FITC Rat Anti-Human IL-6

BD Pharmingen™ FITC Rat Anti-Human IL-6

Name: FITC Rat Anti-Human IL-6
Brand: BD Pharmingen™
SKU: 554696

クローン MQ2-6A3

(RUO)

Expression of IL-6 by stimulated CD14+ human monocytes. Human PBMC were stimulated for 6 hours with LPS (100 ng/ml final concentration) in the presence of BD GolgiStop™ (2 µM final concentration; Cat. No. 554724). The PBMC were harvested, stained with PE-mouse anti-human CD14 monoclonal antibody (PE-M5E2, Cat. No. 555398), fixed, permeabilized, and subsequently stained with 0.25 µg of FITC- rat anti-human IL-6 antibody (FITC-MQ2- 6A3, Cat. No. 554696), following BD Pharmingen's staining protocol (left panel). The data reflect gating on monocytes, based on forward and side scattered light signals. To demonstrate specificity of staining, the binding by the FITC-MQ2-6A3 antibody was blocked by preincubation of the conjugated antibody with recombinant human IL-6 protein (0.5 µg, Cat. No. 550071; middle panel) and by preincubation of the fixed/permeabilized cells with unlabeled MQ2-6A3 antibody (5.0 µg; Cat. No. 559068, right panel) prior to staining with the FITC-MQ2-6A3 antibody. The quadrant markers for the bivariate dot plots were set based on the autofluorescence controls and verified using the recombinant cytokine blocking and unlabeled antibody blocking specificity control.

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製品詳細

Brand: BD Pharmingen™

Reactivity: Human (QC Testing)

Isotype: Rat IgG2a, κ

Immunogen: Recombinant human IL-6

Application: Intracellular staining (flow cytometry) (Routinely Tested)

Concentration: 0.5 mg/ml

RRID: AB_395514

Storage Buffer: Aqueous buffered solution containing protein stabilizer and ≤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 with FITC under optimum conditions, and unreacted FITC was removed.

推奨アッセイ手順

Flow cytometry: The MQ2-6A3 antibody is useful for immunofluorescent staining and flow cytometric analysis to identify and enumerate human IL-6 producing cells within mixed cell populations. FITC-conjugated MQ2-6A3 antibody (Cat. No. 554696) is especially suitable for these experiments (see figure). For optimal immunofluorescent staining for flow cytometric analysis, this anti-cytokine antibody should be titrated (≤ 0.5 µg mAb/million cells). A useful control for demonstrating specificity of staining is either of the following: 1) preblock the FITC- MQ2-6A3 antibody with 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. A suitable rat IgG2a isotype control for assessing the level of background staining on paraformaldehyde-fixed/saponin-permeabilized human cells is FITC-R35-95 (Cat. No. 554688).

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.
Use of these products to measure activation antigens expressed on mononuclear cell subsets for the purpose of monitoring immunoregulatory status can fall under one or more claims of the following patents: US Patent Nos. 5,445,939, 5,656,446, 5,843,689; European Patent No. 319,543; Canadian Patent No. 1,296,622; Australian Patent No. 615,880; and Japanese Patent No. 2,769,156.
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.
Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.

554696 Rev. 2

抗体の詳細

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.

554696 Rev. 2

フォーマットの詳細

FITC

Fluorescein (FITC) is part of the BD blue family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 494-nm and an emission maximum (Em Max) at 518-nm. FITC is designed to be excited by the Blue laser (488-nm) and detected using an optical filter centered near 520 nm (e.g., a 530/30-nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.

フォーマット: FITC

励起源: Blue 488 nm

最大励起波長: 494 nm

最大蛍光波長: 518 nm

554696 Rev.2

引用＆参考文献

Development References (5)

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. (Biology). View Reference
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.
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.
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
Sander B, Andersson J, Andersson U. Assessment of cytokines by immunofluorescence and the paraformaldehyde-saponin procedure. Immunol Rev. 1991; 119:65-93. (Biology). View Reference

すべて表示する (5)

554696 Rev. 2

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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.