TNFRSF8 antibody

Catalog No. ABIN492585

Product Details anti-TNFRSF8 Antibody

Target: TNFRSF8 (Tumor Necrosis Factor Receptor Superfamily, Member 8 (TNFRSF8))

Reactivity: Human

Host: Mouse

Clonality: Monoclonal

Conjugate: This TNFRSF8 antibody is un-conjugated

Application: Flow Cytometry (FACS), Western Blotting (WB), Immunohistochemistry (Paraffin-embedded Sections) (IHC (p)), Immunoprecipitation (IP)

Specificity: This antibody reacts with CD30 antigen (120 kDa) on Western blotting, Immunoprecipitation, and Flow Cytometry.

Cross-Reactivity (Details): Species reactivity (tested):Human.

Characteristics: Synonyms: TNFRSF8, D1S166E, CD30L receptor, KI-1 antigen, Tumor necrosis factor receptorsuperfamily member 8, Lymphocyte activation antigen CD30

Purification: Protein-A Agarose Chromatography of hybridoma supernatant.

Immunogen: Co cell line cellsRemarks: Hybridoma was established by fusion of mouse myeloma cell with Balb/c mousesplenocyte

Clone: Ber-H2

Isotype: IgG1

Application Details

Application Notes: Western blotting: 1 μg/mL for chemiluminescence detection system. Positive Control: CCRF-CEM. Immunoprecipitation: 2 μg/200 μL of cell extract from 5x10^6 cells. Flow Cytometry: 10 μg/mL (final concentration). Immunohistochemistry on Paraffin Embedded Section: 1-5 μg/mL (Heat treatment isnecessary). Microwave oven, 2 times for 10 minutes each in 10 mM Citrate buffer ( pH 6.5). Detailed procedure is provided in Protocols.
Other applications not tested.
Optimal dilutions are dependent on conditions and should be determined by the user.

Protocol: SDS-PAGE & Western Blotting1) Wash the cells 3 times with PBS and suspend with 10 volume of cold Lysis buffer (50 mMTris-HCl, pH 7. 2, 250 mM NaCl, 0. 1% NP-40, 2 mM EDTA, 10%glycerol) containingappropriate protease inhibitors. Incubate it at 4°C with rotating for 30 minutes, thensonicate briefly (up to 10 seconds). 2) Centrifuge the tube at 12,000 x g for 10 minutes at 4°C and transfer the supernatant toanother tube. Measure the protein concentration of the supernatant and add the cold Lysisbuffer to make 8 mg/mL solution. 3) Mix the sample with equal volume of Laemmli's sample buffer. 4) Boil the samples for 3 minutes and centrifuge. Load 10 μL of the sample per lane in a 1mm thick SDS-polyacrylamide gel for electrophoresis. 5) Blot the protein to a polyvinylidene difluoride (PVDF) membrane at 1 mA/cm2 for 1 hourin a semi-dry transfer system (Transfer Buffer: 25 mM Tris, 190 mM glycine, 20% MeOH). See the manufacture's manual for precise transfer procedure. 6) To reduce nonspecific binding, soak the membrane in 10% skimmed milk (in PBS, pH7. 2) for 1 hour at room temperature, or overnight at 4°C. 7) Incubate the membrane with primary antibody diluted with PBS, pH 7. 2 containing 1%skimmed milk as suggest in the APPLICATIONS for 1 hour at room temperature. (Theconcentration of antibody will depend on condition. )8) Wash the membrane with PBS-T [0. 05% Tween-20 in PBS] (5 minutes x 3 times). 9) Incubate the membrane with the 1: 10,000 HRP-conjugated anti-mouse IgG diluted with1% skimmed milk (in PBS, pH 7. 2) for 1 hour at RT. 10) Wash the membrane with PBS-T (10 minutes x 3 times). 11) Wipe excess buffer on the membrane, then incubate it with appropriatechemiluminescence reagent for 1 minute. 12) Remove extra reagent from the membrane by dabbing with paper towel, and seal it inplastic wrap. 13) Expose to an X-ray film in a dark room for 3 minutes. 14) Develop the film as usual. The condition for exposure and development may vary. Positive Control: U251, CCRF-CEM. Immunoprecipitation1) Wash the cells 3 x with PBS and suspend with 10 volume of cold Lysis buffer (50 mMTris-HCl pH 7. 2, 250 mM NaCl, 0. 1% NP-40, 2 mM EDTA, 10% glycerol) containingappropriate protease inhibitors. Incubate it at 4°C with rotating for 30 minutes, thensonicate briefly (up to 10 seconds). 2) Centrifuge the tube at 12,000 x g for 10 minutes at 4°C and transfer the supernatant toanother tube.

Restrictions: For Research Use only

Handling

Concentration: 1.0 mg/mL

Buffer: PBS, pH 7.2 containing 50 % Glycerol without preservatives.

Preservative: Without preservative

Storage: -20 °C

Storage Comment: Store the antibody (in aliquots) at -20 °C. Avoid repeated freezing and thawing.
Shelf life: one year from despatch.

Expiry Date: 12 months

Target Details for TNFRSF8

Target: TNFRSF8 (Tumor Necrosis Factor Receptor Superfamily, Member 8 (TNFRSF8))

Alternative Name: CD30 (TNFRSF8 Products)

Background: CD30, also known as Ki-1, TNFRSF8, or Be-H2, is a 120 kDa glycoprotein expressed on the surface of mitogen-activated B-cells and T-cells but not on resting lymphocytes or monocytes. CD30 is also a marker for Hodgkin and Sternberg-Reed cells of Hodgkin's lymphomas and related hematologic malignancies. Soluble forms of CD30 have been found in the serum of patients with adult T-cell leukemia or other CD30+ lymphomas. The CD30 ligand, CD153, is a type II transmembrane glycoprotein that enhances proliferation of activated T-cells and induces apoptosis in CD30+ lymphoma-derived cell lines.Synonyms: CD30L receptor, D1S166E, KI-1 antigen, Lymphocyte activation antigen CD30, TNFRSF8, Tumor necrosis factor receptor superfamily member 8

Gene ID: 943

UniProt: P28908