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        Enzymes


 

Enzymes


Rockland manufactures a variety of enzymes for use in manufacturing and assay development.

 

Process of Enzyme Activity


 enzymesteps

 

 


 

Rockland offers several different types of enzymes

Nucleases

Collagenases

Kinases

Conjugates & Substrates

  


  

Nucleases


Nucleases are enzymes that cleave the phosphodiester bonds that connect nucleotides. This class of hydrolases allow for study and manipulation of nucleic acids, providing a platform for genetic recombination and sequencing assays.


           Nuclease S1         

Micrococcal Nuclease S7

 


 

Kinases 

 


Rockland offers its customers a highly purified Polynucleotide Kinase T4 clone, isolated from Escherichia coli. This enzyme is commonly used to label DNA or RNA through the process of phosphorylation. Polynucleotide Kinase T4 will catalyze the transfer of the terminal phosphate ion of ATP molecules to the 5' hydroxyl terminus of polynucleotides, such as DNA and RNA, or oligonucleotides.

 

Polynucleotide Kinase T4

 

 


 

Collagenase 


Collagenases(link to Collagenase Page) are a family of enzymes that hydrolyze the peptide bonds found in collagen. Breaking these bonds in extracellular matrices, built from collagen, is the first step in tissue dissociation and isolation. Collagenases can be used for:

  • Tissue isolation:  of adipocytes, hepatocytes, cells from lung, epithelium, adrenal tissue, cardiomyocytes, cells from bone, cartilage, muscle, thyroid, endothelium, mammary, human and porcine pancreatic islet cells.
  • Tissue treatment to provide gentle, selective digestion of the intercellular matrix with little damage to cells or loss of viability.

 


 

Conjugates and Substrates 


Rockland offers antibody and protein conjugates with a wide range of common enzymatic reporter molecules used in a molecular immunology. The reporter molecules, also called antibody labels, typically include enzymes such as horseradish peroxidase (HRP), alkaline phosphatase (ALP), glucose oxidase (GO) and beta galactosidase (BGAL or ßgal).
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 chemiluminescent 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.


For each reporter enzyme conjugate, Rockland offers a variety of substrates that will generate either a soluble or insoluble colored product.  Soluble substrates are useful to quantitate analytes in liquid phase assays like ELISA.  Insoluble substrates are excellent to localize analytes in solid phase assays like western blotting (WB) and immunohistochemistry (IHC).  Rockland’s FemtoMax™ and PicoMax™ chemiluminescent substrates are available for horseradish peroxidase for ELISA and Western Blots.

Enzymatic Processes


Rockland’s experience in working with antibodies allows us to process immunoglobulins into their constituent components. In some assays, it is preferable to use only the antigen-binding (Fab) portion of the antibody to prevent the constant (Fc) portion from binding to some cell surface receptors. For these applications, antibodies may be enzymatically digested using the enzymes: papain or pepsin to produce a Fab, Fc or F(ab')2 fragment of the antibody. The enzyme papain can be used to cleave a single immunoglobulin molecule into two Fab fragments and an Fc fragment. The enzyme pepsin cleaves below the antibody’s hinge region, so a F(ab')2 fragment and a Fc fragment is formed. Rockland offers a variety of labeled and unlabeled immunoglobulin fragments in addition to high-quality antibody fragmentation services that will provide you with optimum yields and predictable results for your custom F(ab’)2, Fab and Fc fragments.

 

New Lab Set Up 

 

 

 

Akt Cell Signaling Pathways