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R718 Hamster Anti-Mouse CD48
R718 Hamster Anti-Mouse CD48
Flow cytometric analysis of CD48 on mouse splenocytes. Mouse splenic leukocytes from a Balb/c mouse were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142). The cells were stained with no antibody [for a Fluorescence-Minus-One (FMO) control; dashed line histogram] or stained with BD Horizon™ R718 Hamster Anti-Mouse CD48 antibody (Cat. No. 567301; solid line histogram) at 0.5 µg/test. DAPI (4',6-Diamidino-2-Phenylindole, Dihydrochloride) Solution (Cat. No. 564907) was added to cells right before analysis. Fluorescence histograms showing CD48 expression (or FMO control) were derived from gated events with the forward and side light-scattering characteristics of viable (DAPI-negative) splenocytes. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometry System. Data shown on this Technical Data Sheet are not lot specific.
Flow cytometric analysis of CD48 on mouse splenocytes. Mouse splenic leukocytes from a Balb/c mouse were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142). The cells were stained with no antibody [for a Fluorescence-Minus-One (FMO) control; dashed line histogram] or stained with BD Horizon™ R718 Hamster Anti-Mouse CD48 antibody (Cat. No. 567301; solid line histogram) at 0.5 µg/test. DAPI (4',6-Diamidino-2-Phenylindole, Dihydrochloride) Solution (Cat. No. 564907) was added to cells right before analysis. Fluorescence histograms showing CD48 expression (or FMO control) were derived from gated events with the forward and side light-scattering characteristics of viable (DAPI-negative) splenocytes. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometry System. Data shown on this Technical Data Sheet are not lot specific.
Product Details
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BD Horizon™
BLAST; BLAST-1; BCM1; HM48-1; MEM-102; Sgp-60; SLAMF2
Mouse (QC Testing)
Armenian Hamster IgG1, λ3
Mouse T lymphoma MBL-2
Flow cytometry (Routinely Tested)
0.2 mg/ml
12506
Aqueous buffered solution containing ≤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 the dye under optimum conditions and unreacted dye was removed.

Recommended Assay Procedures

BD® CompBeads can be used as surrogates to assess fluorescence spillover (Compensation). When fluorochrome conjugated antibodies are bound to BD® CompBeads, they have spectral properties very similar to cells. However, for some fluorochromes there can be small differences in spectral emissions compared to cells, resulting in spillover values that differ when compared to biological controls. It is strongly recommended that when using a reagent for the first time, users compare the spillover on cells and BD CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.

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. 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.
  4. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  5. This product is provided under an Agreement between BIOTIUM and BD Biosciences. This product, and only in the amount purchased by buyer, may be used solely for buyer’s own internal research, in a manner consistent with the accompanying product literature. No other right to use, sell or otherwise transfer (a) this product, or (b) its components is hereby granted expressly, by implication or by estoppel. This product is for research use only. Diagnostic uses require a separate license from Biotium, Inc. For information on purchasing a license to this product including for purposes other than research, contact Biotium, Inc., 3159 Corporate Place, Hayward, CA 94545, Tel: (510) 265-1027. Fax: (510) 265-1352. Email: btinfo@biotium.com.
  6. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  7. Alexa Fluor™ is a trademark of Life Technologies Corporation.
  8. 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.
567301 Rev. 1
Antibody Details
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HM48-1

The HM48-1 monoclonal antibody specifically binds to CD48 (previously known as BCM1 in mice, Blast-1 in human, and OX-45 in the rat), a GPI-anchored member of the Ig superfamily. It is widely distributed on leukocytes, but not on non-hematopoietic cells, and its ligands include CD2 (LFA-2) and CD244 (2B4 antigen). The HM48-1 mAb blocks binding of soluble CD2 to CD48-bearing cells, blocks the interaction of CD2 and CD244, inhibits spleen cell proliferative responses to mitogens, augments the proliferative response of spleen cells when cross-linked with anti-CD3e mAbs, and inhibits priming of CTL in vitro. In vivo administration of HM48-1 antibody can prolong survival of cardiac allografts, an effect which is greatly enhanced by the addition of anti-CD2 mAb 12-15. This hamster mAb to a mouse leukocyte antigen does not cross-react with rat leukocytes.

The antibody was conjugated to BD Horizon™ Red 718, which has been developed exclusively by for BD Biosciences as a better alternative to Alexa Fluor™ 700. BD Horizon™ Red 718 can be excited by the red laser (628 – 640 nm) and, with an Em Max around 718 nm, it can be detected using a 730/45 nm filter. Due to similar excitation and emission properties, we do not recommend using R718 in combination with APC-R700 or Alexa Fluor™ 700.

567301 Rev. 1
Format Details
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R718
The BD Horizon™ Red 718 (R718) Dye is part of the BD red family of dyes. It is a small organic fluorochrome with an excitation maximum (Ex Max) at 695-nm and an emission maximum (Em Max) at 718-nm. Driven by BD innovation, R718 is designed to be excited by the red laser (627–640-nm) and detected using an optical filter centered near 720-nm (e.g., a 720/40-nm bandpass filter). R718 is a brighter alternative to Alexa Fluor™ 700. R718 is also a bright small molecule alternative to APC-R700 with lower spread into the APC detector. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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R718
Red 627-640 nm
695 nm
718 nm
567301 Rev.1
Citations & References
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Development References (5)

  1. Brown MH, Boles K, van der Merwe PA, Kumar V, Mathew PA, Barclay AN. 2B4, the natural killer and T cell immunoglobulin superfamily surface protein, is a ligand for CD48. J Exp Med. 1998; 188(11):2083-2090. (Biology). View Reference
  2. Kato K, Koyanagi M, Okada H, et al. CD48 is a counter-receptor for mouse CD2 and is involved in T cell activation. J Exp Med. 1992; 176(5):1241-1249. (Immunogen: Blocking, (Co)-stimulation, ELISA, Immunoprecipitation, Inhibition, Stimulation, Western blot). View Reference
  3. Latchman Y, McKay PF, Reiser H. Identification of the 2B4 molecule as a counter-receptor for CD48. J Immunol. 1998; 161(11):5809-5812. (Biology). View Reference
  4. Qin L, Chavin KD, Lin J, Yagita H, Bromberg JS. Anti-CD2 receptor and anti-CD2 ligand (CD48) antibodies synergize to prolong allograft survival. J Exp Med. 1994; 179(1):341-346. (Biology). View Reference
  5. Wong YW, Williams AF, Kingsmore SF, Seldin MF. Structure, expression, and genetic linkage of the mouse BCM1 (OX45 or Blast-1) antigen. Evidence for genetic duplication giving rise to the BCM1 region on mouse chromosome 1 and the CD2/LFA3 region on mouse chromosome 3. J Exp Med. 1990; 171(6):2115-2130. (Biology). View Reference
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567301 Rev. 1

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