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PE Rat Anti-Human IL-6
製品詳細
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BD Pharmingen™
Human (QC Testing)
Rat IgG2a, κ
Recombinant human IL-6
Intracellular staining (flow cytometry) (Routinely Tested)
20 µl
Aqueous buffered solution containing BSA and ≤0.09% sodium azide.
RUO


Preparation and Storage

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

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Immunofluorescent Staining and Flow Cytometric Analysis: The MQ2-6A3 antibody is useful for immunoflourescent staining and flow cytometric analysis to identify and enumerate human IL-6 producing cells within mixed cell populations. This 100 Test Size formulation of the PE-conjugated MQ2-6A3 antibody has been pre-titrated to assure effective intracellular detection of human IL-6 using 20 µl/1 x 10e6 cells.

A useful control for demonstrating specificity of staining is either of the following: 1) pre-block the conjugated MQ2-6A3 antibody with a ligand (e.g., recombinant human IL-6; Cat. No. 550071) prior to staining, or 2) pre-block the fixed/permeabilized cells with unlabelled MQ2-6A3 antibody (Cat. No. 559068) prior to staining. The staining technique and blocking controls are described in detail by C. Prussin and D. Metcalfe. A suitable rat IgG2a isotype control for assessing the level of background staining on paraformaldehyde-fixed/saponin-permeabilized human cells is also available in a 100 Test Size formulation PE-R35-95 (Cat. No. 559317).

Important Note: This pre-titered antibody solution does not contain a cell permeabilization agent. It is necessary to include a cell permeabilization agent when using the pre-titered antibody solution to stain fixed and permeabilized cells. Perm/Wash™ Buffer (Cat. No. 554723) contains the permeabilization agent saponin and is useful for this purpose as described in the USAGE section below.

USAGE

1. Resuspend 1 x 10e6 fixed and permeabilized cells in 20 µl of the pre-titered antibody solution and 30 µl of 1X Perm/Wash™ Buffer (Cat. No. 554723).

2. Incubate the cell suspension for 15 minutes (at RT or 4°C).

3. Wash twice in 100 µl of 1X Perm/Wash™ Buffer (Cat. No. 554723).

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. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  3. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  4. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  5. 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.
  6. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
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抗体の詳細
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MQ2-6A3

The MQ2-6A3 antibody reacts with human interleukin-6 (IL-6). The immunogen used to generate the MQ2-6A3 hybridoma was recombinant human IL-6. This is a neutralizing antibody.

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フォーマットの詳細
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PE
R-Phycoerythrin (PE), is part of the BD family of Phycobiliprotein dyes. This fluorochrome is a multimeric fluorescent phycobiliprotein with excitation maximum (Ex Max) of 496 nm and 566 nm and an emission maximum (Em Max) at 576 nm. PE is designed to be excited by the Blue (488 nm), Green (532 nm) and Yellow-Green (561 nm) lasers and detected using an optical filter centered near 575 nm (e.g., a 575/26-nm bandpass filter). As PE is excited by multiple lasers, this can result in cross-laser excitation and fluorescence spillover on instruments with various combinations of Blue, Green, and Yellow-Green lasers. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
altImg
PE
Yellow-Green 488 nm, 532 nm, 561 nm
496 nm, 566 nm
576 nm
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引用&参考文献
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Development References (4)

  1. Andersson J, Abrams J, Bjork L, et al. Concomitant in vivo production of 19 different cytokines in human tonsils. Immunology. 1994; 83(1):16-24. (Clone-specific). View Reference
  2. Andersson U, Andersson J. Immunolabeling of cytokine-producing cells in tissues and in suspension. In: Fradelizie D, Emelie D, ed. Cytokine Producing Cells. Paris: Inserm; 1994:32-49.
  3. Litton M, Andersson J, Bjork L, Fehniger T, Ulfgren AK, Andersson U. Cytoplasmic cytokine staining in individual cells. In: Debets and Savelkoul, ed. Human Cytokine Protocols. Humana Press; 1996.
  4. 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. (Methodology: IC/FCM Block). View Reference
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559331 Rev. 1

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.