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BB515 Mouse Anti-Rat RP-1 Antigen
BB515 Mouse Anti-Rat RP-1 Antigen
Multiparameter flow cytometric analysis of RP-1 Antigen expression on rat peripheral blood leucocytes. Rat whole blood was treated with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899) to lyse erythrocytes. The leucocytes were washed and stained with either BD Horizon™ BB515 Mouse IgG2a, κ Isotype Control (Cat. No. 564515; Left Plot) or BD Horizon BB515 Mouse Anti-Rat RP-1 Antigen antibody (Cat. No. 566872; Right Plot) at 1 µg/test. A two-parameter pseudocolor density plot showing the correlated expression of RP-1 Antigen (or Ig Isotype control staining) versus side-light scatter (SSC-A) signals was derived from gated events with the forward and side-light scatter characteristics of viable leucocyte populations. 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.
Multiparameter flow cytometric analysis of RP-1 Antigen expression on rat peripheral blood leucocytes. Rat whole blood was treated with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899) to lyse erythrocytes. The leucocytes were washed and stained with either BD Horizon™ BB515 Mouse IgG2a, κ Isotype Control (Cat. No. 564515; Left Plot) or BD Horizon BB515 Mouse Anti-Rat RP-1 Antigen antibody (Cat. No. 566872; Right Plot) at 1 µg/test. A two-parameter pseudocolor density plot showing the correlated expression of RP-1 Antigen (or Ig Isotype control staining) versus side-light scatter (SSC-A) signals was derived from gated events with the forward and side-light scatter characteristics of viable leucocyte populations. 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 Horizon™
RP1 Antigen; Rat Granulocytes; Rat Neutrophils
Rat (QC Testing)
Mouse BALB/c IgG2a, κ
WKA/Hok rat peritoneal neutrophils
Flow cytometry (Routinely Tested)
0.2 mg/ml
AB_2869925
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 BD Horizon™ BB515 under optimum conditions and unconjugated antibody was removed.

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 cells and CompBead to ensure that BD Comp beads are appropriate for your specific cellular application.

For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime two or more BD Horizon Brilliant dyes are used in the same experiment.  Fluorescent dye interactions may cause staining artifacts which may affect data interpretation.  The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions.  More information can be found in the Technical Data Sheet of the BD Horizon Brilliant Stain Buffer (Cat. No. 563794/566349) or the BD Horizon Brilliant Stain Buffer Plus (Cat. No. 566385).

For optimal results, it is recommended to perform 2 washes after staining with antibodies. Cells may be prepared, stained with antibodies and washed twice with wash buffer per established protocols for immunofluorescence staining, prior to acquisition on a flow cytometer. Performing fewer than the recommended wash steps may lead to increased spread of the negative population.

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. The manufacture, use, sale, offer for sale, or import of this product is subject to one or more patents or pending applications. This product, and only in the amount purchased by buyer, may be used solely for buyer’s own internal research, in a manner consistent with the accompanying product literature. No other right to use, sell or otherwise transfer (a) this product, or (b) its components is hereby granted expressly, by implication or by estoppel. Diagnostic uses require a separate license.
  6. BD Horizon Brilliant Stain Buffer is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,575,303; 8,354,239.
  7. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  8. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
566872 Rev. 1
抗体の詳細
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RP-1

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.

The antibody was conjugated to BD Horizon BB515 which is part of the BD Horizon Brilliant™ Blue family of dyes. With an Ex Max near 490 nm and an Em Max near 515 nm, BD Horizon BB515 can be excited by the blue laser (488 nm) laser and detected with a 530/30 nm filter. This dye has been exclusively developed by BD Biosciences and is up to seven times brighter than FITC with less spillover into the PE channel. Due to similar excitation and emission properties, BB515, FITC, and Alexa Fluor® 488 cannot be used simultaneously. It is not recommended to use BB515 in cocktails that include Streptavidin conjugates as it may cause high background.

566872 Rev. 1
フォーマットの詳細
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BB515
The BD Horizon Brilliant™ Blue 515 (BB515) dye is part of the BD Horizon Brilliant™ Blue family of dyes. This dye is a polymer fluorochrome with an excitation maximum (Ex Max) at 490-nm and an emission maximum (Em Max) of 515-nm. Driven by BD innovation, BB515 is designed to be excited by the blue laser (488-nm) and detected using an optical filter centered near 520-nm (e.g., 530/30-nm). BB515 reagents are significantly brighter than equivalent FITC or Alexa Fluor™ 488 reagents with less spillover into the PE detector. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
altImg
BB515
Blue 488 nm
490 nm
515 nm
566872 Rev.1
引用&参考文献
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Development References (3)

  1. 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
  2. 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
  3. 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
566872 Rev. 1

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