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Alexa Fluor™ 647 Rat Anti-Mouse CD45
Alexa Fluor™ 647 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 either unstained [for a Fluorescence-Minus-One (FMO) control] or stained with Alexa Fluor™ 647 Rat Anti-Mouse CD45 antibody (Cat No. 567376; 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 FMO control) 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 either unstained [for a Fluorescence-Minus-One (FMO) control] or stained with Alexa Fluor™ 647 Rat Anti-Mouse CD45 antibody (Cat No. 567376; 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 FMO control) 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.

Product Details
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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 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. Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
  6. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  7. This product is provided under an intellectual property license between Life Technologies Corporation and BD Businesses. The purchase of this product conveys to the buyer the non-transferable right to use the purchased amount of the product and components of the product in research conducted by the buyer (whether the buyer is an academic or for-profit entity). The buyer cannot sell or otherwise transfer (a) this product (b) its components or (c) materials made using this product or its components to a third party or otherwise use this product or its components or materials made using this product or its components for Commercial Purposes. Commercial Purposes means any activity by a party for consideration and may include, but is not limited to: (1) use of the product or its components in manufacturing; (2) use of the product or its components to provide a service, information, or data; (3) use of the product or its components for therapeutic, diagnostic or prophylactic purposes; or (4) resale of the product or its components, whether or not such product or its components are resold for use in research. For information on purchasing a license to this product for any other use, contact Life Technologies Corporation, Cell Analysis Business Unit Business Development, 29851 Willow Creek Road, Eugene, OR 97402, USA, Tel: (541) 465-8300. Fax: (541) 335-0504.
  8. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  9. Alexa Fluor™ is a trademark of Life Technologies Corporation.
567376 Rev. 1
Antibody Details
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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).

567376 Rev. 1
Format Details
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Alexa Fluor™ 647
Alexa Fluor™ 647 Dye is part of the BD red family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 653-nm and an emission maximum (Em Max) at 669-nm. Alexa Fluor 647 is designed to be excited by the Red laser (627-640 nm) and detected using an optical filter centered near 520-nm (e.g., a 660/20 nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
Alexa Fluor™ 647
Red 627-640 nm
653 nm
669 nm
567376 Rev.1
Citations & References
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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
View All (5) View Less
567376 Rev. 1

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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.

Refer to manufacturer's instructions for use and related User Manuals and Technical Data Sheets before using this product 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.