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        Custom Antibody Conjugation


Conjugation Overview

 

Rockland produces a wide array of ready-made conjugates, but also provides customizable, on-demand conjugation services for specific research needs. We produce antibody conjugates for both primary and secondary antibodies, while monoclonal and polyclonal antibodies are conjugated to Peroxidase (HRP), BIOTIN, alkaline phosphatase, and fluorescent dyes such as Atto Dye, Fluorescein, and DyLight Dye. Common applications include ELISA, western blotting, FLOW cytometry, immunohistochemistry and immunofluorescent microscopy.

  

      

 
 
 
 

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Conjugated Antibodies for Multiplex Systems

 

Antibodies for ELISA

ATTO Dye Conjugated Antibodies

 

Conjugated Antibodies for Multiplex Systems  

 

Antibodies for ELISA 

 

 ATTO Dye Conjugated Antibodies

 


 

Rockland offers antibody and protein conjugation services associated with a wide range of common reporter molecules used in molecular immunology. Reporter molecules, also called antibody labels or antibody tags, typically include enzymes such as horseradish peroxidase (HRP), alkaline phosphatase (ALP), glucose oxidase (GO) and beta galactosidase (BGAL or ß-gal); or fluorochromes including Fluorescein, DyLight dyes, phycoerythrins, Rhodamine, Texas Red and others along with biotin, digoxigenin and assorted haptens.

 

 

Spectra Chart of Selected Fluorescent Labels

 

Fluorescent label

 

 

Color

 

 

Abs (nm)

 

 

Em (nm)

 

 

MW (daltons)

 

DyLight™ 405

Violet

400

420

793

Aminomethylcoumarin (AMCA)

Violet Blue

353

442

410

ATTO 425

Blue

436

484

498

Cy2™

Blue Green

489

505

897

DyLight™ 488

Blue Green

493

518

1,011

ATTO 488

Green

501

523

981

Fluorescein (Fluorescein)

Green

495

528

390

ATTO 532

Yellow Green

532

553

1081

Cy3™

Yellow Green

552

565

949

DyLight™ 549

Yellow Green

550

568

982

Rhodamine (TRITC)

Orange

550

570

444

R-Phycoerythrin (RPE)

Orange

488

575

240,000

ATTO 550

Orange

554

576

791

Cy3.5™

Orange Red

581

596

1,286

Texas Red®

Red

596

620

625

ATTO 594

Red

601

627

1389

Allophycocyanin

Far-Red

650

660

100,000

Cy5™

Far-Red

650

667

975

ATTO 647N

Far-Red

644

669

843

DyLight™ 649

Far-Red

646

674

1,008

ATTO 655

Far-Red

663

684

887

Cy5.5™

Near Infra-Red

678

703

1,312

DyLight™ 680

Near Infra-Red

682

715

950

DyLight™ 800

Infra-Red

770

794

1,050

 

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       Peroxidase ( HRP ) Conjugated Secondary Antibody

Conjugated Secondary Antibodies 

Horseradish Peroxidase (HRP) Conjugated Secondary 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 secondary 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.

 

 

 

 

View All Rhodamine and Texas Red Conjugates               View All Phycoerythrin Conjugates


 
  

  

  

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 for the 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. 

 


  

Fluorescent Antibody Conjugates 

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. 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 ofselected ATTO-TEC dyes available for the labeling of custom antibodies.
 

 fluorescent secondary antibody absorption / emission spectrums  

 

 


  

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