Flow cytometric analysis of CD45 expression on rat splenocytes. Lewis rat splenic leucocytes were preincubated with Purified Mouse Anti-Rat CD32 antibody (Rat BD Fc Block™) (Cat. No. 550270/550271). The cells were then stained with either Alexa Fluor® 647 Mouse IgG1 κ Isotype Control (Cat. No. 557714; dashed line histogram) or Alexa Fluor® 647 Mouse Anti-Rat CD45 (Cat. No. 565465; solid line histogram). 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 splenocytes. Flow cytometric analysis was performed using a BD LSRFortessa™ Cell Analyzer System.
Flow cytometric analysis of CD45 expression on rat splenocytes. Lewis rat splenic leucocytes were preincubated with Purified Mouse Anti-Rat CD32 antibody (Rat BD Fc Block™) (Cat. No. 550270/550271). The cells were then stained with either Alexa Fluor® 647 Mouse IgG1 κ Isotype Control (Cat. No. 557714; dashed line histogram) or Alexa Fluor® 647 Mouse Anti-Rat CD45 (Cat. No. 565465; solid line histogram). 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 splenocytes. Flow cytometric analysis was performed using a BD LSRFortessa™ Cell Analyzer System.
Product Details
BD Pharmingen™
Ptprc; Lca; Leucocyte common antigen; RT7; T200
Rat (QC Testing)
Mouse BALB/c IgG1, κ
Leukocyte Common Antigen-enriched Glycoprotein Fraction from Wistar Rat Thymocytes
Flow cytometry (Routinely Tested)
0.2 mg/ml
24699
AB_2739250
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 Alexa Fluor® 647 under optimum conditions, and unreacted Alexa Fluor® 647 was removed.
Product Notices
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- An isotype control should be used at the same concentration as the antibody of interest.
- 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.
- Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
- The Alexa Fluor®, Pacific Blue™, and Cascade Blue® dye antibody conjugates in this product are sold under license from Molecular Probes, Inc. for research use only, excluding use in combination with microarrays, or as analyte specific reagents. The Alexa Fluor® dyes (except for Alexa Fluor® 430), Pacific Blue™ dye, and Cascade Blue® dye are covered by pending and issued patents.
- Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
Companion Products
Stain Buffer (FBS) RUO
Size
500 mL
Cat No.
554656
Stain Buffer (BSA) RUO
Size
500 mL
Cat No.
554657
Lysing Buffer RUO
Size
100 mL
Cat No.
555899
Alexa Fluor® 647 Mouse IgG1 κ Isotype Control RUO
Size
100 Tests
Cat No.
557714
Purified Mouse Anti-Rat CD32 RUO
Size
0.1 mg
Cat No.
550270
Purified Mouse Anti-Rat CD32 RUO
Size
0.5 mg
Cat No.
550271
565465 Rev. 1
Antibody Details
OX-1
The OX-1 antibody recognizes all molecular forms of CD45 (Leukocyte Common Antigen) on all hematopoietic cells except erythrocytes. CD45 is a member of the Protein Tyrosine Phosphatase (PTP) family; its intracellular (COOH-terminal) region contains two PTP catalytic domains while the extracellular region is highly variable due to alternative splicing of exons 4, 5, and 6 (designated A, B, and C, respectively), and differing levels of glycosylation. The CD45 isoforms detected in rat are cell type-, maturation-, and activation state-specific. The CD45 isoforms play complex roles in T-cell and B-cell antigen receptor signal transduction.
565465 Rev. 1
Format Details
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
565465 Rev.1
Citations & References
Development References (3)
-
Johnson P, Maiti A, Ng DHW. CD45: A family of leukocyte-specific cell surface glycoproteins. In: Herzenberg LA, Weir DM, Herzenberg LA, Blackwell C , ed. Weir's Handbook of Experimental Immunology, Vol 2. Cambridge: Blackwell Science; 1997:62.1-62.16.
-
Sunderland CA, McMaster WR, Williams AF. Purification with monoclonal antibody of a predominant leukocyte-common antigen and glycoprotein from rat thymocytes. Eur J Immunol. 1979; 9(2):155-159. (Immunogen: Flow cytometry). View Reference
-
Woollett GR, Barclay AN, et al. Molecular and antigenic heterogeneity of the rat leukocyte common antigen from thymocytes and T and B lymphocytes. . Eur J Immunol. 1985; 15:168-173. (Biology).
565465 Rev. 1
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.
