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Purified Mouse Anti-Human p300
Purified Mouse Anti-Human p300

Immunoprecipitation of p300 and coprecipitation of E1A with NM11 (Cat. No. 554215). NM11 immunoprecipitated p300 from Cos-7 cells (lane 1), and both p300 and E1A from Cos-7 cells transfected with plasmids encoding either adenovirus (Ad)12 13S E1A (lane 2) or Ad12 12S E1A (lane 3) Cos-7 cells [35]S labeled.

Immunoprecipitation of p300 and coprecipitation of E1A with NM11 (Cat. No. 554215). NM11 immunoprecipitated p300 from Cos-7 cells (lane 1), and both p300 and E1A from Cos-7 cells transfected with plasmids encoding either adenovirus (Ad)12 13S E1A (lane 2) or Ad12 12S E1A (lane 3) Cos-7 cells [35]S labeled.

Product Details
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BD Pharmingen™
Human, Mouse, Rat, Monkey (Tested in Development)
Mouse IgG2b
Human p300
Western blot (Routinely Tested), Dot Blot, Immunoprecipitation (Reported)
300 kDa
0.5 mg/ml
AB_395310
Aqueous buffered solution containing ≤0.09% sodium azide.
RUO


Preparation And Storage

The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. Store undiluted at 4°C.

Recommended Assay Procedures

Applications include Western blot and immunoprecipitation. Clone NMII is routinely tested by western blot analysis using the transfer protocol for high molecular weight proteins, please refer to http://www.bdbiosciences.com/pharmingen/protocols/Western_Blotting.shtml.  Ad5 transformed cell lines including 293 cells (ATCC CRL 1573) are suggested as positive controls.

Product Notices

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  2. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  3. Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
554215 Rev. 9
Antibody Details
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NM11

Many serotypes of human adenoviruses (Ad) are able to transform rodent cells in culture because of functions encoded by early region adenovirus 1A (E1A) and 1B (E1B) genes. These virus early gene products bind to specific cellular proteins that normally act to restrict cell growth. The E1A gene, essential for viral replication, encodes protein products with three amino acid regions (1, 2 and 3) conserved among all known adenovirus serotypes. Regions 1 and 2 and the relatively unconserved amino terminus (N) are E1A sequences required for cell growth-regulating functions. The region 2 site binds the retinoblastoma (Rb) and Rb related proteins, p107 and p130. The amino terminal site binds p300, a large cellular DNA binding protein. p300 is a relatively stable, ubiquitously expressed, nuclear phosphoprotein which is conserved among a variety of mammalian species. It is actively synthesized and phosphorylated in both quiescent and proliferating cells. A phosphatase sensitive form with decreased mobility has been identified in M-phase enriched cell populations. Evidence suggests that p300 plays a role in cellular transcription mechanisms. It has sequence-specific DNA binding activity with a consensus DNA binding sequence similar to enhancer elements targeted by the E1A repressor function. The presence of p300 and p300-associated proteins in complexes with the TATA binding protein suggests that p300 has specific interactions with basal transcription mechanisms. Through its association with the TATA regions, p300 may also play a role in the ability of E1A to activate the hsp70 promoter dependent on specific TATA box sequences. p300 migrates at a reduced molecular weight at ~300 kD.

Clone NM11 recognizes human p300 a cellular E1A binding protein p300. Specifically, it has been shown to recognize p300 from human, monkey, mouse and rat cell lysates. NM11 will also coprecipitate E1A in association with p300 when E1A is present. Affinity purified p300 was used as immunogen. p300 was affinity purified from 293 human kidney cells (adenovirus type 5 [Ad5] transformed cell line) by co-immunoprecipitation using an E1A-specific monoclonal antibody (M73), which was covalently linked to protein A-Sepharose beads.

554215 Rev. 9
Format Details
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Purified
Tissue culture supernatant is purified by either protein A/G or affinity purification methods. Both methods yield antibody in solution that is free of most other soluble proteins, lipids, etc. This format provides pure antibody that is suitable for a number of downstream applications including: secondary labeling for flow cytometry or microscopy, ELISA, Western blot, etc.
Purified
554215 Rev.9
Citations & References
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Development References (3)

  1. Rikitake Y, Moran E. DNA-binding properties of the E1A-associated 300-kilodalton protein. Mol Cell Biol. 1992; 12(6):2826-2836. (Biology). View Reference
  2. Wang HG, Yaciuk P, Ricciardi RP, Green M, Yokoyama K, Moran E. The E1A products of oncogenic adenovirus serotype 12 include amino-terminally modified forms able to bind the retinoblastoma protein but not p300. J Virol. 1993; 67(8):4804-4813. (Clone-specific: Immunoprecipitation). View Reference
  3. Yaciuk P, Moran E. Analysis with specific polyclonal antiserum indicates that the E1A-associated 300-kDa product is a stable nuclear phosphoprotein that undergoes cell cycle phase-specific modification. Mol Cell Biol. 1991; 11(11):5389-5397. (Immunogen). View Reference
554215 Rev. 9

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Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described


Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims.  Comparisons are not made against non-BD technologies, unless otherwise noted.

For Research Use Only. Not for use in diagnostic or therapeutic procedures.

Refer to manufacturer's instructions for use and related User Manuals and Technical Data Sheets before using this product as described.

Comparisons, where applicable, are made against older BD technology, manual methods or are general performance claims. Comparisons are not made against non-BD technologies, unless otherwise noted.