Flow cytometric analysis of CD48 on mouse splenocytes. Splenocytes from BALB/c mice were stained either with APC-Cy7 Hamster IgG1, λ1 Isotype Control (Cat. No.561206; dashed line histogram) or with the APC-Cy7 Hamster Anti-Mouse CD48 antibody (Cat. No. 561242; solid line histogram). Flow cytometric histograms were derived from gated events with the forward and side light-scattering characteristics of viable cells. Flow cytometry was performed on a BD™ LSR II Flow Cytometry System.
Flow cytometric analysis of CD48 on mouse splenocytes. Splenocytes from BALB/c mice were stained either with APC-Cy7 Hamster IgG1, λ1 Isotype Control (Cat. No.561206; dashed line histogram) or with the APC-Cy7 Hamster Anti-Mouse CD48 antibody (Cat. No. 561242; solid line histogram). Flow cytometric histograms were derived from gated events with the forward and side light-scattering characteristics of viable cells. Flow cytometry was performed on a BD™ LSR II Flow Cytometry System.
Product Details
BD Pharmingen™
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
AB_10644381
Aqueous buffered solution containing protein stabilizer 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 with APC-Cy7 under optimum conditions, and unconjugated antibody and free APC-Cy7 were removed.
Product Notices
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- APC-Cy7 is a tandem fluorochrome composed of Allophycocyanin (APC), which is excited by laser lines between 595 and 647 nm and serves as an energy donor, coupled to the cyanine dye Cy7™, which acts as an energy acceptor and fluoresces at 780 nm. BD Biosciences Pharmingen has maximized the fluorochrome energy transfer in APC-Cy7, thus maximizing its fluorescence emission intensity, minimizing residual emission from APC, and minimizing required electronic compensation in multilaser-laser flow cytometry systems. Note: Although every effort is made to minimize the lot-to-lot variation in residual emission from APC, it is strongly recommended that every lot be tested for differences in the amount of compensation required and that individual compensation controls are run for each APC-Cy7 conjugate.
- APC-Cy7 tandem fluorochrome emission is collected in a detector for fluorescence wavelengths of 750 nm and higher.
- Please observe the following precautions: Absorption of visible light can significantly alter the energy transfer occurring in any tandem fluorochrome conjugate; therefore, we recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to prevent exposure of conjugated reagents, including cells stained with those reagents, to room illumination.
- Cy is a trademark of Amersham Biosciences Limited.
- Warning: Some APC-Cy7 and PE-Cy7 conjugates show changes in their emission spectrum with prolonged exposure to formaldehyde. If you are unable to analyze fixed samples within four hours, we recommend that you use BD™ Stabilizing Fixative (Cat. No. 338036).
- 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.
- 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.
- An isotype control should be used at the same concentration as the antibody of interest.
561242 Rev. 2
Antibody Details
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.
561242 Rev. 2
Format Details
APC-Cy7
APC-Cy7 dye is a part of the BD APC red family of dyes. This tandem fluorochrome is comprised of a Allophycocyanin (APC) donor that has excitation maxima (Ex Max) of 651 nm and an acceptor dye, Cy™7, with an emission maximum (Em Max) at 779 nm. APC-Cy7 can be excited by the Red (627-640 nm) laser and detected using an optical filter centered near 780 nm (e.g., a 760/60 nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
APC-Cy7
Red 627-640 nm
651 nm
779 nm
561242 Rev.2
Citations & References
Development References (5)
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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
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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). View Reference
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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
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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
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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
561242 Rev. 2
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.
