BD, 554507, BD Pharmingen™ PE Rat Anti-Human GM-CSF

Alternative Name: CSF2; Colony stimulating factor 2 (granulocyte-macrophage); CSF; GMCSF
Reactivity: Human (QC Testing)
Isotype: Rat LEW, also known as Lewis IgG2a
Immunogen: Recombinant human GM-CSF
Application: Intracellular staining (flow cytometry) (Routinely Tested)
Concentration: 0.2 mg/ml
RRID: AB_395440
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 Immunofluorescent Staining and Flow Cytometric Analysis : The BVD2-21C11 antibody is useful for the immunofluorescent staining and flow cytometric analysis to identify and enumerate GM-CSF producing cells within mixed cell populations. (see Figure, Left panel ). 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, http://www.bdbiosciences.com/us/s/resources, and go to the protocols section under "Cytokines (Intracelullar Staining)". A useful control for demonstrating specificity of staining is either of the following: 1) pre-block the fluorochrome-conjugated BVD2-21C11 antibody with ligand (e.g., recombinant human GM-CSF, Cat No. 550068) prior to staining, or 2) pre-block the fixed/permeabilized cells with unlabeled BVD2- 21C11 antibody (Cat. No. 554503) prior to staining. The intracellular staining technique and use of blocking controls are described in detail by C. Prussin and D. Metcalfe.  A suitable rat IgG2a isotype control for assessing the level of background staining on paraformaldehyde-fixed/saponinpermeabilized mouse or human cells is PE-R35-95 immunoglobulin (Cat. No. 554689); use at comparable concentrations to antibody of interest (e..g., ≤0.5 µg mAb/1 million cells). ELISA Detection: The biotinylated BVD2-21C11 antibody (Cat. No. 554505) is useful as a detection antibody for a sandwich ELISA for measuring human GM-CSF protein levels.  For testing GM-CSF in serum or plasma, the OptEIA™ ELISA Set (Cat. No. 555126) is recommended. IP/WB: The purified BVD2-21C11 antibody has been reported to be useful for immunoprecipitation studies and for Western blotting.  Please note that this application is not routinely tested at BD Biosciences Pharmingen.
Product Notices: Product Notices Since applications vary, each investigator should titrate the reagent to obtain optimal results. An isotype control should be used at the same concentration as the antibody of interest. 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. Ficoll-Paque is a trademark of Amersham Biosciences Limited. Cy is a trademark of GE Healthcare. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.