PDF《Monoclonal》

Monoclonal Anti-mouse Limitin Antibody FOR RESEARCH USE ONLY.

NOT FOR USE IN HUMANS. R&D Systems, Inc. 1-800-343-7475 11/26/07 ORDERING INFORMATION Catalog Number: MAB5971 Clone:

183707 Lot Number: ZTP01 Size:

500 μg Formulation: 0.2 μm filtered solution in PBS with 5% trehalose Storage: -20° C Reconstitution: sterile PBS Specificity: mouse Limitin Immunogen: NS0-derived rmLimitin Ig class: rat IgG2A Recommended Applications: Neutralization of bioactivity ELISA capture Background Limitin, also known as IFN-zeta, is a

20 kDa glycoprotein that is secreted by T cells and bronchial and salivary duct epithelial cells. It inhibits B-lineage cell development but has little effect on myelopoiesis or erythropoiesis. Limitin binds to IFN-α/β receptors and has potent antiviral activity. Mouse Limitin shares 32%, 26%, and 30% amino acid sequence identity with mouse IFN-α, mouse IFN-β, and human IFN-ω, respectively. Preparation This antibody was produced from a hybridoma resulting from the fusion of a mouse myeloma with B cells obtained from a rat immunized with purified, NS0-derived, recombinant mouse Limitin (rmLimitin;

aa

22 - 182;

Accession # NP_001079001). The IgG fraction of the tissue culture supernatant was purified by Protein G affinity chromatography. Formulation Lyophilized from a 0.2 μm filtered solution in phosphate-buffered saline (PBS) with 5% trehalose. Endotoxin Level < 0.1 EU per

1 μg of the antibody as determined by the LAL method. Reconstitution Reconstitute with sterile PBS. If

1 mL of PBS is used, the antibody concentration will be

500 μg/mL. Storage Lyophilized samples are stable for twelve months from date of receipt when stored at -20° C to -70° C. Upon reconstitution, the antibody can be stored at 2° - 8° C for

1 month without detectable loss of activity. Reconstituted antibody can also be aliquotted and stored frozen at -20° C to -70° C in a manual defrost freezer for six months without detectable loss of activity. Avoid repeated freeze-thaw cycles. Specificity This antibody detects rmLimitin in direct ELISAs and Western blots. In direct ELISAs, this antibody shows no cross-reactivity with rmIFNγ. Applications Neutralization of mouse LImitin bioactivity - The exact concentration of antibody required to neutralize mouse Limitin activity is dependent on the cytokine concentration, cell type, growth conditions and the type of activity studied. To provide a guideline, R&D Systems has determined the neutralization dose for this antibody under a specific set of conditions. The Neutralization Dose50 (ND50) for this antibody is defined as that concentration of antibody required to yield one-half maximal inhibition of the cytokine activity on a responsive cell line, when that cytokine is present at a concentration just high enough to elicit a maximum response. The ND50 for this lot of anti-mouse Limitin antibody was determined to be approximately 0.02 - 0.1 μg/mL in the presence of 0.03 ng/mL of rmLimitin, using the L929 cell line. The specific conditions are described in the figure legends. ELISA capture - This product can be used as a capture reagent in a mouse Limitin sandwich immunoassay in combination with biotinylated mouse Limitin detection antibody (Cat. # BAF597) and recombinant mouse Limitin (Cat. # 1535-LM) as the standard. The suggested coating concentration range is

2 -

8 μg/mL and should be titrated to determine the optimal concentration. A general protocol is provided at www.RnDSystems.com/go/MAPELISA. Optimal dilutions should be determined by each laboratory for each application.