BD, 562586, BD Phosflow™ PE Mouse anti-Smad2 (pS465/pS467)/Smad3 (pS423/pS425)

Alternative Name: SMAD2, SMAD3; MADH2, MADH3; MAD homolog 2, MAD homolog 3
Reactivity: Human (QC Testing), Mouse (Tested in Development), Rat (Predicted)
Isotype: Mouse IgG1, κ
Immunogen: Phosphorylated Human Smad2 Peptide
Application: Intracellular staining (flow cytometry) (Routinely Tested)
Vol. Per Test: 5 µl
RRID: AB_11151915
Storage Buffer: Aqueous buffered solution containing BSA and ≤0.09% sodium azide.
Regulatory Status: RUO
Preparation And Storage: 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 R-PE under optimum conditions, and unconjugated antibody and free PE were removed.
Recommended Assay Procedures: Recommended Assay Procedures This antibody conjugate is suitable for intracellular staining of human lymphoid cell lines, peripheral blood mononuclear cells, and mouse splenocytes using BD Cytofix™ Fixation Buffer or BD Phosflow™ Lyse/Fix Buffer with BD Phosflow™ Perm Buffer III (see table, below). Prior to stimulation, cells were serum starved overnight at a density of 2-10 million cells/mL in flat-bottom 96- or 6-well tissue culture plates or in lo osely capped, round-bottom tubes containing approximately 100 μ L of cells in suspension. Note: 1. Serum starvation for 2 hours following PBMC isolation was not sufficient to reduce basal phosphorylation of Smad2 and Smad3. 2. Do not mix or agitate untreated cells until just before the cells are ready to be fixed, since agitation of serum-starved mouse or human primary leukocytes prior to fixation increased Smad2/3 phosphorylation, even in the absence of exogenous TGF-β.
Product Notices: Product Notices 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). For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors. 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. Source of all serum proteins is from USDA inspected abattoirs located in the United States.