Colorimetric and Chemiluminescent Assays

Colorimetric and Chemiluminescent Assays


Rockland offers antibody and protein conjugation services associated with a wide range of common reporter molecules used in a 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 ßgal); in addition to biotin, digoxigenin and assorted haptens.








TrueBlot Mouse

TrueBlot Rabbit HRP

Anti-Mouse IgG (H&L)  

(Goat) Antibody

 Mouse TrueBlot® ULTRA

Anti-Mouse Ig HRP

Rabbit TrueBlot® 

 Anti-Rabbit IgG HRP

 HRP Conjugated                 Bioting Conjugated                   ALP Conjugated  

HRP fig


Horseradish Peroxidase (HRP) Conjugated Secondary Antibodies are the most popular antibody conjugates and are widely used to obtain chemiluminescent results in western blotting or other immunoblotting, ELISA, and immunochemistry. HRP is a heme-containing enzyme that catalyses the oxidation of luminol to 3-aminophtalate via serveral intermediates causing a low-intensity light emission at 428 nm. In the presence of certain chemicals (enhanced chemiluminescence or ECL), the light emitted is enhaced up to 1000-fold. The intensity of light of light is a measure of the number of enzyme molecules reacting.




TrueBlot®: Clear Detection of Blotted Target Bands




Rockland's TrueBlot® product line enables researchers to generate publication-quality IP western blot data. Simply substitute your conventional HRP blotting reagent with Goat TrueBlot®, Mouse TrueBlot®, Rabbit TrueBlot®, and Sheep TrueBlot®. TrueBlot® is a horseradish peroxidase conjugated immunoblotting detection reagent, which enables unhindered detection of blotted target bands. This results in unparalleled clarity in results and high quality imaging suitable for reproduction. 




Biotin Conjugated Antibodies


biotin fig



Biotin conjugates are typically used as an intermediary in immunological assays. Biotin has a high affinity and specificity for binding to avidin or streptavidin and is regularly used in ELISA, Western blotting, and affinity chromatography. Biotin is a water-soulable B-vitamin (B7) also called vitamin H. In addition to its importance in health and diet it is widely used in research to conjugate proteins for biochemical assays, protein purification or separation. Due to the small size of biotin (MW = 244.31 g/mol), biotinylation will unlikely hinder the natural function of the molecule. 






Alkaline Phosphatase Conjugated Antibodies




Alkaline phosphatase (ALP) is a hydrolase enzyme that dephosphorylates nucleotides, protein and alkaloids in preferably alkaline environments. The enzyme is derived from calf intestine, and is most commonly used in immunoassays that result in chromogenic detection. With the addition of ALP substrates, ALP conjugates produce a color change that can be easily read and quantified at 405nm.




Colorimetric & Chemiluminescent Substrates




Reporter enzymes are used extensively in molecular biology and biochemistry 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 ECL 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.




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