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Purified NA/LE Rat Anti-Mouse CD275
Purified NA/LE Rat Anti-Mouse CD275
Two-color flow cytometric analysis of CD275 expression on mouse splenocytes. Mouse splenic leucocytes were stained with either Purified Rat IgG2a, κ Isotype Control (Cat. No. 553927; Left Plot) or Purified NA/LE Rat Anti-Mouse CD275 antibody (Cat. No. 566746; Right Plot) at 0.5 µg/test. The cells were washed and stained with FITC Goat Anti-Rat Ig (Cat. No. 554016). The cells were washed, preincubated with Rat IgG Isotype Control (ThermoFisher Scientific; Cat No: 10700) at 15 µg/test, and then stained with Alexa Fluor® 647 Rat Anti-Mouse CD45R/B220 (Cat. No. 557683). The pseudocolor dot plot showing the correlated expression of CD275 (or Ig Isotype control staining) versus CD45R/B220 was derived from gated events with the forward and side-light scatter characteristics of viable lymphocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
Two-color flow cytometric analysis of CD275 expression on mouse splenocytes. Mouse splenic leucocytes were stained with either Purified Rat IgG2a, κ Isotype Control (Cat. No. 553927; Left Plot) or Purified NA/LE Rat Anti-Mouse CD275 antibody (Cat. No. 566746; Right Plot) at 0.5 µg/test. The cells were washed and stained with FITC Goat Anti-Rat Ig (Cat. No. 554016). The cells were washed, preincubated with Rat IgG Isotype Control (ThermoFisher Scientific; Cat No: 10700) at 15 µg/test, and then stained with Alexa Fluor® 647 Rat Anti-Mouse CD45R/B220 (Cat. No. 557683). The pseudocolor dot plot showing the correlated expression of CD275 (or Ig Isotype control staining) versus CD45R/B220 was derived from gated events with the forward and side-light scatter characteristics of viable lymphocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
Product Details
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BD Pharmingen™
Icoslg; ICOS-L; B7-H2; B7RP-1; B7h; ICOS ligand; Icosl; LICOS; Ly115l
Mouse (QC Testing)
Rat SD, also known as Sprague-Dawley (outbred) IgG2a, κ
Mouse B7RP-1 Transfected Cell Line
Flow cytometry (Routinely Tested)
1.0 mg/ml
AB_2869844
No azide/low endotoxin: Aqueous buffered solution containing no preservative, 0.2µm sterile filtered. Endotoxin level is ≤0.1 EU/µg (≤0.01 ng/µg) of protein as determined by the LAL assay.
RUO


Preparation And Storage

Store undiluted at 4°C. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. This preparation contains no preservatives, thus it should be handled under aseptic conditions.

Product Notices

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  2. An isotype control should be used at the same concentration as the antibody of interest.
  3. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
566746 Rev. 1
Antibody Details
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HK5.3

The HK5.3 monoclonal antibody specifically binds to CD275 which is also known as B7 homolog 2 (B7-H2 or B7h2), B7-related protein 1 (B7RP-1), B7-like protein Gl50, ICOS Ligand (ICOS-L), LICOS, or Ly115l. CD275 is a type I transmembrane glycoprotein that is encoded by Icoslg (icos ligand) which belongs to the B7 family of costimulatory molecules within the Ig gene superfamily. Two isoforms of mouse CD275 have been described. CD275 is variably expressed on antigen-presenting cells including B cells, macrophages, and dendritic cells. CD275 serves as a costimulatory molecule that binds to and signals through CD278 (ICOS) which is expressed on activated T cells to increase their proliferation, production of cytokines, and provision of help for B cells. CD275 expression is upregulated by TNF and can help mediate local tissue responses to inflammatory conditions. The HK5.3 antibody reportedly blocks the binding of CD275/ICOS Ligand to its receptor, CD278 (ICOS), both in vitro and in vivo, and thus inhibits its biological functions.

                

566746 Rev. 1
Format Details
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NA/LE
NA/LE refers to the culture and purification methods and buffer used to produce purified antibodies with no azide and low endotoxin: Aqueous buffered solution containing no preservative, 0.2µm sterile filtered. Endotoxin level is ≤0.01 EU/µg (≤0.001 ng/µg) of protein as determined by the LAL assay.NA/LE are perfectly suited to be used in culture or in vivo (for nonhuman studies) for functional assays — blocking, neutralizing, activation or depletion — where the presence of azide may damage cells or exogenous endotoxin may signal or activate cells.
NA/LE
566746 Rev.1
Citations & References
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Development References (10)

  1. Carreno BM, Collins M. The B7 family of ligands and its receptors: New pathways for costimulation and inhibition of immune responses. Annu Rev Immunol. 2002; 20:29-53. (Biology). View Reference
  2. Crotty S, Choi YS, Kageyama R, et al. ICOS receptor instructs T follicular helper cell versus effector cell differentiation via induction of the transcriptional repressor Bcl6. Immunity. 2011; 34:1-15. (Clone-specific: Inhibition, In vivo exacerbation). View Reference
  3. Frauwirth KA, Thompson CB. Activation and inhibition of lymphocytes by costimulation. J Clin Invest. 2002; 109(3):295-299. (Biology). View Reference
  4. Iwai H, Kozono Y, Hirose S, et al. Amelioration of collagen-induced arthritis by blockade of inducible costimulator-B7 homologous protein costimulation. J Immunol. 2000; 169(8):4332-4339. (Immunogen: Blocking, Flow cytometry, Functional assay, Inhibition, In vivo exacerbation). View Reference
  5. Kohyama M, Sugahara D, Sugiyama S, Yagita H, Okumura K, Hozumi N. Inducible costimulator-dependent IL-10 production by regulatory T cells specific for self-antigen. Proc Natl Acad Sci U S A. 2004; 101(12):4192-4197. (Clone-specific: Blocking, Functional assay, Inhibition). View Reference
  6. Ling V, Wu PW, Finnerty HF, et al. Cutting edge: identification of GL50, a novel B7-like protein that functionally binds to ICOS receptor. J Immunol. 2000; 164(4):1653-1657. (Biology). View Reference
  7. Liu X, Bai XF, Wen J, et al. B7H costimulates clonal expansion of, and cognate destruction of tumor cells by, CD8(+) T lymphocytes in vivo. J Exp Med. 2001; 194(9):1339-1348. (Biology). View Reference
  8. Ozkaynak E, Gao W, Shemmeri N, et al. Importance of ICOS-B7RP-1 costimulation in acute and chronic allograft rejection. Nat Immunol. 2001; 2(7):591-596. (Biology). View Reference
  9. Swallow MM, Wallin JJ, Sha WC. B7h, a novel costimulatory homolog of B7.1 and B7.2, is induced by TNFalpha. Immunity. 1999; 11(4):423-432. (Biology). View Reference
  10. Yoshinaga SK, Whoriskey JS, Khare SD, et al. T-cell co-stimulation through B7RP-1 and ICOS. Nature. 1999; 402(6763):827-832. (Biology). View Reference
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566746 Rev. 1

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For Research Use Only. Not for use in diagnostic or therapeutic procedures.