BB700 Mouse Anti-Rat RP-1 Antigen

BD Horizon™ BB700 Mouse Anti-Rat RP-1 Antigen

Name: BB700 Mouse Anti-Rat RP-1 Antigen
Brand: BD Horizon™
SKU: 566871

クローン RP-1

(RUO)

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™ BB700 Mouse IgG2a, κ Isotype Control (Cat. No. 566419; Left Plot) or BD Horizon BB700 Mouse Anti-Rat RP-1 Antigen antibody (Cat. No. 566871; 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.

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製品詳細

Brand: BD Horizon™

Alternative Name: RP1 Antigen; Rat Granulocytes; Rat Neutrophils

Reactivity: Rat (QC Testing)

Isotype: Mouse BALB/c IgG2a, κ

Immunogen: WKA/Hok rat peritoneal neutrophils

Application: Flow cytometry (Routinely Tested)

Concentration: 0.2 mg/ml

RRID: AB_2869924

Storage Buffer: Aqueous buffered solution containing ≤0.09% sodium azide.

Regulatory Status: 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.

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 two washes after staining with antibodies. Cells may be prepared, stained with antibodies and washed twice with wash buffer per established protocols for immunofluorescent 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

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.
For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
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.
BD Horizon Brilliant Blue 700 is covered by one or more of the following US patents: 8,455,613 and 8,575,303.
Cy is a trademark of GE Healthcare.
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.

関連製品

Stain Buffer (FBS) RUO

サイズ 500 mL カタログ番号 554656

Stain Buffer (BSA) RUO

サイズ 500 mL カタログ番号 554657

Brilliant Stain Buffer RUO

サイズ 100 Tests カタログ番号 563794

Brilliant Stain Buffer RUO

サイズ 1000 Tests カタログ番号 566349

Brilliant Stain Buffer Plus RUO

サイズ 1000 Tests カタログ番号 566385

Lysing Buffer RUO

サイズ 100 mL カタログ番号 555899

詳細を表示

566871 Rev. 1

抗体の詳細

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 BB700, which is part of the BD Horizon Brilliant™ Blue family of dyes. It is a polymer-based tandem dye developed exclusively by BD Biosciences. With an excitation max of 485 nm and an emission max of 693 nm, BD Horizon BB700 can be excited by the 488 nm laser and detected in a standard PerCP-Cy™5.5 set (eg, 695/40-nm filter). This dye provides a much brighter alternative to PerCP-Cy5.5 with less cross laser excitation off the 405 nm and 355 nm lasers.

566871 Rev. 1

フォーマットの詳細

BB700

The BD Horizon Brilliant™ Blue 700 (BB700) Dye is part of the BD Horizon Brilliant™ Blue family of dyes. This tandem fluorochrome is comprised of a polymer-technology dye donor with an excitation maximum (Ex Max) of 476-nm and an acceptor dye with an emission maximum (Em Max) at 695-nm. Driven by BD innovation, BB700 is designed to be excited by the blue laser (488-nm) and detected using an optical filter centered near 695-nm (e.g., a 695/20-nm bandpass filter). The donor dye can be excited by the Violet (405 nm) laser and the acceptor dye can be excited by the red (627–640 nm) laser resulting in cross-laser excitation and fluorescence spillover. BB700 Reagents are significantly brighter than equivalent PerCP or PerCP-Cy5.5 reagents and are less sensitive to photobleaching. In addition, BB700 shows much less excitation by the violet (407-nm) laser resulting in less spillover. BB700 has minimal yellow green (562-nm) excitation and is ideal for instruments with both blue (488-nm) and yellow green (562-nm) lasers. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.

フォーマット: BB700

励起源: Blue 488 nm

最大励起波長: 476 nm

最大蛍光波長: 695 nm

566871 Rev.1

引用＆参考文献

Development References (3)

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
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
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

566871 Rev. 1

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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.