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      Purified Hamster Anti-Mouse CD54
      01541D_553250_image2.png

      Upregulation of CD54 expression on activated splenic B lymphocytes. Left panel: Naive BALB/c splenocytes were stained with purified 3E2 mAb (open histogram) followed by FITC-conjugated anti-hamster IgG cocktail (Cat. No. 554011, filled and open histograms). 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 7.5 times greater than that of the negative-control lymphocytes. Right panel: 2-day LPS-activated BALB/c splenocytes were stained with purified 3E2 mAb (open histogram) followed by FITC-conjugated anti-hamster IgG (filled and open histograms). 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 15 times greater than that of the negative-control blasts. Flow cytometry was performed on a BD FACScan™ flow cytometry system.

      553250Image1.png

      Upregulation of CD54 expression on activated splenic B lymphocytes. Left panel: Naive BALB/c splenocytes were stained with purified 3E2 mAb (open histogram) followed by FITC-conjugated anti-hamster IgG cocktail (Cat. No. 554011, filled and open histograms). 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 7.5 times greater than that of the negative-control lymphocytes. Right panel: 2-day LPS-activated BALB/c splenocytes were stained with purified 3E2 mAb (open histogram) followed by FITC-conjugated anti-hamster IgG (filled and open histograms). 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 15 times greater than that of the negative-control blasts. Flow cytometry was performed on a BD FACScan™ flow cytometry system.

      01541D_553250_image2.png 553250Image1.png Blue-Cap-White-Label.jpg

      Upregulation of CD54 expression on activated splenic B lymphocytes. Left panel: Naive BALB/c splenocytes were stained with purified 3E2 mAb (open histogram) followed by FITC-conjugated anti-hamster IgG cocktail (Cat. No. 554011, filled and open histograms). 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 7.5 times greater than that of the negative-control lymphocytes. Right panel: 2-day LPS-activated BALB/c splenocytes were stained with purified 3E2 mAb (open histogram) followed by FITC-conjugated anti-hamster IgG (filled and open histograms). 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 15 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 purified 3E2 mAb (open histogram) followed by FITC-conjugated anti-hamster IgG cocktail (Cat. No. 554011, filled and open histograms). 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 7.5 times greater than that of the negative-control lymphocytes. Right panel: 2-day LPS-activated BALB/c splenocytes were stained with purified 3E2 mAb (open histogram) followed by FITC-conjugated anti-hamster IgG (filled and open histograms). 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 15 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), Blocking, Immunohistochemistry-frozen (Reported)
      0.5 mg/ml
      AB_394732
      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. Store undiluted at 4°C.
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      Recommended Assay Procedures

      For IHC, we recommend the use of purified 3E2 mAb in our special formulation for immunohistochemistry, Cat. No. 550287.

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      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. 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.
      4. 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.
      5. Sodium azide is a reversible inhibitor of oxidative metabolism; therefore, antibody preparations containing this preservative agent must not be used in cell cultures nor injected into animals. Sodium azide may be removed by washing stained cells or plate-bound antibody or dialyzing soluble antibody in sodium azide-free buffer. Since endotoxin may also affect the results of functional studies, we recommend the NA/LE (No Azide/Low Endotoxin) antibody format, if available, for in vitro and in vivo use.
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      553250 Rev. 15
      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.

      553250 Rev. 15
      Format Details
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      Purified
      Tissue culture supernatant is purified by either protein A/G or affinity purification methods. Both methods yield antibody in solution that is free of most other soluble proteins, lipids, etc. This format provides pure antibody that is suitable for a number of downstream applications including: secondary labeling for flow cytometry or microscopy, ELISA, Western blot, etc.
      Purified
      553250 Rev.15
      Citations & References
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      View product citations for antibody "553250" on CiteAb

      Development References (14)

      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: Blocking). 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. Ritter DM, McKerrow JH. Intercellular adhesion molecule 1 is the major adhesion molecule expressed during schistosome granuloma formation. Infect Immun. 1996; 64(11):4706-4713. (Clone-specific: Immunohistochemistry). View Reference
      8. 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: Blocking, Immunohistochemistry). View Reference
      9. 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
      10. 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
      11. 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
      12. Springer TA. Adhesion receptors of the immune system. Nature. 1990; 346(6283):425-434. (Clone-specific: Blocking). View Reference
      13. Springer TA. Traffic signals for lymphocyte recirculation and leukocyte emigration: the multistep paradigm. Cell. 1994; 76(2):301-314. (Biology). View Reference
      14. 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 (14) View Less
      553250 Rev. 15

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