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F(ab')2 fragment antibodies are generated by pepsin digestion of whole IgG antibodies to remove most of the Fc region while leaving some of the hinge region. F(ab')2 fragments have two antigen-binding Fab portions linked together by disulfide bonds and therefore they are divalent. The average molecular weight is about 110 kDa. They are used for specific applications, such as to avoid binding of secondary antibodies to live cells with Fc receptors or to Protein A or Protein G.
Storage and Rehydration: Store freeze-dried solid at 2-8°C. Rehydrate with the indicated volume of dH2O and centrifuge if not clear. Store at 2-8°C – do not freeze. Prepare working dilution on day of use.
Expiration date: six months from date of rehydration. The expiration date may be extended if test results are acceptable for the intended use.
PerCP is a fluorescent peridinin-chlorophyll-protein complex isolated from dinoflagellates. We offer the form found in Dinophyceae sp. with a molecular weight of about 35.5 kDa. It has a broad spectrum of excitation with a main peak at 482 nm, and a long Stokes shift to an emission peak at 677 nm.
PerCP, Alexa Fluor® 488 (or FITC), and R-PE are excited at 488 nm with an argon laser, and thus can be used for one-, two-, and three-color analyses with single-laser flow cytometers. APC and Alexa Fluor® 647 are excited at 633 nm to give a fourth color with dual-laser flow cytometers.
PerCP is also highly water soluble, has a relatively low isoelectric point, and lacks potentially sticky carbohydrates.
It should be noted that the relatively high molecular weight of PerCP may preclude its use in procedures requiring good penetration into cells and tissues. It is predominantly intended for surface labeling of cells for flow cytometry.
2um thick FFPE sections of normal human lymph nodes were heated 2h @60C, followed by autoclaving for 15 in Trilogy Solution (CellMarque, USA) and 5 min incubation in a second vial of Trilogy Solution @98C. Slides were washed for 5 mins in 1x PBS/0.1% TritonX100 and blocked in 1x PBS/0.1% TritonX100/2% BSA for 15 min @37C. Primary antibodies against CD3 (1:200, Thermo Scientific, RM-9107-s1, rabbit ) and CD20 (1:50, eBioscience, 50-0202, eFluor660-conjugated) were diluted in 1x PBS/0.1% TritonX100/1% BSA and incubated overnight @4C. After 3x10min washes the slides were incubated in PerCP-conjugated anti-rabbit secondary antibody (1:100, Jackson Immunoresearch, 711-126-152) for 1h @RT and again washed 3x10 mins. DNA was counterstained with DAPI (Invitrogen) for 5 mins and mounted in PrologGold (Invitrogen, P36931) or Supermount (BioVision, 1211-20) antifade media.
Karina Silina - University of Zurich
This product is for in vitro research use only. It is not a medical device and it is not intended for diagnostic or therapeutic purposes.