Multi-epitope coverage with the precision of monoclonals

Invitrogen™ Superclonal™ secondary antibodies represent a recombinant antibody technology designed to provide precise and accurate detection of mouse, rabbit and goat primary antibodies in a variety of applications. The proprietary screening and production process yields specific mixtures of recombinant goat or rabbit secondary antibodies that bind with the epitope-precision of monoclonal antibodies, while also achieving the multi-epitope coverage (e.g., H+L) and sensitivity of polyclonal antibodies. Superclonal secondary antibodies are also animal origin–free after the first immunization. Each Superclonal secondary antibody is formulated and optimized to help achieve excellent results in ELISA, western blot, and cell imaging.

“Superclonal results were better than expected. Specifically, the Superclonal did not exhibit any background or noise whatsoever. Really bright and vibrant resolution on the staining. I was confident that the signals were specific to the targets and not the antibody background.”

—Nathan Schurman, Research Diabetes Technician


Superclonal secondary antibodies help enable precise and accurate detection

Invitrogen™ Superclonal™ secondary antibodies are designed to accurate and precise results. The endogenous tubulin of U-87 MG (human neuroblastoma) cells were labeled with anti-alpha tubulin primary antibody, which was then detected with Goat anti-Mouse IgG (H+L) Superclonal Secondary Antibody, Alexa Fluor 488 conjugate antibody (green). Simultaneously, DISC1 was labeled with Goat anti-Rabbit IgG (H+L) Superclonal Secondary Antibody, Alexa Fluor 555 conjugate (red). Cell nuclei (blue) were stained with DAPI (blue).


Features of Superclonal secondary antibodies

  • Developed as recombinant monoclonal antibodies to enable precise and accurate detection
  • Formulated to recognize both heavy- and light-chain epitopes (H+L) of target IgG molecules
  • Selected and optimized for use with cell imaging, ELISA, and western blotting applications
  • Offered in four types: goat anti-mouse (GAM), goat anti-rabbit (GAR), rabbit anti-mouse (RAM), rabbit anti-goat (RAG)
  • Available unconjugated and conjugated with biotin, horseradish peroxidase (HRP), and selected Alexa Fluor™ dyes
 
 

Lot-to-lot consistency

Representative images demonstrating lot-to-lot consistency of performance of Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor™ 488 conjugate in immunocytochemistry. No primary control to assess background is shown. Endogenous HIF-1 alpha in U20S human osteocarcinoma cells was labeled (green) with Rabbit anti-HIF-1 alpha primary antibody, and visualized using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor™ 488 conjugate (0.4 µg/mL, 1:2500). Nuclei (blue) were stained with SlowFade™ Gold Antifade Mountant using DAPI (1:50). F-actin (red) was stained with Alexa Fluor™ 594 Phalloidin (1:200). 


Compare the technology

Superclonal secondary antibodies compared to traditional secondary antibodies

  Polyclonal antibodies
(pAb)
Superclonal™ secondary antibodies Monoclonal antibodies (mAb)
How they are made Affinity purify (by positive and/or negative selection) antibodies from the serum of immunized animals Produce and characterize numerous individual AOF recombinant antibodies, then carefully screen, select, and pool specific ones Isolate and select a desired antibody-producing cell from host immunized animal, then culture it to produce a single antibody clone
What they are A large, undefined pool of antibodies from the host serum, selected by affinity purification but otherwise having unknowable sets of specific epitope and binding-affinity characteristics Precisely characterized sets of specific AOF recombinant antibodies having known, complementary sets of epitopes and affinity binding features Well characterized single antibody clones having one specific epitope and affinity binding features.
Epitope coverage and signal amplification

Excellent

Broad epitope coverage for the target IgG (e.g., heavy and light chain, H+L) ensures good sensitivity and signal amplification for the target primary antibody

Excellent

Selected and pooled to provide complementary, specific and optimal epitope coverage (e.g., H+L) and binding quality to maximize performance in key applications

Poor

Binds to a single epitope Can select for a clone that has no off-target binding (i.e., background or cross-reactivity)

Specificity and lot-to-lot consistency

Poor

  • Affinity purification (e.g., pre-adsorption) from the source serum provides only limited specificity (background and cross-reactivity are common limitations of pAbs)
  • Animal variability and purification process can result in an unknown amount of lot variability

Excellent

  • Clones selected and combined in sets designed to eliminate off-target binding (i.e., cross-reactivity) in the tested applications
  • Recombinant technology helps ensure that every lot comprises exactly the same clones

Very good

  • Can select for a clone that has no off-target binding
  • Propagation of monoclonal cell line can result in clonal drift; sequence of antibody is not definitively known
Host and target species available
  • Goat anti-mouse (GAM)
  • Goat anti-rabbit (GAR)
  • Rabbit anti-mouse (RAM)
  • Rabbit anti-goat (RAG)
  • Many others
  • Goat anti-mouse (GAM)
  • Goat anti-rabbit (GAR)
  • Rabbit anti-mouse (RAM)
  • Rabbit anti-goat (RAG)

Traditional monoclonal antibody production uses mouse or rat as the host. Monoclonals are seldom used as secondary antibodies.