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PerCP-Cy5.5 Rat Anti-Mouse CD45
PerCP-Cy5.5 Rat Anti-Mouse CD45
Flow cytometric analysis of CD45 expression on mouse splenocytes. Splenic leucocytes from BALB/c mice were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142). The cells were stained with either PerCP-Cy5.5 Rat IgG2b, κ Isotype Control (Cat No. 550764; dashed line histogram) or PerCP-Cy5.5 Rat Anti-Mouse CD45 antibody (Cat No. 567310; 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. The fluorescence histogram showing CD45 expression (or Ig Isotype control staining) was derived from gated events with the forward and side light-scatter characteristics of viable (DAPI-negative) mouse leucocytes. 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.
Flow cytometric analysis of CD45 expression on mouse splenocytes. Splenic leucocytes from BALB/c mice were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142). The cells were stained with either PerCP-Cy5.5 Rat IgG2b, κ Isotype Control (Cat No. 550764; dashed line histogram) or PerCP-Cy5.5 Rat Anti-Mouse CD45 antibody (Cat No. 567310; 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. The fluorescence histogram showing CD45 expression (or Ig Isotype control staining) was derived from gated events with the forward and side light-scatter characteristics of viable (DAPI-negative) mouse leucocytes. 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.
製品詳細
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BD Pharmingen™
Cd45; L-CA; LCA; Ly-5; Ly5; Ptprc
Mouse (QC Testing)
Rat LEW, also known as Lewis IgG2b, λ
Mouse Lymphoma Cell Line
Flow cytometry (Routinely Tested)
0.2 mg/ml
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 with PerCP-Cy5.5 under optimum conditions, and unconjugated antibody and free PerCP-Cy5.5 were removed. Storage of PerCP-Cy5.5 conjugates in unoptimized diluent is not recommended and may result in loss of signal intensity.

推奨アッセイ手順

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 cell and CompBead to ensure that BD Comp beads 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. 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.
  6. PerCP-Cy5.5–labelled antibodies can be used with FITC- and R-PE–labelled reagents in single-laser flow cytometers with no significant spectral overlap of PerCP-Cy5.5, FITC, and R-PE fluorescence.
  7. PerCP-Cy5.5 is optimized for use with a single argon ion laser emitting 488-nm light. Because of the broad absorption spectrum of the tandem fluorochrome, extra care must be taken when using dual-laser cytometers, which may directly excite both PerCP and Cy5.5™. We recommend the use of cross-beam compensation during data acquisition or software compensation during data analysis.
  8. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  9. Cy is a trademark of Global Life Sciences Solutions Germany GmbH or an affiliate doing business as Cytiva.
  10. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).

関連製品

Purified Rat Anti-Mouse CD16/CD32 (Mouse BD Fc Block™) RUO
サイズ 0.5 mg カタログ番号 553142
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Purified Rat Anti-Mouse CD16/CD32 (Mouse BD Fc Block™) RUO
サイズ 0.1 mg カタログ番号 553141
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Stain Buffer (BSA) RUO
サイズ 500 mL カタログ番号 554657
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Stain Buffer (FBS) RUO
サイズ 500 mL カタログ番号 554656
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Lysing Buffer RUO
サイズ 100 mL カタログ番号 555899
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PerCP-Cy™5.5 Rat IgG2b, κ Isotype Control RUO
サイズ 0.1 mg カタログ番号 550764
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567310 Rev. 1
抗体の詳細
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I3/2.3

The mAb I3/2.3 recognizes mouse CD45, which is also known as the leukocyte common antigen (L-CA or LCA), T200, or Lymphocyte antigen 5 (Ly-5). CD45 is a 180-240 kDa single-pass type I transmembrane glycoprotein that is encoded by Ptprc (protein tyrosine phosphatase, receptor type, C) which belongs to the protein tyrosine phosphatase (PTP) family. CD45 is variably expressed on thymocytes and other hematopoietic cells except mature erythrocytes and platelets. Different isoforms of CD45 arise from alternative splicing of variable exons 4, 5, and 6, which encode A, B, and C determinants, respectively, that encode extracellular domains of the transmembrane receptor. The differential expression of these isoforms is related to the cell's lineage as well as its state of activation and maturation. The mAb I3/2.3 reportedly recognizes all CD45 isoforms. CD45 plays key roles as a protein tyrosine-protein phosphatase in leucocyte responses including signal pathways transduced by antigen specific receptors expressed on T cells (TCRs) and B cells (BCRs).

567310 Rev. 1
フォーマットの詳細
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PerCP-Cy5.5
PerCP-Cy5.5 dye is part of the BD blue family of dyes. This tandem fluorochrome is comprised of a fluorescent protein complex (PerCP) with an excitation maximum (Ex Max) of 482 nm and an acceptor dye with an emission maximum (Em Max) at 676 nm. PerCP-Cy5 is designed to be excited by the blue laser (488-nm) and detected using an optical filter centered near 680 nm (e.g., a 695/40 nm bandpass filter). The donor dye can be partially excited by the Violet (405-nm) laser resulting in cross-laser excitation and fluorescence spillover. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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PerCP-Cy5.5
Blue 488 nm
482 nm
676 nm
567310 Rev.1
引用&参考文献
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Development References (5)

  1. Alaverdi N. Monoclonal antibodies to mouse cell-surface antigens.. Curr Protoc Immunol. 2002; Appendix 4:Appendix 4B. (Clone-specific). View Reference
  2. Johnson P, Greenbaum L, Bottomly K, Trowbridge IS. Identification of the alternatively spliced exons of murine CD45 (T200) required for reactivity with B220 and other T200-restricted antibodies. J Exp Med. 1989; 169(3):1179-1184. (Clone-specific: Flow cytometry). View Reference
  3. Lefrancois L, Goodman T. Developmental sequence of T200 antigen modifications in murine T cells.. J Immunol. 1987; 139(11):3718-24. (Clone-specific: Flow cytometry, Immunoprecipitation, Radioimmunoassay). View Reference
  4. Takedachi M, Qu D, Ebisuno Y, et al. CD73-generated adenosine restricts lymphocyte migration into draining lymph nodes.. J Immunol. 2008; 180(9):6288-96. (Biology). View Reference
  5. Trowbridge IS. Interspecies spleen-myeloma hybrid producing monoclonal antibodies against mouse lymphocyte surface glycoprotein, T200.. J Exp Med. 1978; 148(1):313-23. (Immunogen: Immunoprecipitation, Ribonuclease protection assay). View Reference
すべて表示する (5) 表示項目を減らす
567310 Rev. 1

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