AffiniPure F(ab’)2 Fragment Goat anti-Rabbit IgG(H+L), Alexa Fluor 647
Aqueous buffered solution containing protein stabilizer and ≤0.03% sodium azide.
To minimize cross-reactivity, these goat anti-rabbit IgG whole antibodies have been cross-adsorbed against bovine IgG, goat IgG, mouse IgG, rat IgG, and human IgG. Cross-adsorption or pre-adsorption is a purification step to increase specificity of the antibody resulting in higher sensitivity and less background staining. The secondary antibody solution is passed through a column matrix containing immobilized serum proteins from potentially cross-reactive species. Only the nonspecific-binding secondary antibodies are captured in the column, and the highly specific secondaries flow through. The benefits of this extra step are apparent in multiplexing/multicolor-staining experiments (e.g., flow cytometry) where there is potential cross-reactivity with other primary antibodies or in tissue/cell fluorescent staining experiments where there are may be the presence of endogenous immunoglobulins.
Flow cytometric analysis of anti-CD19 CAR expression on Jurkat cells. Jurkat cells were lentivirally transduced with anti-CD19 CAR and cultured. 5×105 cells were stained for the expression of anti-CD19 CAR with Rabbit Anti-Mouse FMC63 scFv Polyclonal Antibody (Cat. No. 500019, right panel). Secondary staining was carried out with AffiniPure F(ab’)2 Fragment Goat anti-Rabbit IgG(H+L), Alexa Fluor 647 (Cat. No. 700002). Non-transduced Jurkat cells were used as a control for gating of CAR expression (left panel).Acquisition of >10, 000 events was performed.
Preparation and Storage
Shipped at 2-8°C. Store undiluted at 2-8°C short term (4 weeks) and protected from prolonged exposure to light. Store at -20°C in small aliquots for long term storage. Avoid freeze/thaw cycle.
1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
2. 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.