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BB515 Goat Anti-Rat Ig
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This SKU will be discontinuing Apr 2024. Suggested alternate SKU is [554016] or for additional support, contact your local applications specialist. Contact Us #

BD Horizon™ BB515 Goat Anti-Rat Ig

Clone Polyclonal

(RUO)
BB515 Goat Anti-Rat Ig
Flow cytometric analysis of CD8b.2 or CD45R/B220 expression on mouse splenocytes. Mouse splenic leucocytes were either not stained with a primary antibody (control staining; dashed line histograms) or stained with Purified Rat Anti-Mouse CD8b.2 antibody (Cat. No. 553038; Left Panel; solid line histogram) or Purified Rat Anti-Mouse CD45R/B220 antibody (Cat. No. 557390; Right Panel: solid line histogram). The cells were washed and secondarily stained with BD Horizon BB515 Goat Anti-Rat Ig (Cat. No. 565012). The fluorescence histograms showing either CD8b.2 or CD45R/B220 expression (or no primary antibody control staining) were derived from gated events with the forward and side light-scatter characteristics of viable leucocytes. Flow cytometric analysis was performed using a BD LSRFortessa™ Cell Analyzer System.
Flow cytometric analysis of CD8b.2 or CD45R/B220 expression on mouse splenocytes. Mouse splenic leucocytes were either not stained with a primary antibody (control staining; dashed line histograms) or stained with Purified Rat Anti-Mouse CD8b.2 antibody (Cat. No. 553038; Left Panel; solid line histogram) or Purified Rat Anti-Mouse CD45R/B220 antibody (Cat. No. 557390; Right Panel: solid line histogram). The cells were washed and secondarily stained with BD Horizon BB515 Goat Anti-Rat Ig (Cat. No. 565012). The fluorescence histograms showing either CD8b.2 or CD45R/B220 expression (or no primary antibody control staining) were derived from gated events with the forward and side light-scatter characteristics of viable leucocytes. Flow cytometric analysis was performed using a BD LSRFortessa™ Cell Analyzer System.
Product Details
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BD Horizon™
Rat (QC Testing)
Goat Ig
Flow cytometry (Routinely Tested)
0.2 mg/ml
AB_2739041
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.

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 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. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  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 www.bdbiosciences.com/us/s/resources for technical protocols.
565012 Rev. 2
Antibody Details
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Poly1271

  BD Horizon™ BB515 Goat Anti-Rat Ig is intended to be a second-step reagent for immunofluorescent staining of cells pre-stained with Rat Ig primary antibodies. It also stains rat B cells and has little reactivity with rat non-B cells or mouse splenocytes. As a second step, it is reactive with rat IgG and IgM monoclonal antibodies although a weaker signal may be detected when the primary antibody has a rat IgM or IgG2b isotype due to its adsorption with mouse Ig. It has weak crossreactivity detectable by flow cytometry with some, but not all, hamster immunoglobulins.

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.

565012 Rev. 2
Format Details
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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.
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BB515
Blue 488 nm
490 nm
515 nm
565012 Rev.2
Citations & References
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565012 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.