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Goat Anti-Human 4-1BB Receptor (4-1BBR)

Biotin

(TNFRSF9, CD137, T-cell Antigen ILA)

Polyclonal Antibody

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Prod. No.:	B392
Host:	Goat
Pkg. Size:	50 µg, 1 mg
Storage:	-20°C to -70°C Detailed storage instructions below.

Description

Specificity:

Goat Anti-Human 4-1BB Receptor recognizes Human 4-1BB Receptor. 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 Receptor, a member of the TNF superfamily of receptors, is mainly expressed on the surface of a variety of T cells, but also found in B cells, monocytes, and various transformed cell lines. 4-1BB Receptor binds to 4-1BBL to provide a co-stimulatory signal for T lymphocytes. Signaling by 4-1BB Receptor has been implicated in the antigen-presentation process and generation of cytotoxic T cells.

Known Species Cross Reactivity: Hu+

Immunogen

Purified Recombinant Human 4-1BB Receptor (>98%)

Formulation

This biotinylated 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 50 µg of bovine serum albumin per µg of antibody with no calcium, magnesium, or preservatives present.

Reconstitution

We suggest aseptically reconstituting this biotinylated, lyophilized, antigen affinity purified polyclonal antibody with 1 ml of sterile 20 mM Trizma base, 0.15 M sodium chloride (1X Tris Buffered Saline) pH 7.2-7.3, containing 0.1% w/v bovine serum albumin (BSA). This will result in a final biotinylated antibody concentration of 50 µg/ml.

Storage and Stability

The lyophilized, biotinylated antigen affinity purified polyclonal antibody can be stored desiccated at -20°C to -70°C for up to twelve months from date of receipt. The reconstituted bioin conjugate can be stored for at least four weeks at 2-8°C. For long-term storage of the reconstituted conjugate, 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

ELISA Det, FC, WB, , ICC

Flow Cytometry: It is recommended to use the indirect method for signal enhancement when enumerating cells expressing 4-1BB Receptor. A suggested method would be to stain cells expressing 4-1BB Receptor with 0.25 µg per 1 - 2.5 x 10⁵ cells in a total staining volume of ≤200 µl followed by UltraAvidin™- R-phycoerythrin (R-PE) (Leinco Prod. No.: A104).

Western Blotting: To detect Human 4-1BB Receptor this biotin conjugate can be used at a concentration of 0.1 - 0.2 µg/ml. This biotin conjugate should be used in conjunction with compatible second-step reagents such as UltraAvidin™- Horseradish Peroxidase (HRPO) (Leinco Prod. No.: A106) 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 Receptor is 0.5 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. A suitable Western blotting control is Recombinant Human 4-1BB Receptor (Leinco Prod. No.: F156).

ELISA Sandwich Assay: This antibody can be used as the detection antibody in a sandwich ELISA at a concentration of approximately 0.1 - 0.4 ng/ml when used in conjunction with Anti-Human 4-1BB Receptor (Antigen Aff. Pur.) (Leinco Prod. No.: B391) as the capture antibody at 0.4 µg/ml and an optimal second step reagent such as UltraAvidin™- Horseradish Peroxidase (HRPO) (Leinco Prod. No.: A106) for the detection of Human 4-1BB Receptor. The sensitivity of this sandwich ELISA is ~4 ng/ml.

Immunocytochemistry: This antibody will detect 4-1BB in cells. The working dilution is 2 - 15 μg/mL.

Each investigator should determine their own optimal working dilution for specific applications.

Country of Origin

USA