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BV421 Rat Anti-Mouse CD18
BV421 Rat Anti-Mouse CD18
Flow cytometric analysis using BD OptiBuild™ BV421 Rat Anti-Mouse CD18 antibody (Cat. No. 744597; solid line histogram) on C57BL/6 mouse splenocytes, with Isotype Control (dotted line histogram). Flow cytometry was performed using a BD LSRFortessa™ X-20  Flow Cytometer System.
Flow cytometric analysis using BD OptiBuild™ BV421 Rat Anti-Mouse CD18 antibody (Cat. No. 744597; solid line histogram) on C57BL/6 mouse splenocytes, with Isotype Control (dotted line histogram). Flow cytometry was performed using a BD LSRFortessa™ X-20  Flow Cytometer System.
Product Details
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BD OptiBuild™
Cd18; ITGB2; Integrin β2 chain
Mouse (Tested in Development)
Rat IgG2a, κ
Mouse CTL glycoproteins
Flow cytometry (Qualified)
0.2 mg/ml
AB_2742346
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 that minimize unconjugated dye and antibody.

Recommended Assay Procedures

BD™ CompBeads can be used as surrogates to assess fluorescence spillover (Compensation).  When fluorochrome conjugated antibodies are bound to 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 CompBead to ensure that BD Comp beads are appropriate for your specific cellular application.

For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime two or more BD Horizon Brilliant dyes are used in the same experiment.  Fluorescent dye interactions may cause staining artifacts which may affect data interpretation.  The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions.  More information can be found in the Technical Data Sheet of the BD Horizon Brilliant Stain Buffer (Cat. No. 563794/566349) or the BD Horizon Brilliant Stain Buffer Plus (Cat. No. 566385).

Product Notices

  1. The production process underwent stringent testing and validation to assure that it generates a high-quality conjugate with consistent performance and specific binding activity. However, verification testing has not been performed on all conjugate lots.
  2. Researchers should determine the optimal concentration of this reagent for their individual applications.
  3. An isotype control should be used at the same concentration as the antibody of interest.
  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. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  6. BD Horizon Brilliant Stain Buffer is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,575,303; 8,354,239.
  7. BD Horizon Brilliant Violet 421 is covered by one or more of the following US patents: 8,158,444; 8,362,193; 8,575,303; 8,354,239.
  8. Pacific Blue™ is a trademark of Molecular Probes, Inc., Eugene, OR.
  9. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  10. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
744597 Rev. 3
Antibody Details
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M18/2

The M18/2 antibody specifically recognizes the common β2 chain of LFA-1 (CD11a/CD18, αLβ2 integrin), Mac-1 (CD11b/CD18, αMβ2 integrin), and gp150, 95 (CD11c/CD18, αXβ2 integrin). Expression of CD18 is limited to leukocytes, where it is widely distributed in consort with the three integrin α chains (CD11a, CD11b, and CD11c). Among splenocytes, NK cells have the highest density of CD18, and T lymphocytes express a higher density than the remaining cells. The β2 integrins are important mediators of leukocyte-endothelium interactions. It has been reported that M18/2 antibody blocks in vivo metastasis of the LB lymphoma to the spleen and that it blocks in vitro formation of aggregates of LB cells and splenocytes. However, other reports indicate that mAb M18/2 has no effect on CTL-mediated killing, adherence of C3bi-sensitized erythrocytes to Mac-1, antigen-specific binding of T cells to antigen-producing cells, or rejection of cardiac allografts. Recent in vitro studies indicate that M18/2 antibody stimulates adhesion of Mac-1 to its ligands C3bi and ICAM-1, and it stimulates adhesion of LFA-1 to ICAM-1, but it has no effect upon the interactions of LFA-1 with ICAM-2 nor ICAM-3.

The antibody was conjugated to BD Horizon BV421 which is part of the BD Horizon Brilliant™ Violet family of dyes. With an Ex Max near 407 nm and Em Max near 421 nm, BD Horizon BV421 can be excited by the violet laser (405 nm) and detected with a 450/50 nm filter. BD Horizon BV421 conjugates are very bright, often exhibiting a 10 fold improvement in brightness compared to Pacific BlueTM conjugates. Due to nearly identical excitation and emission properties but different spillover characteristics, BD Horizon BV421, Pacific Blue, and BD Horizon V450 cannot be used simultaneously.

744597 Rev. 3
Format Details
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BV421
The BD Horizon Brilliant Violet™ 421 (BV421) Dye is part of the BD Horizon Brilliant Violet™ family of dyes. This polymer-technology based dye has an excitation maximum (Ex Max) of 407-nm and an emission maximum (Em Max) at 423-nm. Driven by BD innovation, BV421 is designed to be excited by the violet laser (405-nm) and detected using an optical filter centered near 420-nm (e.g., a 431/28-nm or 450/50-nm bandpass filter). BV421 is an ideal alternative for V450 as it is approximately ten times brighter with less spillover into the BV510/V500 detector. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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BV421
Violet 405 nm
407 nm
423 nm
744597 Rev.3
Citations & References
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Development References (8)

  1. Driessens MH, van Hulten P, Zuurbier A, La Riviere G, Roos E. Inhibition and stimulation of LFA-1 and Mac-1 functions by antibodies against murine CD18. Evidence that the LFA-1 binding sites for ICAM-1, -2, and -3 are distinct. J Leukoc Biol. 1996; 60(6):758-765. (Clone-specific: Blocking, Stimulation). View Reference
  2. Ishida Y, Chused TM, Murakami S, Abe R. Antigen-specific cell conjugate formation and long-lasting calcium responses in recognition of Mls cellular superantigen by cloned murine T lymphocytes. Cell Immunol. 1994; 155(2):414-427. (Clone-specific: Blocking, Stimulation). View Reference
  3. 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
  4. Sanchez-Madrid F, Simon P, Thompson S, Springer TA. Mapping of antigenic and functional epitopes on the alpha- and beta-subunits of two related mouse glycoproteins involved in cell interactions, LFA-1 and Mac-1. J Exp Med. 1983; 158(2):586-602. (Immunogen: Immunoprecipitation, Western blot). View Reference
  5. Springer TA. Adhesion receptors of the immune system. Nature. 1990; 346(6283):425-434. (Biology). View Reference
  6. Springer TA. Traffic signals for lymphocyte recirculation and leukocyte emigration: the multistep paradigm. Cell. 1994; 76(2):301-314. (Biology). View Reference
  7. Zahalka MA, Naor D. Beta 2-integrin dependent aggregate formation between LB T cell lymphoma and spleen cells: assessment of correlation with spleen invasiveness. Int Immunol. 1994; 6(6):917-924. (Clone-specific: Blocking, Stimulation). View Reference
  8. Zahalka MA, Okon E, Naor D. Blocking lymphoma invasiveness with a monoclonal antibody directed against the beta-chain of the leukocyte adhesion molecule (CD18). J Immunol. 1993; 150(10):4466-4477. (Clone-specific: Blocking, Stimulation). View Reference
View All (8) View Less
744597 Rev. 3

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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.