
      Jost, C.A., et al. 1997. Nature 389, 191.
      Kaghad, M., et al. 1997. Cell 90, 809.

      Brochure

      Title
      Caspases and other Apoptosis Related Tools Brochure
      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.

      Revision03-December-2007 JSW
      ApplicationGel Shift (see comments)
      Immunoblotting (2-4 µg/ml)
      Immunoprecipitation (2-4 µg)
      DescriptionPurified mouse monoclonal antibody generated by immunizing RBF/DnJ mice with the specified immunogen and fusing splenocytes to NS1 mouse myeloma cells (see application references). Recognizes the ~80 kDa α and the ~68 kDa β isoforms of p73.
      BackgroundThe p73 protein shares considerable structural and functional similarity to p53. The p73 gene encodes 4 polypeptides through alternative splicing of exons 11-13 with p73α and p73β expressed in all tissues while expression of p73γ and p73δ may be more restricted. All isoforms show significant sequence identity to the transactivation, DNA binding, and oligomerization domains of p53, and many of the critical amino acid residues in p53 essential for DNA binding are conserved in p73. Not surprisingly, p73 binds to a p53 consensus sequence and transactivates many, but not all, p53-responsive genes. Overproduction of p73 leads to growth arrest and induction of apoptosis in a p53-independent manner. However, p73 does not transactivate all p53-responsive genes and the level of transactivation is often lower than that by p53; thus, the mechanisms of growth arrest and induction of apoptosis by p73 are at least partially distinct from p53. p73 does not appear to be targeted by many of the viral oncoproteins such as E1B 55K, SV40 T antigen, and HPV E6 that bind to and inactivate p53, however both p53 and p73 do bind the Adenovirus E4orf6 protein, suggesting that p73 may be independently inactivated by viral oncoproteins. All p73 isoforms bind one another and p73α and p73β appear to bind p53 as well. Interestingly, mutant p53 binds p73α and inhibits transcriptional activation by p73 indicating that mutational inactivation of p53 may serve to simultaneously inactivate p73. One key difference between p53 and p73 is that, unlike p53, p73 does not appear to be activated in response to DNA damage.
      HostMouse
      Immunogen speciesHuman
      Immunogenrecombinant, human p73α protein
      Epitopewithin amino acids 380-495 of human p73α
      CloneER-15
      IsotypeIgG₁
      Specieshuman, monkey
      Positive controlHEK293, COS, or IM9 cells
      Negative controlSK-N-AS cells
      FormLiquid
      FormulationIn 0.05 M sodium phosphate buffer, 0.2% gelatin.
      Concentration Label Please refer to vial label for lot-specific concentration
      Preservative≤0.1% sodium azide
      CommentsTwo p73 isoforms are expressed in most cells. p73α migrates as an ~80 kDa protein and p73β migrates as a ~68 kDa protein. This antibody detects both p73α and p73β in immunoblots and will immunoprecipitate p73α, p73β, and p73α/p73β complexes. This antibody will also supershift p73α/DNA and p73β/DNA complexes. For gel shift, use Cat. No. OP109L and resuspend in 100 µl buffer. For blotting, use 150 to 300 µg total cell lysate per lane. Antibody should be titrated for optimal results in individual systems.

      1. Using fresh lysates, display 150-300 µg of total cellular protein on 10% PAGE/SDS. Transfer to nitrocellulose.
      2. Block nitrocellulose for 30 min at RT in TBS/T (10 mM Tris-HCl, pH 8.0, 150 mM NaCl, 0.2% Tween®-20 detergent) buffer containing 5% non-fat dry milk and 1% normal goat serum.
      3. Incubate blot with primary antibody in TBS/T (10 mM Tris-HCl, pH 8.0, 150 mM NaCl, 0.2% Tween®-20 detergent) for 2 h at RT or overnight at 4°C.
      4. Wash blot 3 times for 10 min. each with TBS/T.
      5. Incubate blot with goat anti-mouse IgG conjugated to HRP at 50 ng/mL in TBS/T for 30 min at RT.
      6. Wash blot 3 times for 10 min. each with TBS/T.
      7. Detect bound antibodies using chemiluminescence.
      8. Multiple bands may appear in all lanes due to non-specific cross-reacting material. However, bands corresponding to p73α and p73β should be seen in 293 but not in SK-N-AS lysates.
      Storage +2°C to +8°C
      Do Not Freeze Yes
      Toxicity Standard Handling
      ReferencesDi Como, C.J., et al. 1999. Mol. Cell Biol. 19, 1438.
      De Laurenzi, V., et al. 1998. J. Exp. Med. 188, 1763.
      Dobbelstein, M. and J. Roth. 1998. J. Gen. Virol. 79, 3079.
      Higashino, F., et al. 1998. Proc. Natl. Acad. Sci. USA 95, 15683.
      Marin, M.C., et al. 1998. Mol. Cell Biol. 18, 6316.
      Prabhu, N.S., et al. 1998. Int. J. Oncol. 13, 5.
      Roth, J., et al. 1998. J. Virol. 72, 8510.
      Zhu, J., et al. 1998. Cancer Res 58, 5061.
      Jost, C.A., et al. 1997. Nature 389, 191.
      Kaghad, M., et al. 1997. Cell 90, 809.
      Application referencesOriginal Clone Marin, M.C., et al. 1998. Mol. Cell Biol. 18, 6316.