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Anti-Human IL-12

产品编号:I-351

产品描述:Anti-Human IL-12

反应种属:human

实验方法:ELISA, N, WB, ICC

标记:

规格:500ug

供应商:leinco

价格(RMB):4650.00

订购:订购

说明书:

Anti-Human Interleukin-12 (IL-12)
Purified No Carrier Protein
(NKSF, TSF, Maturation Factor, Cytotoxic Lymphocyte Maturation Factor (CLMF), p35)
Monoclonal Antibody
Prod. No.: I-351
Clone:     24910
Isotype:   Mouse IgG1
Pkg. Size: 500ug
Storage:   -20°C to -70°C

Description
Specificity:
Mouse Anti-Human Interleukin-12 (IL-12) (Clone 24910) recognizes an epitope on Human IL-12. This monoclonal antibody was purified using multi-step affinity chromatography methods such as Protein A or G depending on the species and isotype.

Background:

Interleukin 12 (IL-12, NK cell stimulatory factor, cytotoxic lymphocyte maturation factor) is a heterodimeric cytokine that is naturally produced by dendritic cells1 , macrophages and human B-lymphoblastoid cells (NC-37) in response to antigenic stimulation. IL-12 is involved in the differentiation of naive T cells into Th1 cells, which is important in resistance against pathogens. It is known as a T cell stimulating factor, which can stimulate the growth and function of T cells. It stimulates the production of IFN-γ and TNF-α from T and natural killer (NK) cells, and reduces IL-4 mediated suppression of IFN-γ. IL-12 also has anti-angiogenic activity, which can block the formation of new blood vessels. IL-12 binds to the IL-12 receptor and upon binding, IL-12R-β2 becomes tyrosine phosphorylated and provides binding sites for kinases, Tyk2 and Jak2. These are important in activating critical transcription factor proteins such as STAT4 which are implicated in IL-12 signaling in T cells and NK cells.2 IL-12 contributes to the antimycobacterial immune response by enhancing production of interferon-gamma, facilitating development of Th1 cells and augmenting cytotoxicity of antigen-specific T cells and natural killer cells.3

Known Species Cross Reactivity: Hu+
Host Species
Mouse
Immunogen
Purified Recombinant Sf 21-derived rhIL-12 (>98%)
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 should be aseptically reconstituted with sterile PBS to a final volume of 1.0 ml. This would result in a final antibody concentation of 0.5 mg/ml. Higher concentrations may be possible.
Endotoxin
<0.1 EU/µg
Storage and Stability
The lyophilized antibody should be stored desiccated at -20°C to -70°C. 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, N, WB, ICC
Indirect ELISA: This antibody can be used in an indirect detection ELISA at 5-25 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 IL-12 is approximately 3 ng/well.
Neutralization: Human IL-12 stimulates the 3H-thymidine incorporation by PHA activated human peripheral blood mononuclear cells in a dose-dependent manner1. The ED50 for this effect is typically 0.05 - 0.2 ng/mL. This antibody is useful for neutralization of Human IL-12 bioactivity. The antibody dose required to neutralize 50% (ND50) of the biological activity of Human IL-12 (at 1.0 ng/ml) is 0.3 - 0.9 µg/ml. The bioassay was perfromed using PHA-activated peripheral blood mononuclear cell (PBMC) proliferation assay.
Western Blotting: To detect Human IL-12 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 IL-12 is 20 ng/lane under non-reducing conditions. Use of this antibody under reducing conditions is not recommended. The sensitivity of detection may increase up to 50 fold when a chemiluminescent substrate is used. A suitable Western blotting control is Recombinant Human IL-12 (expressed in Sf 21 insect cells) (Leinco Prod. No.: I-535).
Immunocytochemistry: This antibody can be used at 5 - 25 μg/mL with the appropriate secondary reagent such as Goat F(ab')2 Anti-Mouse IgG (H&L)-Horseradish Peroxidase (Leinco Prod. No.: M103) to detect human IL-12 in cultured cells or tissue sections using Optional Metal Enhanced DAB Substrate Kit (Leinco Prod. No.: K107) or 3, 3'-Diaminobenzidine Tetrahydrochloride (DAB) Substrate Kit (Leinco Prod. No.: D100).

References
1. Kapsenberg ML et al. (1997) J. Immunol. 159: 28
2. Jerome Ritz. et al. Blood 95: 3190
3. Barnes PF. et al. J. Clin. Invest. 93: 1733

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Recombinant Human IL-12 (expressed in Sf 21 insect cells) (Leinco Prod. No.: I-535)
Goat F(ab')2 Anti-Mouse IgG (H&L)-Horseradish Peroxidase (Leinco Prod. No.: M103)
Optional Metal Enhanced DAB Substrate Kit (Leinco Prod. No.: K107)
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Products are for research use only. Not for use in diagnostic or therapeutic procedures