In indirect staining, the primary antibody is not fluorochrome-labeled, but is detected by a fluorochrome-labeled secondary antibody. This second reagent may be an antibody with specificity for the first antibody. Alternatively, the avidin-biotin system can be used, whereby an antibody is conjugated to biotin and detected with fluorochrome-labeled avidin.
With the wide range of conjugated antibodies now available, this method means that unconjugated primary antibodies raised against many different targets can be used in conjunction with a labeled secondary antibody for FACS analysis. This widens the choice of target proteins for the researcher.
When using secondary antibodies - All the primary antibodies have been raised in different species to ensure cross labeling via the secondary antibodies is not encountered. The secondary antibodies recognize one of the species exclusively.
• High sensitivity: More than one labeled antibody is bound per antigen molecule; The signal intensity could be amplified on a single epitope.
• Flexible: Different primary antibodies can be used with a single labeled secondary antibody; The reservoir of primary antibodies suitable for flow cytometry would be broadened for no limitation of fluorochrome labeling.
• Cost-saving: Fewer labeled antibodies are required.
|Human 14-3-3 beta / YWHAB Antibody||Human B7-DC / PD-L2 / CD273 Antibody|
|Human 14-3-3 sigma / Stratifin / YWHAS Antibody||Human B7-H3 / CD276 Antibody|
|Mouse 4-1BBL / CD137L / TNFSF9 Antibody||Human BACE1 Antibody|
|Human 4E-BP1 / EIF4EBP1 Antibody||Human Bcl-XL / BCL2L1 Antibody|
|AcMNPV Envelope glycoprotein gp64 / AcmNPV-gp64 Antibody||Human BID Antibody|
|Human ACY1 / Aminoacylase-1 Antibody||Human BLyS / TNFSF13B / BAFF Antibody|
|Human ADAM15 / MDC15 Antibody||Human BMP-4 Antibody|
|Human AK4 / Adenylate Kinase 4 / AK3L1 Antibody||Human c-MET / HGFR Antibody|
|Human AKR1B1 Antibody||Human C1QBP / HABP1 Antibody|
|Human ALDH1 / ALDH1A1 / ALDC Antibody||Human C4.4A / LYPD3 Antibody|
|Human ALDH7A1 Antibody||Human CALML3 Antibody|
|Human Angiotensinogen / SerpinA8 / AGT Antibody||Human Carbonic Anhydrase XII / CA12 Antibody|
|Influenza Anti-H7N9 Hemagglutinin / HA Antibody||Human Caspase-14 / CASP14 Antibody|
|Human Apolipoprotein H / APOH Antibody||Human CD111 / Nectin-1 / PVRL1 Antibody|
|Human ARD1 Antibody||Human CD116 / GM-CSFR Antibody|
|Human ASAM / CLMP Antibody||Human CD131 / CSF2RB / IL3RB / IL5RB Antibody|
|Human ASF1B Antibody||Mouse CD132 / IL2RG Antibody|
|Human ASGR2 / ASGPR2 Antibody||Human CD14 Antibody|
|Human B2M / beta-2 microglobulin Antibody|