Flow Cytometry. Other applications not tested. Optimal dilutions are dependent on conditions and should be determined by the user.
Protocol
FLOW CYTOMETRY ANALYSIS: Method: 1. Prepare a cell suspension in media A. For cell preparations, deplete the red blood cellpopulation with Lympholyte®-M cell separation medium. 2. Wash 2 times. 3. Resuspend the cells to a concentration of 2x10e7 cells/ml in media A. Add 50 µl of thissuspension to each tube (each tube will then contain 1 x 10e6 cells, representing 1 test). 4. To each tube, add ~1. 0 µg* of this Ab per 10e6 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 areprotected 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 propidiumiodide at 0. 5 mg/ml in PBS. This stains dead cells by intercalating in DNA.
Restrictions
For Research Use only
Concentration
0.1 mg/mL
Buffer
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.
Preservative
Sodium azide
Precaution of Use
This product contains sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
Storage
4 °C
Storage Comment
Store the antibody undiluted at 2-8 °C. DO NOT FREEZE!
Target
TNFRSF1A
(Tumor Necrosis Factor Receptor Superfamily, Member 1A (TNFRSF1A))
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: TNF-R1, TNF-RI, TNFR-I, Tnfrsf1a, Tumor necrosis factor receptor 1, Tumor necrosis factor receptor superfamily member 1A, Tumor necrosis factor receptor type I, p55, p60