Spheroid tissue stained using a Goat anti-Mouse IgG , Superclonal Recombinant Secondary Antibody conjugated to Alexa Fluor Plus 488

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. Our 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 in vitro manufactured using synthetic genes after the first immunization. Each Superclonal secondary antibody is formulated and optimized to help achieve excellent results in ELISA, cell and tissue imaging (ICC/IF and IHC) and flow cytometry applications.

Features of Superclonal secondary antibodies

  • Developed as recombinant monoclonal antibodies to enable precise and accurate detection
  • Formulated to recognize multiple epitopes (e.g., detection of both heavy and light chain of primary antibodies) of target antibodies
  • Selected and optimized for use with western blotting, ELISA, cell and tissue imaging (ICC/IF &IHC) and flow cytometry 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), alkaline phosphorate (AP) and selected Invitrogen Alexa Fluor and Alexa Fluor Plus dyes
 

Superclonal recombinant secondary antibodies demonstrate lot-to-lot consistency

HeLa cells stained to show densitometric analysis of 4 lots of recombinant antibodies’ consistency

Figure 1: Invitrogen Superclonal secondary antibodies are designed to provide lot-to-lot consistency. Endogenous TOMM20 in HeLa cells were labeled with rabbit TOMM20 monoclonal primary antibody, which was then detected with four different lots of Goat anti-Rabbit IgG (Heavy Chain), Superclonal Recombinant Secondary Antibody, Alexa Fluor Plus 488 (Cat. No. A55053) (green). Comparable signal and noise fluorescence intensities from these four lots demonstrate lot-to-lot consistency of Invitrogen Superclonal secondary antibodies. Inset represents control cells with no primary (NP) antibody to assess noise.

Superclonal recombinant secondary antibodies help enable precise and accurate detection

HeLa cells stained to show morphological and protein targets using Superclonal secondary antibodies in ICC

Figure 2: Invitrogen Superclonal secondary antibodies are designed to provide enhanced specificity. Endogenous HDAC2 in HeLa cells were labeled with mouse HDAC2 Monoclonal primary antibody, which was then detected with Goat anti-Mouse IgG (H+L), Superclonal Recombinant Secondary Antibody, Alexa Fluor Plus 647 (Cat. No. A55060) (red). Simultaneously, LRP130 was labeled with rabbit LRP130 antibody followed by anti-rabbit secondary antibody conjugated to Alexa Fluor Plus 488 (green). Goat anti-Mouse IgG (H+L), Superclonal Recombinant Secondary Antibody, Alexa Fluor Plus 647 (Cat. No. A55060) specifically detects mouse primary antibody and does not cross-react with rabbit primary antibody.

Compare the technology

Superclonal antibodies compared to traditional polyclonal antibodies

 Traditional polyclonal antibodies
(pAb)		Superclonal secondary antibodies
How they are madeAffinity purification (by positive and/or negative selection) of antibodies from the serum of immunized animals.Produce and characterize numerous individual recombinant antibodies, then carefully screen, select, and pool specific ones to achieve enhanced performance.
What they areA large, undefined pool of antibodies from the host serum, selected by affinity purification. Contain multiple epitopes, the exact numbers of which are generally undetermined.Precisely characterized sets of specific recombinant antibodies having known, complementary sets of epitopes and affinity binding features.
Epitope coverage and signal amplification

Excellent

Broad epitope coverage for the target antibody ensures good sensitivity and signal amplification.

Excellent

Individual recombinant secondary antibodies are selected and pooled to provide complementary, specific and optimal epitope coverage and binding quality to maximize performance in key applications.

Specificity

Based on the individual host animal used for immunization, specificity of pAb can differ vastly. Affinity purification (e.g., pre-adsorption) of pAb from the source serum provides some additional improvement of specificity.

Individual clones and the final pool of clones are screened (using positive and/or negative selection) to eliminate cross reactivity and ensure high specificity in key applications.
Lot-to-lot consistency
Animal variability and purification process can result in variability between lots.
Recombinant antibody technology ensures high lot-to-lot consistency.