BD Pharmingen™ Alexa Fluor® 700 Rat anti-Mouse TNF 558000

Reactivity: Mouse (QC Testing)
Isotype: Rat IgG1
Immunogen: Recombinant Mouse TNF
Application: Intracellular staining (flow cytometry) (Routinely Tested)
Concentration: 0.2 mg/ml
RRID: AB_396980
Storage Buffer: Aqueous buffered solution containing protein stabilizer and ≤0.09% sodium azide.
Regulatory Status: RUO
Preparation And Storage: Preparation And Storage The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated to Alexa Fluor® 700 under optimum conditions, and unreacted Alexa Fluor® 700 was removed. Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.
Recommended Assay Procedures: Recommended Assay Procedures Immunofluorescent Staining and Flow Cytometry: The Alexa  Fluor® 700-conjugated MP6-XT22 antibody can be used for multicolor immunofluorescent staining and flow cytometric analyses to identify and enumerate TNF producing cells within mixed cell populations (see figure). For optimal immunofluorescent staining with flow cytometric analysis, this anti-cytokine antibody should be titrated (≤ 0.5 µg mAb/million cells). For specific methodology, please visit our web site, www.bdbiosciences.com, and go to the protocols section or the chapter on intracellular staining in the Immune Function Handbook. A useful control for demonstrating specificity of staining is either of the following: 1) pre-block the conjugated MP6-XT22 antibody with ligand (e.g., recombinant mouse TNF; Cat. No 554589) prior to staining, or 2) pre-block the fixed/permeabilized cells with unlabeled MP6-XT22 antibody (Cat. No 554416) prior to staining. The staining technique and use of blocking controls are described in detail by C. Prussin and D. Metcalfe. A suitable rat IgG1 isotype control for assessing the level of background staining on paraformaldehyde-fixed/saponin-permeabilized mouse cells is Alexa Fluor® 700 Rat IgG1 κ Isotype Control (Cat. No 558001); use at comparable concentrations to antibody of interest.
Product Notices: Product Notices 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. Alexa Fluor® 700 has an adsorption maximum of ~700nm and a peak fluorescence emission of ~720nm. Before staining cells with this reagent, please confirm that your flow cytometer is capable of exciting the fluorochrome and discriminating the resulting fluorescence. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors. 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. 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. 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. Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.