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Biotin Mouse Anti-R-Phycoerythrin
Biotin Mouse Anti-R-Phycoerythrin
Amplification of PE-conjugated anti-mouse CD19 staining of BALB/c mouse splenocytes. Mouse splenocytes were incubated with PE Rat Anti-Mouse CD19 (Cat. No. 557399/553786) alone or followed by Biotin Mouse Anti-R-Phycoerythrin (Cat. No. 552603) and PE Streptavidin (Cat.  No. 554061, right panel). The overlayed histograms correspond to stained splenocytes (shaded histogram), unstained splenocytes (left panel, unshaded histogram), or splenocytes stained with Biotin Mouse Anti-R-Phycoerythrin and PE Streptavidin (right panel, unshaded histogram). The signal-to-noise ratio of median fluorescence intensity (MFI) of the CD19+ population to the CD19- population in both panels (shaded histograms) is 87 for Panel A and 172 for right panel, demonstrating an approximate two-fold specific amplification of staining. Data shown is for gated lymphocytes based on light scatter. Flow cytometry was performed on a BD FACSCalibur™ flow cytometry system.
Amplification of PE-conjugated anti-mouse CD19 staining of BALB/c mouse splenocytes. Mouse splenocytes were incubated with PE Rat Anti-Mouse CD19 (Cat. No. 557399/553786) alone or followed by Biotin Mouse Anti-R-Phycoerythrin (Cat. No. 552603) and PE Streptavidin (Cat.  No. 554061, right panel). The overlayed histograms correspond to stained splenocytes (shaded histogram), unstained splenocytes (left panel, unshaded histogram), or splenocytes stained with Biotin Mouse Anti-R-Phycoerythrin and PE Streptavidin (right panel, unshaded histogram). The signal-to-noise ratio of median fluorescence intensity (MFI) of the CD19+ population to the CD19- population in both panels (shaded histograms) is 87 for Panel A and 172 for right panel, demonstrating an approximate two-fold specific amplification of staining. Data shown is for gated lymphocytes based on light scatter. Flow cytometry was performed on a BD FACSCalibur™ flow cytometry system.
Product Details
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BD Pharmingen™
Mouse IgG1, κ
R-phycoerythrin
Flow cytometry (Routinely Tested)
0.5 mg/ml
AB_394440
Aqueous buffered solution containing ≤0.09% sodium azide.
RUO


Preparation And Storage

The antibody was conjugated with biotin under optimum conditions, and unreacted biotin was removed. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.

Recommended Assay Procedures

For the amplification of PE fluorescence of PE-conjugated antibodies, it is recommended that biotinylated E31-1459 be used as a secondary antibody followed by PE-conjugated Streptavidin (Cat. No. 554061).

Product Notices

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  2. 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.
  3. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  4. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
552603 Rev. 3
Antibody Details
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E31-1459

The E31-1459 antibody reacts with R-Phycoerythrin (PE), a commonly used fluorochrome for flow cytometry, alone or as a tandem conjugate with other fluorochromes (eg, PE-Cy™7). The binding of the E31-1459 antibody to PE does not quench the fluorescence of the PE molecule. The use of biotinylated E31-1459 in a three-step staining procedure with Streptavidin-PE and a PE-conjugated primary antibody can specifically amplify the PE fluorescence.

552603 Rev. 3
Format Details
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Biotin
Biotin is a ubiquitous co-factor (also known as Vitamin B7) that has many properties that make it extremely useful for molecular biology. Biotin has an extremely high affinity for the Avidin family of proteins (Kd = 10-15 M), making it the perfect tool to link two molecules. Biotin labeled antibodies can be combined with any number of Avidin-conjugated probes in order to customize an assay to a particular need. This is especially useful in the case of magnetic cell separation using streptavidin/magnetic bead conjugates, or in the case of flow cytometry using streptavidin/fluorophore conjugates.
Biotin
552603 Rev.3

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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.