FITC Rat Anti-Mouse IFN-γ
Clone XMG1.2 (RUO)
- Brand BD Pharmingen™
- Alternative Name Ifg; Ifng; IFN-γ; IFN-g; IFN-gamma; Interferon gamma; Type II Interferon
- Concentration 0.5 mg/ml
- Isotype Rat IgG1, κ
- Reactivity Mouse (QC Testing)
Intracellular staining (flow cytometry) (Routinely Tested)
- Immunogen Mouse IFN-γ Recombinant Protein
- Storage Buffer Aqueous buffered solution containing protein stabilizer and ≤0.09% sodium azide.
- Regulatory Status RUO
Regulatory Status Legend
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.
FITC, fluorescein isothiocyanate, is a fluorochrome with a molecular weight of 389 Da. FITC is sensitive to pH changes and photobleaching. Due to nearly identical excitation and emission properties but different spillover characteristics, FITC and Alexa Fluor® 488 cannot be used simultaneously. FITC is relatively dim and should be reserved for highly expressed markers whenever possible.
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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.
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- 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.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- An isotype control should be used at the same concentration as the antibody of interest.
Immunofluorescent Staining and Flow Cytometric Analysis: The FITC-conjugated XMG1.2 antibody can be used for multicolor immunofluorescent staining and flow cytometric analyses to identify and enumerate IFN-γ producing cells within mixed cell populations. For optimal immunofluorescent staining with flow cytometric analysis, this anti-cytokine antibody should be titrated (≤0.5 µg mAb/million cells).
A useful control for deomonstrating specificity of staining is either of the following: (1) pre-block the conjugated XMG1.2 antibody with a molar excess of ligand (e.g., recombinant mouse IFN-γ; Cat. No. 554587) prior to staining, or (2) pre-block the fixed/permeabilized cells with un-conjugated XMG1.2 antibody (Cat. No. 554409) prior to staining. The staining technique and use of blocking controls are described in detail by C. Prussin and D. Metcelfe. A suitable rat IgG1 isotype control for assessing the level of background staining on paraformaldehyde-fixed/saponin-permeabilized mouse cells is FITC-R3-34 (Cat. No. 554684); use at comparable concentrations to antibody of interest.