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PE Rat Anti-Mouse CD45
PE 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 PE Rat IgG2b, κ Isotype Control (Cat No. 553989, dashed line histogram) or PE Rat Anti-Mouse CD45 antibody (Cat No. 567111; 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 PE Rat IgG2b, κ Isotype Control (Cat No. 553989, dashed line histogram) or PE Rat Anti-Mouse CD45 antibody (Cat No. 567111; 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.

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
AB_2870024
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 unconjugated antibody and free dye were removed.

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 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. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  5. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  6. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
567111 Rev. 2
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).

        

567111 Rev. 2
Format Details
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PE
R-Phycoerythrin (PE), is part of the BD family of Phycobiliprotein dyes. This fluorochrome is a multimeric fluorescent phycobiliprotein with excitation maximum (Ex Max) of 496 nm and 566 nm and an emission maximum (Em Max) at 576 nm. PE is designed to be excited by the Blue (488 nm), Green (532 nm) and Yellow-Green (561 nm) lasers and detected using an optical filter centered near 575 nm (e.g., a 575/26-nm bandpass filter). As PE is excited by multiple lasers, this can result in cross-laser excitation and fluorescence spillover on instruments with various combinations of Blue, Green, and Yellow-Green lasers. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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PE
Yellow-Green 488 nm, 532 nm, 561 nm
496 nm, 566 nm
576 nm
567111 Rev.2
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
567111 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.

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.