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      Purified Mouse Anti-Nur77
      Purified Mouse Anti-Nur77
      Left Figure: Western blot analysis of Nur77 expression on mouse thymocytes. Thymocytes were stimulated with PMA (20 ng/ml) and ionomycin (500 ng/ml at 37°C for 2 hr). Lysates were prepared and separated by SDS/PAGE. Blots were probed with Purified Mouse Anti-Nur77 (Cat. No. 554088) at concentrations of 2.0 (lane 1), 1.0 (lane 2), and 0.5 µg/ml (lane 3). Visualization was carried out with HRP Goat Anti-Mouse Ig (Cat. No. 554002). Nur77 is detected as a protein of ~77 kDa. Right Figure: Flow cytometric analysis of Nur77 expression on stimulated mouse thymocytes. Thymocytes from C57/BL/6 mice were left unstimulated (gray line histogram) or stimulated (black line histograms) with PMA (20ng/ml) and  Ionomycin (500ng/ml) for 2 hours and then stained intracellularly with Purified Mouse IgG1, k Isotype Control (Cat.No. 554121; black dashed line) or Purified Mouse Anti-Nur77 antibody (black solid line) at 0.125ug/test using BD Pharmingen™ Transcription Factor Buffer Set (Cat. No. 562574). Purified antibody was followed by PE conjugated Goat Anti-Mouse Ig (Multiple Absorption Secondary antibody (Cat.No. 550589). Histogram plots were derived from gated events with the forward and side light-scatter characteristics of viable cells. Flow cytometric analysis was performed using a BD LSRFortessa™ X-20 Flow Cytometer System. Data shown on this Technical Data Sheet are not lot specific.
      Purified Mouse Anti-Nur77
      Left Figure: Western blot analysis of Nur77 expression on mouse thymocytes. Thymocytes were stimulated with PMA (20 ng/ml) and ionomycin (500 ng/ml at 37°C for 2 hr). Lysates were prepared and separated by SDS/PAGE. Blots were probed with Purified Mouse Anti-Nur77 (Cat. No. 554088) at concentrations of 2.0 (lane 1), 1.0 (lane 2), and 0.5 µg/ml (lane 3). Visualization was carried out with HRP Goat Anti-Mouse Ig (Cat. No. 554002). Nur77 is detected as a protein of ~77 kDa. Right Figure: Flow cytometric analysis of Nur77 expression on stimulated mouse thymocytes. Thymocytes from C57/BL/6 mice were left unstimulated (gray line histogram) or stimulated (black line histograms) with PMA (20ng/ml) and  Ionomycin (500ng/ml) for 2 hours and then stained intracellularly with Purified Mouse IgG1, k Isotype Control (Cat.No. 554121; black dashed line) or Purified Mouse Anti-Nur77 antibody (black solid line) at 0.125ug/test using BD Pharmingen™ Transcription Factor Buffer Set (Cat. No. 562574). Purified antibody was followed by PE conjugated Goat Anti-Mouse Ig (Multiple Absorption Secondary antibody (Cat.No. 550589). Histogram plots were derived from gated events with the forward and side light-scatter characteristics of viable cells. Flow cytometric analysis was performed using a BD LSRFortessa™ X-20 Flow Cytometer System. Data shown on this Technical Data Sheet are not lot specific.
      Product Details
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      BD Pharmingen™
      Nr4a1; Nur77; NGFI-B; GFRP1; NGFIB; N10; TR3; Hbr-1; TIS1
      Mouse (QC Testing), Human (Tested in Development)
      Mouse IgG1
      Full-length Mouse Nur77 fusion protein
      Western blot (Routinely Tested), Flow cytometry (Tested During Development)
      67-88 kDa
      0.5 mg/ml
      AB_395232
      Aqueous buffered solution containing ≤0.09% sodium azide.
      RUO


      Preparation And Storage

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

      Mouse thymocytes treated with PMA and ionomycin are suggested as a positive control.

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      Product Notices

      1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
      2. An isotype control should be used at the same concentration as the antibody of interest.
      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.
      4. Sodium azide is a reversible inhibitor of oxidative metabolism; therefore, antibody preparations containing this preservative agent must not be used in cell cultures nor injected into animals. Sodium azide may be removed by washing stained cells or plate-bound antibody or dialyzing soluble antibody in sodium azide-free buffer. Since endotoxin may also affect the results of functional studies, we recommend the NA/LE (No Azide/Low Endotoxin) antibody format, if available, for in vitro and in vivo use.
      5. Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
      6. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
      7. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
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      554088 Rev. 11
      Antibody Details
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      12.14

      The 12.14 monoclonal antibody specifically recognizes Nur77, a zinc-finger transcription factor also known as Nerve growth factor IB (NGFI-B), Testicular receptor 3 (TR3), or Nuclear protein N10. Nur77 is encoded by Nr4a1 (Nuclear receptor subfamily 4 group A member 1), an immediate-early response gene that belongs to the nuclear receptor superfamily. This inducible orphan nuclear receptor is comprised of an N-terminal transactivation domain, followed by a central DNA-binding domain and a putative C-terminal ligand-binding domain for which no ligand has been identified. In electrophoretic analyses, Nur77 migrates as diffuse protein bands between 67 and 88 kDa depending on its level of phosphorylation or other post-translational modifications. Nr4a1 expression can be rapidly induced by diverse stimuli in cells from primary and secondary lymphoid tissues and other tissues including the brain, muscle, ovary, and testis. Nur77 expression is rapidly upregulated by antigen-stimulated mouse thymocytes and may promote activation- induced apoptosis during negative selection. Ag receptor-mediated signaling by B cells, T cells, or T cell hybridomas can also lead to rapid upregulated Nur77 expression which may regulate cellular proliferation and survival. In response to certain growth factors, cytokines, inflammatory mediators, or stress-inducing stimuli, other cell types, including myeloid or stromal cell types, can upregulate Nur77 expression which affects their growth, differentiation, proliferation or survival. Clone 12.14 recognizes mouse Nur77 and reportedly crossreacts with human Nur77.

      554088 Rev. 11
      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
      554088 Rev.11
      Citations & References
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      Development References (5)

      1. Davis IJ, Hazel TG, Chen RH, Blenis J, Lau LF. Functional domains and phosphorylation of the orphan receptor Nur77. Mol Endocrinol. 1993; 7(8):953-964. (Biology). View Reference
      2. Davis IJ, Lau LF. Endocrine and neurogenic regulation of the orphan nuclear receptors Nur77 and Nurr-1 in the adrenal glands. Mol Cell Biol. 1994; 14(5):3469-3483. (Biology). View Reference
      3. Hazel TG, Misra R, Davis IJ, Greenberg ME, Lau LF. Nur77 is differentially modified in PC12 cells upon membrane depolarization and growth factor treatment. Mol Cell Biol. 1991; 11(6):3239-3246. (Biology). View Reference
      4. Liu ZG, Smith SW, McLaughlin KA, Schwartz LM, Osborne BA. Apoptotic signals delivered through the T-cell receptor of a T-cell hybrid require the immediate-early gene nur77. Nature. 1994; 367(6460):281-284. (Biology). View Reference
      5. Woronicz JD, Calnan B, Ngo V, Winoto A. Requirement for the orphan steroid receptor Nur77 in apoptosis of T-cell hybridomas. Nature. 1994; 367(6460):277-281. (Biology). View Reference
      View All (5) View Less
      554088 Rev. 11

      Please refer to Support Documents for Quality Certificates


      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.

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