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PE Hamster Anti-Mouse CD54
PE Hamster Anti-Mouse CD54
Upregulation of CD54 expression on activated splenic B lymphocytes. Left panel: Naive BALB/c splenocytes were stained with PE-conjugated 3E2 mAb (open histogram) or unstained (filled histogram). Viable resting lymphocytes were gated according to scatter profile and exclusion of 7-AAD (BD Via-Probe™, Cat. No. 555816/555815). The mean fluorescence intensity of the stained lymphocytes is about 50 times greater than that of the negative-control lymphocytes. Right panel: 2-day LPS-activated BALB/c splenocytes were stained with PE-conjugated 3E2 mAb (open histogram) or unstained (filled histogram). Viable B-cell blasts were gated according to scatter profile and exclusion of 7-AAD. The mean fluorescence intensity of the stained blasts is about 180 times greater than that of the negative-control blasts. Flow cytometry was performed on a BD FACScan™ flow cytometry system.
Upregulation of CD54 expression on activated splenic B lymphocytes. Left panel: Naive BALB/c splenocytes were stained with PE-conjugated 3E2 mAb (open histogram) or unstained (filled histogram). Viable resting lymphocytes were gated according to scatter profile and exclusion of 7-AAD (BD Via-Probe™, Cat. No. 555816/555815). The mean fluorescence intensity of the stained lymphocytes is about 50 times greater than that of the negative-control lymphocytes. Right panel: 2-day LPS-activated BALB/c splenocytes were stained with PE-conjugated 3E2 mAb (open histogram) or unstained (filled histogram). Viable B-cell blasts were gated according to scatter profile and exclusion of 7-AAD. The mean fluorescence intensity of the stained blasts is about 180 times greater than that of the negative-control blasts. Flow cytometry was performed on a BD FACScan™ flow cytometry system.
Product Details
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BD Pharmingen™
ICAM-1; Icam1; Intercellular adhesion molecule 1; Ly-47; MALA-2; MyD10
Mouse (QC Testing)
Armenian Hamster IgG1, κ
Not reported
Flow cytometry (Routinely Tested)
0.2 mg/ml
15894
AB_394735
Aqueous buffered solution containing ≤0.09% sodium azide.
RUO


Preparation And Storage

The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated with R-PE under optimum conditions, and unconjugated antibody and free PE were removed. Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.

Recommended Assay Procedures

This antibody conjugate is compatible with intracellular staining protocols using the BD Cytofix/Cytoperm™ Kit (Cat. No. 554714).

Product Notices

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  2. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  3. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  4. Although hamster immunoglobulin isotypes have not been well defined, BD Biosciences Pharmingen has grouped Armenian and Syrian hamster IgG monoclonal antibodies according to their reactivity with a panel of mouse anti-hamster IgG mAbs. A table of the hamster IgG groups, Reactivity of Mouse Anti-Hamster Ig mAbs, may be viewed at http://www.bdbiosciences.com/documents/hamster_chart_11x17.pdf.
  5. 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.
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Antibody Details
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3E2

The 3E2 monoclonal antibody specifically binds to CD54 (ICAM-1), a 95-kDa member of the Ig superfamily found on lymphocytes, vascular endothelium, high endothelial venules, epithelial cells, macrophages, and dendritic cells. ICAM-1 is a ligand for LFA1 (CD11a/CD18) and Mac-1 (CD11b/CD18). Its expression is upregulated upon stimulation by inflammatory mediators such as cytokines and LPS. Studies with mouse Icam1-transfected antigen-presenting cells, with CD54-blocking antibodies, and in CD54-deficient mice indicate that CD54 participates in inflammatory reactions and antigen-specific immune responses. In addition, there is evidence that CD54 is a receptor involved in MHC-non-restricted responses to weakly immunogenic tumor cells. The 3E2 antibody has been reported to block in vitro and in vivo intracellular adhesion events involved in immune responses.

This antibody is routinely tested by flow cytometric analysis. Other applications were tested at BD Biosciences Pharmingen during antibody development only or reported in the literature.

553253 Rev. 17
Format Details
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PE
R-Phycoerythrin (PE), is part of the BD family of Phycobiliprotein dyes. This fluorochrome is a multimeric fluorescent phycobiliprotein with excitation maximum (Ex Max) of 496 nm and 566 nm and an emission maximum (Em Max) at 576 nm. PE is designed to be excited by the Blue (488 nm), Green (532 nm) and Yellow-Green (561 nm) lasers and detected using an optical filter centered near 575 nm (e.g., a 575/26-nm bandpass filter). As PE is excited by multiple lasers, this can result in cross-laser excitation and fluorescence spillover on instruments with various combinations of Blue, Green, and Yellow-Green lasers. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
altImg
PE
Yellow-Green 488 nm, 532 nm, 561 nm
496 nm, 566 nm
576 nm
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Citations & References
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Development References (13)

  1. Gonzalo JA, Martinez C, Springer TA, Gutierrez-Ramos JC. ICAM-1 is required for T cell proliferation but not for anergy or apoptosis induced by Staphylococcus aureus enterotoxin B in vivo. Int Immunol. 1995; 7(10):1691-1698. (Biology). View Reference
  2. Isobe M, Yagita H, Okumura K, Ihara A. Specific acceptance of cardiac allograft after treatment with antibodies to ICAM-1 and LFA-1. Science. 1992; 255(5048):1125-1127. (Biology). View Reference
  3. Kelly KJ, Williams WW Jr, Colvin RB, et al. Intercellular adhesion molecule-1-deficient mice are protected against ischemic renal injury. J Clin Invest. 1996; 97(4):1056-1063. (Biology). View Reference
  4. Masten BJ, Yates JL, Pollard Koga AM, Lipscomb MF. Characterization of accessory molecules in murine lung dendritic cell function: roles for CD80, CD86, CD54, and CD40L. Am J Respir Cell Mol Biol. 1997; 16(3):335-342. (Clone-specific). View Reference
  5. Nishio M, Podack ER. Rapid induction of tumor necrosis factor cytotoxicity in naive splenic T cells by simultaneous CD80 (B7.1) and CD54 (ICAM-1) co-stimulation. Eur J Immunol. 1996; 26(9):2160-2164. (Biology). View Reference
  6. Nishio M, Spielman J, Lee RK, Nelson DL, Podack ER. CD80 (B7.1) and CD54 (intracellular adhesion molecule-1) induce target cell susceptibility to promiscuous cytotoxic T cell lysis. J Immunol. 1996; 157(10):4347-4353. (Biology). View Reference
  7. Scheynius A, Camp RL, Pure E. Reduced contact sensitivity reactions in mice treated with monoclonal antibodies to leukocyte function-associated molecule-1 and intercellular adhesion molecule-1. J Immunol. 1993; 150(2):655-663. (Clone-specific). View Reference
  8. Scheynius A, Camp RL, Pure E. Unresponsiveness to 2,4-dinitro-1-fluoro-benzene after treatment with monoclonal antibodies to leukocyte function-associated molecule-1 and intercellular adhesion molecule-1 during sensitization. J Immunol. 1996; 154(5):1804-1809. (Biology). View Reference
  9. Siu G, Hedrick SM, Brian AA. Isolation of the murine intercellular adhesion molecule 1 (ICAM-1) gene. ICAM-1 enhances antigen-specific T cell activation. J Immunol. 1989; 143(11):3813-3820. (Biology). View Reference
  10. Soriano SG, Lipton SA, Wang YF, et al. Intercellular adhesion molecule-1-deficient mice are less susceptible to cerebral ischemia-reperfusion injury. Ann Neurol. 1996; 39(5):618-624. (Biology). View Reference
  11. Springer TA. Adhesion receptors of the immune system. Nature. 1990; 346(6283):425-434. (Clone-specific). View Reference
  12. Springer TA. Traffic signals for lymphocyte recirculation and leukocyte emigration: the multistep paradigm. Cell. 1994; 76(2):301-314. (Biology). View Reference
  13. Xu H, Gonzalo JA, St Pierre Y, et al. Leukocytosis and resistance to septic shock in intercellular adhesion molecule 1-deficient mice. J Exp Med. 1994; 180(1):95-109. (Biology). View Reference
View All (13) View Less
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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.

Refer to manufacturer's instructions for use and related User Manuals and Technical Data Sheets before using this product 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.