Brand: BD Pharmingen
CSF2; Colony stimulating factor 2 (granulocytes-macrophages); CSF; GMCSF
Reactivity: Humans (quality control tests)
LEW rat, also known as Lewis IgG2a
Recombinant human GM-CSF
Intracellular staining (flow cytometry) (tested routinely)
Concentration: 0.2mg / ml
Buffered aqueous solution containing ≤ 0.09% sodium azide.
Regulatory status: RUO
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 to R-PE under optimal conditions and unconjugated antibody and free PE were removed.
Recommended test procedures
Immunofluorescent Staining and Flow Cytometry Analysis: BVD2-21C11 antibody is useful for 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 cytometry analysis, this anti-cytokine antibody should be titrated (≤0.5 µg mAb / million cells). For the specific methodology, visit our website, http://www.bdbiosciences.com/us/s/resources, and go to the protocols section under “Cytokines (intracellular staining)”.
A useful control to demonstrate specificity of staining is any of the following: 1) pre-block fluorochrome-conjugated antibody BVD2-21C11 with ligand (eg, recombinant human GM-CSF, catalog number 550068) prior to staining, or 2) pre-blocking cells fixed / permeabilized with unlabeled BVD2-21C11 antibody (cat. # 554503) prior to staining.
The intracellular staining technique and the 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 in paraformaldehyde fixed / saponin permeabilized mouse or human cells is immunoglobulin PE-R35-95 (Cat. # 554689); use at concentrations comparable to the antibody of interest (eg, ≤ 0.5 µg mAb / 1 million cells).
ELISA Detection: Biotinylated BVD2-21C11 Antibody (Cat. # 554505) is useful as a detection antibody for a sandwich ELISA to measure human GM-CSF protein levels. For testing GM-CSF in serum or plasma, the OptEIA ™ ELISA Kit (Cat. # 555126) is recommended.
IP / WB: 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 by BD Biosciences Pharmingen.
- Since applications vary, each investigator should titrate the reagent for optimal results.
- An isotype control should be used at the same concentration as the antibody of interest.
- Caution: Sodium azide produces highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in tap water before disposal to avoid accumulation of potentially explosive deposits in pipes.
- Ficoll-Paque is a trademark of Amersham Biosciences Limited.
- Cy is a trademark of GE Healthcare.