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Alexa Fluor® 647 Mouse IgG1 κ Isotype control
Alexa Fluor® 647 Mouse IgG1 κ Isotype control
Flow cytometric analysis on U-937 cells. U-937 cells (ATCC CRL-1593) were serum starved overnight (RPMI containing 0.1% FCS). The following day cells were either left untreated (solid line) or treated (dotted line) with recombinant human IFN (Cat. No. 554617; 1000 U/ml for 10 minutes at 37°C). Cells were then fixed in 2% paraformaldehyde (10 minutes at 37°C) and permeabilized by adding 90% methanol, (BD Phosflow™ Perm Buffer III, Cat. No. 558050) to the cell pellet (30 minutes on ice or overnight at at -20°C). Cells were washed twice in BD Pharmingen™ Stain Buffer (Cat. No. 554656) and stained with Alexa Fluor® 647 Isotype Control antibody (1 hour at RT; Cat. No. 557783). The cells were analyzed on a BD FACSCalibur™ flow cytometry system.
Flow cytometric analysis on U-937 cells. U-937 cells (ATCC CRL-1593) were serum starved overnight (RPMI containing 0.1% FCS). The following day cells were either left untreated (solid line) or treated (dotted line) with recombinant human IFN (Cat. No. 554617; 1000 U/ml for 10 minutes at 37°C). Cells were then fixed in 2% paraformaldehyde (10 minutes at 37°C) and permeabilized by adding 90% methanol, (BD Phosflow™ Perm Buffer III, Cat. No. 558050) to the cell pellet (30 minutes on ice or overnight at at -20°C). Cells were washed twice in BD Pharmingen™ Stain Buffer (Cat. No. 554656) and stained with Alexa Fluor® 647 Isotype Control antibody (1 hour at RT; Cat. No. 557783). The cells were analyzed on a BD FACSCalibur™ flow cytometry system.
Product Details
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BD Phosflow™
IgG1, kappa; Ighg1, Igh-4, γg1; Igk-C, IGKC, Ig κ
Mouse IgG1, κ
Intracellular staining (flow cytometry), Isotype control (Routinely Tested)
20 µl
Aqueous buffered solution containing BSA and ≤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 to the dye under optimum conditions and unreacted dye was removed.

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

  1. This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test).
  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. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  5. Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
  6. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  7. This product is provided under an intellectual property license between Life Technologies Corporation and BD Businesses. The purchase of this product conveys to the buyer the non-transferable right to use the purchased amount of the product and components of the product in research conducted by the buyer (whether the buyer is an academic or for-profit entity). The buyer cannot sell or otherwise transfer (a) this product (b) its components or (c) materials made using this product or its components to a third party or otherwise use this product or its components or materials made using this product or its components for Commercial Purposes. Commercial Purposes means any activity by a party for consideration and may include, but is not limited to: (1) use of the product or its components in manufacturing; (2) use of the product or its components to provide a service, information, or data; (3) use of the product or its components for therapeutic, diagnostic or prophylactic purposes; or (4) resale of the product or its components, whether or not such product or its components are resold for use in research. For information on purchasing a license to this product for any other use, contact Life Technologies Corporation, Cell Analysis Business Unit Business Development, 29851 Willow Creek Road, Eugene, OR 97402, USA, Tel: (541) 465-8300. Fax: (541) 335-0504.
  8. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  9. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  10. Alexa Fluor™ is a trademark of Life Technologies Corporation.
557783 Rev. 7
Antibody Details
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MOPC-21

The MOPC-21 immunoglobulin is a mouse myeloma protein. The MOPC-21 immunoglobulin was selected as an isotype control following screening for low background on a variety of mouse and human tissues.

557783 Rev. 7
Format Details
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Alexa Fluor™ 647
Alexa Fluor™ 647 Dye is part of the BD red family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 653-nm and an emission maximum (Em Max) at 669-nm. Alexa Fluor 647 is designed to be excited by the Red laser (627-640 nm) and detected using an optical filter centered near 520-nm (e.g., a 660/20 nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
Alexa Fluor™ 647
Red 627-640 nm
653 nm
669 nm
557783 Rev.7
Citations & References
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Development References (1)

  1. Prussin C, Metcalfe DD. Detection of intracytoplasmic cytokine using flow cytometry and directly conjugated anti-cytokine antibodies. J Immunol Methods. 1995; 188(1):117-128. (Clone-specific: Blocking). View Reference
557783 Rev. 7

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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.