CD137 Antikörper (AA 101-200)

Produktnummer ABIN740445

Produktdetails anti-CD137 Antikörper

Target: CD137 (TNFRSF9) (Tumor Necrosis Factor Receptor Superfamily, Member 9 (TNFRSF9))

Bindungsspezifität: AA 101-200

Reaktivität: Human, Maus, Ratte

Wirt: Kaninchen

Klonalität: Polyklonal

Konjugat: Dieser CD137 Antikörper ist unkonjugiert

Applikation: ELISA, Flow Cytometry (FACS), Western Blotting (WB), Immunohistochemistry (Paraffin-embedded Sections) (IHC (p))

Kreuzreaktivität: Human, Maus, Ratte

Aufreinigung: Purified by Protein A.

Immunogen: KLH conjugated synthetic peptide derived from human CD137

Isotyp: IgG

Anwendungsinformationen

Applikationshinweise: WB 1:300-5000
ELISA 1:500-1000
FCM 1:20-100
IHC-P 1:200-400

Beschränkungen: Nur für Forschungszwecke einsetzbar

Independent Validation (FACS)

Validierung #101793 (Flow Cytometry)

by: Pharmacology Department, Faculty of Medicine, Granada University

No.: #101793

Datum: 13.09.2017

Antigen: CD137

Chargennummer: AG05152084

Validierte Anwendung: Flow Cytometry

Positivkontrolle: rat adipose-derived mesenchymal stem cells

Negativkontrolle: cells stained with secondary antibody only

Bewertung:

Passed. ABIN740445 specifically recognizes Cd137 in rat adipose-derived mesenchymal stem cells.

Validierungsbilder

Flow cytometric analysis of cells stained with ABIN740445 and an AF680-conjugated secondary antibody (A) or the secondary antibody only (B). The top row illustrates the gating for life cells. The second and third row show dots blots and histograms respectively of the positively stained cell populations.

Protokoll

Primärantikörper: ABIN740445

Sekundärantikörper: donkey anti-rabbit Alexa Fluor 680 conjugated antibody (ThermoFisher Scientific, A10043, lot 1812322)

Full Protocol:

• Grow rat adipose-derived mesenchymal stem cells in DMEM high glucose, GLUTAMAX, pyruvate supplemented (Gibco, 31966021, lot 1774921) supplemented with 10% fetal bovine serum (Gibco, 16000044, lot 1774921) 1% penicillin/streptomycin (Gibco, 15140122, lot 1759446), at 37°C and 5% CO₂ in ml on a flask to 95% confluency.
• Wash cells with PBS buffer.
• Detach cells from the flask with trypsin-versene solution (Lonza, H3BE17-161E, lot RB35937) for 3min at 37°C and 5% CO₂.
• Centrifuge cells for at 400xg for 5min at RT, aspirate the supernatant.
• Resuspend cells in 1mL of 1% BSA in PBS per 2x10⁶ cells and incubate 20min at RT.
• Incubate cells with primary rabbit anti-CD137 antibody (antibodies-online, ABIN740445, lot AG05152084) diluted 1:35 in 1% BSA in PBS buffer for 20min on ice. Include a negative control without primary antibody.
• Wash cells 1x with PBS buffer and centrifuge them for 400xg for 5min at RT.
• Resuspend cells in 175μl 1% BSA in PBS buffer.
• Incubate cells with secondary donkey anti-rabbit Alexa Fluor 680 conjugated antibody (ThermoFisher Scientific, A10043, lot 1812322) diluted 1:200 in buffer for 15min in ice.
• Wash cells 2xwith PBS buffer and centrifuge them for 400xg for 5min at RT.
• Resuspend the pellet in 500μl PBS buffer.
• Data acquisition in a BD FACSCanto II flow cytometer, equipped with two laser lines of 488nm and 633nm and two scattered light detectors (FSC and SSC). Detect fluorescence in PMTs with filters suitable for the fluorescence of the fluorophore AF680 (735LP and 780/60BP). Threshold is established in FSC. The data is saved in files in FCS format and analyzed offline with the FACSDiva software (BD).

Anmerkungen:

ABIN740445 reveals a specific fluorescence signal in rat adipose-derived mesenchymal stem cells. This is the first time that flow cytometry is performed in this type of cells using this antibody. No signal was registered in the negative control cells.

Handhabung

Format: Liquid

Konzentration: 1 μg/μL

Buffer: 0.01M TBS( pH 7.4) with 1 % BSA, 0.02 % Proclin300 and 50 % Glycerol.

Konservierungsmittel: ProClin

Vorsichtsmaßnahmen: This product contains ProClin: a POISONOUS AND HAZARDOUS SUBSTANCE, which should be handled by trained staff only.

Lagerung: 4 °C,-20 °C

Informationen zur Lagerung: Shipped at 4°C. Store at -20°C for one year. Avoid repeated freeze/thaw cycles.

Haltbarkeit: 12 months

Referenzen für anti-CD137 Antikörper (ABIN740445)

Claycombe, Vomhof-DeKrey, Garcia, Johnson, Uthus, Roemmich et al.: "Decreased beige adipocyte number and mitochondrial respiration coincide with increased histone methyl transferase (G9a) and reduced FGF21 gene expression in Sprague-Dawley rats fed prenatal low ..." in: The Journal of nutritional biochemistry, Vol. 31, pp. 113-21, (2016) (PubMed).

Daquinag, Tseng, Salameh, Zhang, Amaya-Manzanares, Dadbin, Florez, Xu, Tong, Kolonin: "Depletion of white adipocyte progenitors induces beige adipocyte differentiation and suppresses obesity development." in: Cell death and differentiation, (2014) (PubMed).

Details zu CD137

Target: CD137 (TNFRSF9) (Tumor Necrosis Factor Receptor Superfamily, Member 9 (TNFRSF9))

Andere Bezeichnung: CD137 (TNFRSF9 Produkte)

Synonyme: TNFRSF9 antikoerper, zgc:136557 antikoerper, 4-1BB antikoerper, CD137 antikoerper, CDw137 antikoerper, ILA antikoerper, A930040I11Rik antikoerper, AA408498 antikoerper, AI325004 antikoerper, Cd137 antikoerper, Ly63 antikoerper, tumor necrosis factor receptor superfamily member 9 antikoerper, TNF receptor superfamily member 9 antikoerper, tumor necrosis factor receptor superfamily, member 9a antikoerper, tumor necrosis factor receptor superfamily, member 9 antikoerper, TNFRSF9 antikoerper, tnfrsf9a antikoerper, Tnfrsf9 antikoerper

Hintergrund:

Synonyms: ILA, 4-1BB, CD137, CDw137, Tumor necrosis factor receptor superfamily member 9, 4-1BB ligand receptor, T-cell antigen 4-1BB homolog, T-cell antigen ILA, TNFRSF9

Background: Receptor for TNFSF9/4-1BBL. Possibly active during T cell activation.

Gen-ID: 3604

UniProt: Q07011

Pathways: Cancer Immune Checkpoints