Multicolor flow cytometric analysis of CD100 expression on human peripheral blood cells. Human peripheral blood was treated with BD PharmLyse™ Lysing Buffer to lyse erythrocytes. The leucocytes were washed and stained with Alexa Fluor 488 Mouse Anti-Human CD4 (Cat. No. 557695; Upper Panels) and BD Horizon™ V450 Mouse Anti-Human CD19 (Cat. No. 560353; Lower Panels) antibodies, and with either Alexa Fluor® 647 Mouse IgG1, κ Isotype Control (Cat No. 557714; Left Panels) or Alexa Fluor® 647 Mouse Anti-Human CD100 (Cat. No. 564873; Right Panels). The two-color flow cytometric contour plots showing the correlated expression of CD4 or CD19 versus CD100 (or Ig Isotype control staining) were derived ffrom gated events with the forward and side light-scatter characteristics of viable lymphocytes. Flow cytometric analysis was performed using a BD LSRFortessa™ Flow Cytometer System.
Multicolor flow cytometric analysis of CD100 expression on human peripheral blood cells. Human peripheral blood was treated with BD PharmLyse™ Lysing Buffer to lyse erythrocytes. The leucocytes were washed and stained with Alexa Fluor 488 Mouse Anti-Human CD4 (Cat. No. 557695; Upper Panels) and BD Horizon™ V450 Mouse Anti-Human CD19 (Cat. No. 560353; Lower Panels) antibodies, and with either Alexa Fluor® 647 Mouse IgG1, κ Isotype Control (Cat No. 557714; Left Panels) or Alexa Fluor® 647 Mouse Anti-Human CD100 (Cat. No. 564873; Right Panels). The two-color flow cytometric contour plots showing the correlated expression of CD4 or CD19 versus CD100 (or Ig Isotype control staining) were derived ffrom gated events with the forward and side light-scatter characteristics of viable lymphocytes. Flow cytometric analysis was performed using a BD LSRFortessa™ Flow Cytometer System.
Product Details
BD Pharmingen™
Semaphorin-4D ; SEMA4D; SEM4D; SEMAJ; GR3; C9orf164
Human (QC Testing)
Mouse IgG1
Flow cytometry (Routinely Tested)
5 µl
V BP383, S233
AB_2738993
Aqueous buffered solution containing BSA and ≤0.09% sodium azide.
RUO
Preparation And Storage
Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated to Alexa Fluor® 647 under optimum conditions, and unreacted Alexa Fluor® 647 was removed.
Product Notices
- This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test).
- An isotype control should be used at the same concentration as the antibody of interest.
- Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
- Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
- The Alexa Fluor®, Pacific Blue™, and Cascade Blue® dye antibody conjugates in this product are sold under license from Molecular Probes, Inc. for research use only, excluding use in combination with microarrays, or as analyte specific reagents. The Alexa Fluor® dyes (except for Alexa Fluor® 430), Pacific Blue™ dye, and Cascade Blue® dye are covered by pending and issued patents.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- 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.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
Companion Products
Stain Buffer (FBS) RUO
Size
500 mL
Cat No.
554656
Stain Buffer (BSA) RUO
Size
500 mL
Cat No.
554657
Lysing Buffer RUO
Size
100 mL
Cat No.
555899
Alexa Fluor® 647 Mouse IgG1 κ Isotype Control RUO
Size
100 Tests
Cat No.
557714
Alexa Fluor®488 Mouse Anti-Human CD4 RUO
Size
100 Tests
Cat No.
557695
V450 Mouse Anti-Human CD19 RUO
Size
120 Tests
Cat No.
560353
564873 Rev. 1
Antibody Details
A8
The A8 monoclonal antibody specifically binds to CD100 which is also known as Semaphorin-4D (SEMA4D). CD100 is a 150 kDa type I transmembrane glycoprotein that belongs to the semaphorin family. CD100 is widely expressed in the nervous and immune systems. Although it is expressed on T cells, its expression is significantly upregulated on activated T cells. It is also expressed on NK cells, B cells, and monocytes. CD100 is an adhesion molecule that functions as a signaling molecule which plays a role in lymphocyte activation. CD100 interaction with CD72 enhances B cell signaling and ultimately promotes B cell activation and survival.
564873 Rev. 1
Format Details
Alexa Fluor™ 647
Alexa Fluor™ 647 Dye is part of the BD red family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 653-nm and an emission maximum (Em Max) at 669-nm. Alexa Fluor 647 is designed to be excited by the Red laser (627-640 nm) and detected using an optical filter centered near 520-nm (e.g., a 660/20 nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
Alexa Fluor™ 647
Red 627-640 nm
653 nm
669 nm
564873 Rev.1
Citations & References
View product citations for antibody "564873" on CiteAb
Development References (5)
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Hall KT, Boumsell L, Schultze JL, et al. Human CD100, a novel leukocyte semaphorin that promotes B-cell aggregation and differentiation. Proc Natl Acad Sci U S A. 1996; 93(21):11780-11785. (Clone-specific: Flow cytometry, Fluorescence activated cell sorting). View Reference
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Kikutani H, Kumanogoh A. Semaphorins in interactions between T cells and antigen-presenting cells. Nat Rev Immunol. 2003; 3(2):159-167. (Biology). View Reference
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Kumanogoh A, Watanabe C, Lee I, et al. Identification of CD72 as a lymphocyte receptor for the class IV semaphorin CD100: a novel mechanism for regulating B cell signaling. Immunity. 2000; 13(5):621-631. (Biology). View Reference
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Li DH, Tung JW, Tarner IH, et al. CD72 down-modulates BCR-induced signal transduction and diminishes survival in primary mature B lymphocytes. J Immunol. 2006; 176(9):5321-5328. (Biology). View Reference
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Mou P, Zeng Z, Li Q, et al. Identification of a calmodulin-binding domain in Sema4D that regulates its exodomain shedding in platelets. Blood. 2013; 121(20):4221-4230. (Clone-specific: Flow cytometry). View Reference
564873 Rev. 1
Please refer to Support Documents for Quality Certificates
Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described
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.