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Multiparameter flow cytometric analysis of RP-1 Antigen expression on rat peripheral blood leucocytes. Rat whole blood was pretreated with BD Pharm Lyse™ Lysing Buffer (Cat No. 555899) to lyse erythrocytes. The leucocytes were washed and stained with APC Mouse IgG2a, κ Isotype Control (Cat. No. 550882; Left Plot) or APC Mouse Anti-Rat RP-1 Antigen (Cat. No. 566870; Right Plot) at 1 µg/test. The two-parameter pseudocolor density plot showing the correlated expression of RT-1 Antigen (or Ig Isotype control staining) versus side light-scatter signals was derived from gated events with the forward and side light-scatter characteristics of viable rat 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.
BD Pharmingen™ APC Mouse Anti-Rat RP-1 Antigen
Regulatory Status Legend
Any use of products other than the permitted use without the express written authorization of Becton, Dickinson and Company is strictly prohibited.
Preparation And Storage
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
- 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.
- This APC-conjugated reagent can be used in any flow cytometer equipped with a dye, HeNe, or red diode laser.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
Companion Products
The RP-1 monoclonal antibody specifically recognizes the RP-1 Antigen. This cell surface marker is expressed on rat peritoneal and peripheral blood neutrophils. Amongst bone marrow cells, the RP-1 Antigen is expressed on band form and mature neutrophils but is not expressed on promyelocytes, myelocytes, and metamyelocytes. The RP-1 antibody does not bind to either rat monocytes, macrophages, eosinophils or to peritoneal neutrophils from mice, rabbits, guinea pigs, or to human peripheral blood neutrophils. Expression of the RP-1 Antigen on rat peritoneal neutrophils is enhanced by cellular stimulation with Phorbol 12-Myristate 13-Acetate (PMA) or Concanavalin A (ConA). Immunoprecipitation and SDS-PAGE analysis of non-treated and PMA-activated rat neutrophil membranes with the RP-1 antibody revealed two main bands of approximately 85 kDa. The RP-1 antibody is also known as the Mouse Anti-Rat Granulocytes antibody.
Development References (3)
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Francis WR, Ireland RE, Spear AM, et al. Flow Cytometric Analysis of Hematopoietic Populations in Rat Bone Marrow. Impact of Trauma and Hemorrhagic Shock.. Cytometry A. 2019; 95(11):1167-1177. (Clone-specific: Flow cytometry, Fluorescence activated cell sorting). View Reference
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Gotoh S, Itoh M, Fujii Y, Arai S, Sendo F. Enhancement of the expression of a rat neutrophil-specific cell surface antigen by activation with phorbol myristate acetate and concanavalin A. J Immunol. 1986; 137(2):643-650. (Immunogen: Flow cytometry, Immunoprecipitation, Radioimmunoassay). View Reference
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Skrajnar S, Anzur Lasnik M, Bedina Zavec A. A flow cytometric method for determination of the blood neutrophil fraction in rats.. J Am Assoc Lab Anim Sci. 2009; 48(2):152-6. (Clone-specific: Flow cytometry). View Reference
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