Tnfrsf1a Rat Monoclonal Antibody [Clone ID: HM104]

CAT#: CL044R

Tnfrsf1a rat monoclonal antibody, clone HM104, PE

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Specifications

Product Data

• Clone Name: HM104
• Applications: FC
• Recommended Dilution: Flow Cytometry.
• Reactivities: Mouse
• Host: Rat
• Isotype: IgG2a
• Clonality: Monoclonal
• Specificity: This Antibody reacts with the extracellular part of the Mouse TNF-Receptor (p55) and with the soluble receptor.
• Formulation: PBS containing 0.09% Sodium Azide as preservative and EIA grade BSA as a stabilizing protein to bring total protein concentration to 4-5 mg/ml.
  Label: PE
  State: Liquid purified IgG fraction
  Label: R-Phycoerythrin
• Concentration: 0.1 mg/ml
• Conjugation: PE
• Gene Name: tumor necrosis factor receptor superfamily, member 1a
• Database Link:
  Entrez Gene 21937 Mouse
• Background: Tumor Necrosis Factor (TNF) is a cytokine whose function is mediated through two distinct cell surface receptors (TNF Receptor I and TNF Receptor II) that are included in the TNF Receptor superfamily along with FAS antigen and CD40. TNF Receptors I and II are 55 and 75 kDa members, respectively, of a family of cell surface molecules including nerve growth factor receptor, Fas/Apo1, CD30, OX40, and 41BB, which are characterized by cysteine rich motifs in the extracellular domain. While TNF Receptor I and TNF Receptor II share 28% sequence homology in the extracellular domains, their intracellular domains lack sequence homology, suggesting that they differ in their internal signal transduction pathways. TNF Receptor I contains an approximately 80 amino acid death domain near its carboxy terminus capable of transmitting an apoptotic signal through its interaction with TRADD (TNF Receptor I associated death domain protein), and subsequent interactions with FADD. TNF Receptor I can also activate the transcription factor NFkB via TRAF2 (TNF Receptor associated factor 2). The cytoplasmic domain of TNF Receptor I can directly interact with Jak kinase, thereby activating the JAK/STAT signal transduction cascade.
  TNF Receptor I is expressed by virtually all nucleated mammalian cells, including hepatocytes, monocytes and neutrophils, cardiac muscle cells, endothelial cells, and CD34 + hematopoietic progenitors. Both TNF alpha and TNF beta bind to TNF Receptor I.
• Synonyms: Tumor necrosis factor receptor 1, TNF-R1, TNF-RI, TNFR-I, p55, p60, Tnfrsf1a
• Note: Protocol: FLOW CYTOMETRY ANALYSIS:
  Method:
  1. Prepare a cell suspension in media A. For cell preparations, deplete the red blood cell population with Lympholyte®-M cell separation medium.
  2. Wash 2 times.
  3. Resuspend the cells to a concentration of 2x10⁷ cells/ml in media A. Add 50 µl of this suspension to each tube (each tube will then contain 1 x 10⁶ cells, representing 1 test).
  4. To each tube, add ~1.0 µg* of this Ab per 10⁶ cells.
  5. Vortex the tubes to ensure thorough mixing of antibody and cells.
  6. Incubate the tubes for 30 minutes at 4°C. (It is recommended that the tubes are protected from light, since most fluorochromes are light sensitive.)
  7. Wash 2 times at 4°C.
  8. Resuspend the cell pellet in 50 µl ice cold media B.
  9. Transfer to suitable tubes for flow cytometric analysis containing 15 µl of propidium iodide at 0.5 mg/ml in PBS. This stains dead cells by intercalating in DNA.
  
  Media:
  A. Phosphate buffered saline (pH 7.2) + 5% normal serum of host species + Sodium Azide (100 µl of 2M Sodium Azide in 100 mls).
  B. Phosphate buffered saline (pH 7.2) + 0.5% Bovine Serum Albumin + Sodium Azide (100 µl of 2M Sodium Azide in 100 mls).
  
  Tissue Distribution by Flow Cytometry Analysis:
  Mouse Strain: CBA
  Cell Concentration: 1x10⁶ cells per test
  Antibody Concentration Used: 1.0 µg/10⁶ cells
  Isotypic Control: PE Rat IgG2a
  Cell Source: Spleen
  Percentage of Cells Stained Above Control: 7.10%