Antibody and Protein Conjugation


 

 

Conjugation Custom Services


Rockland has decades of experience in conjugation of antibody and protein reagents. Our conjugation experience spans from service to the research, pharmaceutical, diagnostics to biotech communities. Rockland produces a wide array of customizable, on-demand conjugation services to meet specific assay and reagent needs. Through our experience we have developed scalable processes from milligram (mg) to gram (g) quantities of conjugates.


 Fluorescent Dyes Chart

 

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1. Conjugated Antibodies

2. Colorimetric & Chemiluminescent Substrates

3. Application & Related Services

 


 

 

Conjugated Antibodies

 

 

 Horseradish Peroxidase (HRP) Conjugated Antibodies are the most popular antibody conjugates and can be used in Western Blot Analysis (immunoblotting), ELISA and Immunohistochemistry. In addition to the HRP-conjugated antibodies, Rockland produces secondary antibodies that are Fluorescein conjugated, as well as conjugates to Rhodamine, Texas Red, Phycoerythrin (PE) and dyes for multiplex platforms like the LI COR Odyssey Imaging System or Molecular Devices multi-mode readers. These dyes consist of Atto Dye and DyLight™ Dyes.

 

 

 

 Fluorescent antibodies can be used to analyze the distribution of proteins, glycoproteins and other antigen targets including small biological and non-biological molecules. This provides considerable insight into the movement and utilization of antibodies throughout various biological systems. Primary, or direct, immunofluorescence uses a single antibody conjugated directly to a fluorescent dye. Secondary, or indirect, immunofluorescence uses two antibodies; a primary antibody which recognizes the target biomolecule and binds to it and a secondary antibody conjugated to a fluorescent dye, which recognizes and binds to the primary antibody and indirectly localizes the target for detection.

 

 

 

 Rockland conjugates a broad group of secondary antibodies to many of the classic fluorescent markers including fluorescein, Texas Red, and Phycoerythrin. All of the conjugates are ideal for various immunofluorescence based assays including fluorescent western blotting, immunofluorescence microscopy, FLISA, and more.

 

 
fluorescent secondary antibody absorption / emission spectrums   Rockland also produces many next generation fluorochrome dyes designed for detection of primary antibodies in multiplex, multi-color analysis. Next generation fluorochrome conjugates (Atto-tec dyes, DyLight™ dyes) offer superior absorption (high extinction coefficient), high fluorescence quantum yield, and superior high photostability.

Excitation and Emission spectra of selected ATTO-TEC dyes available for the labeling of custom antibodies.

Click here for tips to select the correct secondary antibody.
 
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Colorimetric & Chemiluminescent Substrates

 
Reporter enzymes are used extensively in molecular biology to detect immune complexes. Horseradish Peroxidase (HRP) is a widely used reporter enzyme, and depending on the substrate, can yield a colorimetric, chromogenic or luminescent product. Alkaline phosphatase is also used, most typically as the reporter in chromogenic western blot assay format. 
 
Rockland provides extremely sensitive chemiluminescent HRP substrates which are nonradioactive and provide enhanced luminol-based chemiluminescent detection of horseradish peroxidase (HRP) in western blotting and enzyme immunoassay (EIA) use. Rockland also manufactures substrates with 3, 3', 5, 5' – Tetramethylbenzidine (TMB) to aid in the detection of custom antibody conjugates. 
 
All substrates are available in bulk.
  

 

ABTS   PICOMAX-110
TMBE-100   FEMTOMAX-110
TMBM-100    

 

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Applications and Related Services

 
Common applications and related services include ELISA, western blotting, FLOW cytometry, immunohistochemistry and immunofluorescent microscopy

 

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Protein conjugates can be made with the following:

  • Peroxidase (HRP)
  • BIOTIN
  • Alkaline Phosphatase
  • Glucose Oxidase
  • B-Galactosidase
  • Fluorescent dyes
    • Atto Dye
    • Fluorescein
    • DyLight Dye
    • Phycoerythrins
    • Rhodamine
    • Texas Red

     

         
 Fluorescent Buffers