BD Pharmingen™ Purified Mouse Anti-Human CD130
クローン AM64 (RUO)
Preparation and Storage
推奨アッセイ手順
For flow cytometric applications, a three step labelling procedure is recommended for amplifying signal. Suggested protocol for 3-step staining using Lysed Whole Blood method:
1. Incubate 100 µl whole blood with primary (unconjugated) antibody for 20-30 minutes in the dark at room temperature.
2. Add 2 mls of 1X Pharm Lyse (10X Pharm Lyse, Cat. No. 555899) and incubate for 10-15 minutes in the dark. Centrifuge and aspirate.
3. Wash once with PBS/0.1% sodium azide/1% heat-inactivated fetal bovine serum (PBS-FBS). Centrifuge and aspirate.
4. Add biotinylated goat anti-mouse Ig's and incubate for 20-30 minutes in the dark at room temperature.
5. Wash once with PBS-FBS. Centrifuge and aspirate.
6. Add SAV-PE (Cat. No. 554061) and incubate for 20-30 minutes in the dark at room temperature.
7. Wash once with PBS-FBS. Centrifuge and aspirate. Resuspend in 0.5 ml of PBS-FBS and analyze by flow cytometry.
Product Notices
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
The AM64 monoclonal antibody specifically binds to CD130 which is also known as gp130 (Membrane glycoprotein 130), IL6ST (Interleukin-6 signal transducer), and IL6RB/IL-6Rβ (Interleukin-6 Receptor beta). CD130 is a 130 kDa type 1 membrane glycoprotein that belongs to the hemopoietic cytokine receptor superfamily. CD130 serves as a common signaling receptor for several cytokine receptor complexes including those that mediate the biological effects of IL-6, IL-11, IL-27, Cardiotrophin-1 (CT-1), Ciliary neurotrophic factor (CNTF), Leukemia inhibitory factor (LIF), and Oncostatin M (OSM). CD130 is widely expressed on a variety of cell types including T cells, activated B cells, monocytes, plasma cells, and endothelia. The AM64 antibody can be used in flow cytometry, immunoprecipitation, western blot, and immunohistochemistry on frozen sections.
Development References (3)
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Hibi M, Murakami M, Saito M, Hirano T, Taga T, Kishimoto T. Molecular cloning and expression of an IL-6 signal transducer, gp130. Cell. 1990; 63(6):1149-1157. (Biology). View Reference
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Pietzko D, Zohlnhöfer D, Graeve L, et al. The hepatic interleukin-6 receptor. Studies on its structure and regulation by phorbol 12-myristate 13-acetate-dexamethasone. J Biol Chem. 1993; 268(6):4250-4258. (Biology). View Reference
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Schlossman SF. Stuart F. Schlossman .. et al., ed. Leucocyte typing V : white cell differentiation antigens : proceedings of the fifth international workshop and conference held in Boston, USA, 3-7 November, 1993. Oxford: Oxford University Press; 1995.
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Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims. Comparisons are not made against non-BD technologies, unless otherwise noted.
For Research Use Only. Not for use in diagnostic or therapeutic procedures.