Goat Anti-Mouse 4-1BB
Antigen Affinity Purified
(TNFRSF9, CD137 Antigen, T-Cell Antigen ILA )
Polyclonal Antibody
| Prod. No.: | B579 |
| Host: | Goat |
| Pkg. Size: | 1 mg, 100 µg |
| Storage: | -20°C to -70°C Detailed storage instructions below. |
Description
Specificity:
Goat Anti-Mouse 4-1BB recognizes Mouse 4-1BB. This antigen affinity purified polyclonal antibody was purified using a proprietary chromatographic technique that includes covalently immobilizing the antigen proteins or peptides to agarose based beads. This purification method enhances specificity, reduces nonspecific binding of extraneous IgG and provides you with the most reliable reagent available for your early discovery research.
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
Known Species Cross Reactivity: Ms+
Immunogen
NS0-Derived Recombinant Mouse 4-1BB Extracellular Domain
Formulation
This antigen affinity purified polyclonal 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
We suggest aseptically reconstituting this lyophilized, antigen affinity purified polyclonal antibody with 1 µl of sterile 1X PBS pH 7.2-7.3. This will result in a final antibody concentration of 0.1 mg/ml.
Endotoxin
<0.1 EU/µg as determined by the LAL method
Storage and Stability
The lyophilized antigen affinity purified polyclonal antibody can be stored desiccated at -20°C to -70°C for twelve months from date of receipt. 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 Donkey Anti-Goat IgG (H&L)-Horseradish Peroxidase (
Leinco Prod. No.: G505). The detection sensitivity of this indirect ELISA for Mouse 4-1BB is approximately 0.06 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 1 µg per 3.0-10.0 x 10
6 cells in a total staining volume of ≤200 µl followed by Donkey F(ab')
2 Anti-Goat IgG (H&L)-R-phycoerythrin (R-PE) (
Leinco Prod. No.: G641).
Western Blotting: To detect Mouse 4-1BB this polyclonal antibody can be used at a concentration of 0.1-0.2 µg/ml. This polyclonal antibody should be used in conjunction with compatible second-step reagents such as Donkey Anti-Goat IgG (H&L)-Horseradish Peroxidase (
Leinco Prod. No.: G505) and a chromogenic substrate such as TMB HRPO Membrane/IHC Substrate UltraSensitive™ One Component "Ready Use" (
Leinco Prod. No.: T343). The detection limit for Mouse 4-1BB is 1 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
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Products are for research use only. Not for use in diagnostic or therapeutic procedures.
