BD, 554462, BD Pharmingen™ PE Rat Anti-Mouse CD126

Alternative Name: Il6ra; IL-6 Receptor α chain; IL-6RA; IL-6R alpha
Reactivity: Mouse (QC Testing)
Isotype: Rat IgG2b, κ
Immunogen: OKT4 hybridoma cells
Application: Flow cytometry (Routinely Tested)
Concentration: 0.2 mg/ml
Workshop Number: none
RRID: AB_395408
Storage Buffer: Aqueous buffered solution containing ≤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 The PE conjugated D7715A7 antibody can be used for the immunofluorescent staining and flow cytometric analyses of mouse peripheral blood leukocytes and cell lines which express IL-6 receptors. 1. To help block nonspecific staining due to Fc receptors, preincubate ~1 million cells with 1 µg of the purified anti-CD32/CD16 (anti-FcγII/III receptor) antibody (clone 2.4G2, Fc Block™, Cat. No. 553142/553143) for 30 minutes at 4°C. 2.  Incubate the cells with 0.12 - 2.0 µg of PE conjugated D7715A7 antibody (Cat. No. 554462) at 4°C for 45 minutes. Wash cells three times with staining medium containing sodium azide ( e.g ., Dulbecco's PBS or tissue culture medium [without phenol red] with 0.09% sodium azide and 1% heat-inactivated FCS).  We encourage investigators to titrate the D7715A7 antibody up to saturating levels for optimal performance, minimizing the risk for dim staining. 3.  Resuspend cells in staining medium and analyze by flow cytometry using appropriate specificity and compensation controls. Using this method, positive staining was seen with the B9 cell line but not with the mouse MC/9 mast cell line and C20.4 CD4+ T cell line.
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. 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.