Skip to main content Skip to navigation
Alexa Fluor® 488 Rat Anti-Mouse IFN-γ
Alexa Fluor® 488 Rat Anti-Mouse IFN-γ

Expression of IFN-γ  by stimulated CD4+ and CD4-BALB/c spleen cells. Splenocytes from BALB/C mice were stimulated for 4 hrs with PMA (5 ng/ml, Sigma Cat No P-8139) and Ionomycin (500 ng, Sigma Cat No I-0634) in the presence of Brefeldin A (GolgiPlug, Cat. No. 555029). Cells were harvested, fixed, permeabilized and stained with PE rat anti-mouse CD4 (PE-RM4-5, Cat. No. 553048) and either rat anti-mouse IFN-γ (Alexa Fluor® 488-XMG1.2, Cat. No. 557724), (left panel) or immunoglobulin isotype control (Alexa Fluor® 488-R3-34, Cat. No. 557720), (right panel) by using the BD Pharmingen staining protocol. To demonstrate specificity of staining the binding of Alexa Fluor® 488-XMG1.2 was blocked by preincubation of the fixed/permeabilized cells with an excess of unlabelled XMG1.2 antibody (5 µg, Cat. No. 554409, data not shown) prior to stainining. The quadarant markers for the bivariate dot plots were set based on the autofluorescence and isotype controls.

Expression of IFN-γ  by stimulated CD4+ and CD4-BALB/c spleen cells. Splenocytes from BALB/C mice were stimulated for 4 hrs with PMA (5 ng/ml, Sigma Cat No P-8139) and Ionomycin (500 ng, Sigma Cat No I-0634) in the presence of Brefeldin A (GolgiPlug, Cat. No. 555029). Cells were harvested, fixed, permeabilized and stained with PE rat anti-mouse CD4 (PE-RM4-5, Cat. No. 553048) and either rat anti-mouse IFN-γ (Alexa Fluor® 488-XMG1.2, Cat. No. 557724), (left panel) or immunoglobulin isotype control (Alexa Fluor® 488-R3-34, Cat. No. 557720), (right panel) by using the BD Pharmingen staining protocol. To demonstrate specificity of staining the binding of Alexa Fluor® 488-XMG1.2 was blocked by preincubation of the fixed/permeabilized cells with an excess of unlabelled XMG1.2 antibody (5 µg, Cat. No. 554409, data not shown) prior to stainining. The quadarant markers for the bivariate dot plots were set based on the autofluorescence and isotype controls.

製品詳細
Down Arrow Up Arrow


BD Pharmingen™
Mouse (QC Testing)
Rat IgG1, κ
Mouse IFN-γ Recombinant Protein
Intracellular staining (flow cytometry) (Routinely Tested)
0.2 mg/ml
AB_396832
Aqueous buffered solution containing ≤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 to Alexa Fluor® 488 under optimum conditions, and unreacted Alexa Fluor® 488 was removed. Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.

推奨アッセイ手順

Recommended Assay Procedure:

Immunofluorescent Staining and Flow Cytometric Analysis: The XMG1.2 antibody is useful for the immunofluorescent staining and flow cytometric analysis to identify and enumerate IFN-γ producing cells within mixed cell populations. The Alexa Fluor® 488-conjugated XMG1.2 antibodies (Cat. No. 557724) is especially suitable for these studies. For optimal immunofluorescent staining with flow cytometric analysis, this anti-cytokine antibody should be titrated.  A useful control for demonstrating specificity of staining is to pre-block the fixed/permabilized cells with unlabeled XMG1.2 antibody (Cat. No. 554409) prior to staining.  A suitable rat IgG1 isotype control for assessing the level of background staining on paraformaldehyde-fixed/saponin-permeabilized mouse or human cells is R3-34 immunoglobulin (Cat. No. 557720). Use at comparable concentrations to antibody of interest.

Product Notices

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  2. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  3. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  4. Alexa Fluor® 488 fluorochrome emission is collected at the same instrument settings as for fluorescein isothiocyanate (FITC).
  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. The Alexa Fluor®, Pacific Blue™, and Cascade Blue® dye antibody conjugates in this product are sold under license from Molecular Probes, Inc. for research use only, excluding use in combination with microarrays, or as analyte specific reagents. The Alexa Fluor® dyes (except for Alexa Fluor® 430), Pacific Blue™ dye, and Cascade Blue® dye are covered by pending and issued patents.
  7. Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
557724 Rev. 2
抗体の詳細
Down Arrow Up Arrow
XMG1.2

The XMG1.2 monoclonal antibody specifically binds to mouse interferon-γ (IFN-γ) protein. IFN-γ is a pleiotropic cytokine, of approximately 15-17 kDa, involved in the regulation of inflammatory and immune responses. It plays an important role in activation, growth, and differentiation of T and B lymphocytes, macrophages, NK cells and other non-hematopoietic cell types. IFN-γ production is associated with the Th1 cell differentiation. The purified form of this antibody has been reported to be a neutralizing antibody.

557724 Rev. 2
フォーマットの詳細
Down Arrow Up Arrow
Alexa Fluor™ 488
Alexa Fluor™ 488 Dye 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 517-nm. Alexa Fluor™ 488 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.
Alexa Fluor™ 488
Blue 488 nm
494 nm
517 nm
557724 Rev.2
引用&参考文献
Down Arrow Up Arrow

Development References (4)

  1. Abrams JS, Roncarolo MG, Yssel H, Andersson U, Gleich GJ, Silver JE. Strategies of anti-cytokine monoclonal antibody development: immunoassay of IL-10 and IL-5 in clinical samples. Immunol Rev. 1992; 127:5-24. (Clone-specific). View Reference
  2. Cherwinski HM, Schumacher JH, Brown KD, Mosmann TR. Two types of mouse helper T cell clone. III. Further differences in lymphokine synthesis between Th1 and Th2 clones revealed by RNA hybridization, functionally monospecific bioassays, and monoclonal antibodies. J Exp Med. 1987; 166(5):1229-1244. (Clone-specific). View Reference
  3. 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: Flow cytometry). View Reference
  4. Sander B, Hoiden I, Andersson U, Moller E, Abrams JS. Similar frequencies and kinetics of cytokine producing cells in murine peripheral blood and spleen. Cytokine detection by immunoassay and intracellular immunostaining. J Immunol Methods. 1993; 166(2):201-214. (Clone-specific). View Reference
すべて表示する (4) 表示項目を減らす
557724 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.