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Anti-Human 4-1BB (4-1BB)
Purified No Carrier Protein
(TNFRSF9, CD137 Antigen, T-Cell Antigen ILA )
Monoclonal Antibody
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Prod. No.:B591
Clone:145501
Isotype:Mouse IgG2b
Pkg. Size:500 µg, 1 mg
Storage:-20°C to -70°C Detailed storage instructions below.
Description
Specificity:
Mouse Anti-Human 4-1BB (Clone 145501) recognizes an epitope on Human 4-1BB. This monoclonal antibody was purified using multi-step affinity chromatography methods such as Protein A or G depending on the species and isotype.

Background:
4-1BB (CD137) is a costimulatory molecule expressed on the surface of activated T cells1 and plays a role in T-cell-mediated proliferative response.2 4-1BB signal regulates CD28 co-stimulation in the targeted subset cells to favor Th1 development and maintain long-term cell growth.3 4-1BB is also expressed on dandritic cells (DCs) and controls the duration, DC-T interaction, and, therefore, immunogenicity.4

Host Species
Mouse
Immunogen
NS0-Derived Recombinant Human 4-1BB Extracellular Domain
Formulation
This monoclonal antibody has been 0.2 µm filtered and lyophilized from modified Dulbecco's phosphate buffered saline (1X PBS) pH 7.2 - 7.3 containing 5.0% w/v trehalose with no calcium, magnesium or preservatives present.
Reconstitution
This lyophilized antibody may be aseptically reconstituted with sterile PBS to a final volume of 1.0 ml. This will result in a final antibody concentration of 500 µg/ml.
Storage and Stability
The lyophilized antibody can be stored desiccated at -20°C to -70°C for up to twelve months. The reconstituted antibody can be stored for at least four weeks at 2-8°C. For long-term storage of the reconstituted antibody, aseptically aliquot into working volumes and store at -20°C to -70°C in a manual defrost freezer. Avoid repeated freeze thaw cycles. No detectable loss of activity was observed after six months.
Reported Applications
Indirect ELISA, FC, WB
Indirect ELISA: This antibody can be used in an indirect detection ELISA at 0.5 - 1.0 µg/ml with a suitable second step reagent such as Goat Anti-Mouse IgG (H&L)-Horseradish Peroxidase (Leinco Prod. No.: M114). The detection sensitivity of this indirect ELISA for Human 4-1BB is approximately 10 ng/well.
Flow Cytometry: It is recommended to use the indirect method for signal enhancement when enumerating cells expressing 4-1BB. A suggested method would be to stain cells expressing 4-1BB with 0.25 µg per 1 - 2.5 x 105 cells in a total staining volume of ≤200 µl followed by Goat Anti-Mouse IgG (H&L)-R-phycoerythrin (Leinco Prod. No.: M1189).
Western Blotting: To detect Human 4-1BB this monoclonal antibody can be used at a concentration of 1-2 µg/ml. This monoclonal antibody should be used in conjunction with compatible second-step reagents such as Goat Anti-Mouse IgG (H&L)-Horseradish Peroxidase (Leinco Prod. No.: M114) and a chromogenic substrate such as TMB HRPO Membrane/IHC Substrate UltraSensitive™ One Component "Ready Use" (Leinco Prod. No.: T343). The detection limit for Human 4-1BB is 25 ng/lane under either reducing or non-reducing conditions. The sensitivity of detection may increase up to 50 fold when a chemiluminescent substrate is used.
Each investigator should determine their own optimal working dilution for specific applications.


Country of Origin
USA
References
1. Shan B et al. (2008) Cell Mol Immunol. 5: 379
2. Kwan BS et al. (1996) Cell Immunol. 169: 91
3. Kwan BS et al. (1998) Eur J Immunol. 28: 881
4. Kwan BS et al. (2009) J Immunol. 182: 4107
Related Products
Goat Anti-Mouse IgG (H&L)-Horseradish Peroxidase (Leinco Prod. No.: M114)
Goat Anti-Mouse IgG (H&L)-R-phycoerythrin (Leinco Prod. No.: M1189)
TMB HRPO Membrane/IHC Substrate UltraSensitive™ One Component "Ready Use" (Leinco Prod. No.: T343)
Anti-Mouse 4-1BB (Antigen Aff. Pur.) (Leinco Prod. No.: B579)
Anti-Mouse 4-1BB-Biotin (Antigen Aff. Pur.) (Leinco Prod. No.: B561)
Products are for research use only. Not for use in diagnostic or therapeutic procedures.
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