CD137 antibody (AA 101-200)

Catalog No. ABIN740445

Product Details anti-CD137 Antibody

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

Binding Specificity: AA 101-200

Reactivity: Human, Mouse, Rat

Host: Rabbit

Clonality: Polyclonal

Conjugate: This CD137 antibody is un-conjugated

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

Cross-Reactivity: Human, Mouse, Rat

Purification: Purified by Protein A.

Immunogen: KLH conjugated synthetic peptide derived from human CD137

Isotype: IgG

Application Details

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

Restrictions: For Research Use only

Independent Validation (FACS)

Validation #101793 (Flow Cytometry)

by: Pharmacology Department, Faculty of Medicine, Granada University

No.: #101793

Date: 09/13/2017

Antigen: CD137

Lot Number: AG05152084

Method validated: Flow Cytometry

Positive Control: rat adipose-derived mesenchymal stem cells

Negative Control: cells stained with secondary antibody only

Notes:

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

Validation Images

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.

Full Methods

Primary Antibody: ABIN740445

Secondary Antibody: 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).

Experimental Notes:

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.

Handling

Format: Liquid

Concentration: 1 μg/μL

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

Preservative: ProClin

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

Storage: 4 °C,-20 °C

Storage Comment: Shipped at 4°C. Store at -20°C for one year. Avoid repeated freeze/thaw cycles.

Expiry Date: 12 months

References for anti-CD137 antibody (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).

Target Details for CD137

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

Alternative Name: CD137 (TNFRSF9 Products)

Background:

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.

Gene ID: 3604

UniProt: Q07011

Pathways: Cancer Immune Checkpoints