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Pierce™ Fab Micro Preparation Kit Thermo Scientific™

Thermo Scientific Pierce Fab Micro Preparation Kit uses immobilized papain protease to digest human or mouse IgG antibodies to make separate Fab and Fc fragments and subsequently to purify the Fab using Protein A agarose.

Features of the Fab Micro Preparation Kit:

High capacity—use the micro kit for 25 to 250 µg IgG.
Enzyme-free digestion products—Immobilized Papain (beaded agarose resin) provides for control of the digestion reaction and complete removal of resulting antibody fragments from the proteolytic enzyme
Suitable for human and other species of IgG—the kit procedure is optimized for human, mouse and rabbit IgG. Papain-based digestion is effective for certain other species and subclasses of IgG (e.g., goat and pig), although the purification step requires that the antibody effectively binds to Protein A. (Note: for best results with mouse IgG1, use Part No. 44980.)
Provides ready-to-use Fab—digestion and final recovery of purified Fab fragments occurs in neutral pH sodium phosphate buffer, suitable for storage or immediate use in typical applications
Complete—kits include all reagents needed to prepare and purify antibody fragments
Fast—spin format greatly reduces sample processing time
Flexible—Protocols are included for multiple species and IgG subclasses, as well as sample size and concentration.
Efficient—enhanced yield and sample purity

These Fab Preparation Kits are suitable for human, rabbit, mouse and other species and subclasses of IgG. The papain antibody digestion reaction is performed in convenient disposable spin columns that allow efficient removal of the immobilized protease and maximum recovery of the IgG fragments. Also included in the kits are Thermo Scientific NAb Protein A Spin Columns and buffers to efficiently purify the resulting fragments. Protein A binds the Fc fragments and undigested IgG, allowing the pure Fab fragments to be recovered in the flow-through fraction. The kits also include Thermo Scientific Zeba Desalting Spin Columns for preparing the IgG sample quickly without dilution instead of utilizing time-consuming dialysis steps.

The kits use papain, a nonspecific thiol-endopeptidase, to enzymatically cleave whole IgG just above the hinge region to create two separate Fab fragments and one Fc fragment per antibody molecule. Because the papain protease is supplied in immobilized form as beaded agarose resin, the digestion reaction is easily stopped by removing the resin from the IgG solution; the result is digest products that are enzyme-free.

Related Products
Pierce™ Fab Preparation Kit
Pierce™ F(ab')2 Micro Preparation Kit
Pierce™ Mouse IgG1 Fab and F(ab')2 Micro Preparation Kit

HT ExoSAP-IT™ Fast High-Throughput PCR Product Cleanup Applied Biosystems™

HT ExoSAP-IT Fast High-Throughput PCR Product Cleanup, with its shortened 14 minute protocol, is a highly stable, customized formulation of the original ExoSAP-IT reagent. It is ideal for high-throughput PCR product cleanup using liquid handling platforms and multi-channel pipettes. Multiple packaging formats (vials, 8-tube strips, 96-well plates) ensure ease of use and flexibility in experimental design.

Like the original formulation, HT ExoSAP-IT Fast reagent is a mixture of Exonuclease I and Shrimp Alkaline Phosphatase for the enzymatic removal of excess primers and dNTPs following a PCR reaction. This enzymatic cleanup reagent has a decreased viscosity yet maintains the same convenience and stability that you have come to expect from the ExoSAP-IT product line. As with all products in the ExoSAP-IT family, HT ExoSAP-IT Fast reagent allows PCR cleanup with a single pipetting step. .

HT ExoSAP-IT Fast reagent is scalable for PCR cleanup from a single tube up to a 384-well plate PCR reaction.
•14 minute protocol is half the time of standard protocols
•One-tube PCR cleanup — add HT ExoSAP-IT Fast reagent directly to PCR product
•100% sample recovery — no loss of PCR products regardless of the fragment size
•Stable at 4°C for one month
•Removes excess primers and dNTPs — does not interfere with downstream applications
   -sequencing
   -SNP analysis
   -single base extension
   -fragment analysis

Adding to our family of patented ExoSAP-IT products, HT ExoSAP-IT Fast High-Throughput PCR Cleanup reagent quickly provides accurate and consistent results in high-throughput applications. HT ExoSAP-IT Fast reagent is over 50% faster than standard HT ExoSAP-IT reagent with a 14 minute total protocol time.
•Add it directly to the PCR product and incubate at 37°C for just 7 minutes.
• After PCR treatment, inactivate it by simply heating to 80°C for another 7 minutes.

This product is ideal for high volume labs that require quick sample turn around. With such a brief and simple protocol, proper sample cleanup never needs to be sacrificed due to processing or instrument time constraints. Ensure the highest quality Sanger sequencing results while keeping sample processing moving quickly and efficiently.

PCR products cleaned up with HT ExoSAP-IT Fast reagent prior to automated sequencing displayed superior sequencing results compared to the untreated equivalent (Fig. 2).

HT ExoSAP-IT Fast reagent displayed excellent stability in all formats tested including vials, 8-tube strips, and 96-well plates. The product showed no loss in function after 10 freeze thaw cycles (Fig. 3), and is stable at variable temperatures for extended periods of time compared to enzyme mix equivalents. With excellent stability and a greater than 50% faster protocol, HT ExoSAP-IT Fast reagent is ideal for high-throughput assays where robotic platforms are utilized and large volumes of samples are processed in successive testing runs.

Collectively, these results clearly demonstrate the effectiveness of HT ExoSAP-IT Fast reagent to ensure the highest quality sequences while keeping sample processing moving quickly and efficiently.

Staphylococcus aureus V-8 Protease Thermo Scientific™

Thermo Scientific Staphylococcus aureus V-8 Protease also known as endoproteinase Glu-C, has two pH optima, one at pH 4.0 and the other at pH 7.8, and is specific for cleavage at the carboxy terminus of glutamic acid and aspartic acid residues. The protease is specific only for glutamic digestion in buffers which do not contain phosphate at either pH optimum. Phosphate buffers at pH 7.8 will facilitate digestion at both glutamic and aspartic acid residues.

Features of Staphylococcus aureus V-8 Protease:

Specificity—Specificity for glutamic acid is achieved in ammonium bicarbonate, pH 7.8 or ammonium acetate, pH 4
Activity—Enzyme is active in the presence of many denaturing agents such as SDS, urea and guanidine-HCl

S. aureus V-8 protease protease is very stable, being able to retain its activity even in 6 M urea, 5.5 M guanidine·HCl, 0.5% SDS, or 0.5% SDS + 6 M urea at 0°C, pH 7.6. This is related to the fact that the native structure of the protease is stabilized mainly by electrostatic interactions, and not by hydrogen bonding and β-structures.

There are two major problems with any proteolytic digestion. The first problem is that the reaction must be quenched to stop the digestion, thus diluting out the sample. The second problem is that the sample is contaminated with the protease and possibly autodigested protease fragments. These problems are surmounted with immobilized V-8 protease. With immobilized proteases, no quenching is necessary and there are no problems with protease contamination of the sample or autodigestion of the protease. Simply separate the sample solution from the agarose matrix and these problems are eliminated.

Related Products
Immobilized V-8 Protease Kit

HT ExoSAP-IT™ Fast High-Throughput PCR Product Cleanup Applied Biosystems™

HT ExoSAP-IT Fast High-Throughput PCR Product Cleanup, with its shortened 14 minute protocol, is a highly stable, customized formulation of the original ExoSAP-IT reagent. It is ideal for high-throughput PCR product cleanup using liquid handling platforms and multi-channel pipettes. Multiple packaging formats (vials, 8-tube strips, 96-well plates) ensure ease of use and flexibility in experimental design.

Like the original formulation, HT ExoSAP-IT Fast reagent is a mixture of Exonuclease I and Shrimp Alkaline Phosphatase for the enzymatic removal of excess primers and dNTPs following a PCR reaction. This enzymatic cleanup reagent has a decreased viscosity yet maintains the same convenience and stability that you have come to expect from the ExoSAP-IT product line. As with all products in the ExoSAP-IT family, HT ExoSAP-IT Fast reagent allows PCR cleanup with a single pipetting step. .

HT ExoSAP-IT Fast reagent is scalable for PCR cleanup from a single tube up to a 384-well plate PCR reaction.
•14 minute protocol is half the time of standard protocols
•One-tube PCR cleanup — add HT ExoSAP-IT Fast reagent directly to PCR product
•100% sample recovery — no loss of PCR products regardless of the fragment size
•Stable at 4°C for one month
•Removes excess primers and dNTPs — does not interfere with downstream applications
   -sequencing
   -SNP analysis
   -single base extension
   -fragment analysis

Adding to our family of patented ExoSAP-IT products, HT ExoSAP-IT Fast High-Throughput PCR Cleanup reagent quickly provides accurate and consistent results in high-throughput applications. HT ExoSAP-IT Fast reagent is over 50% faster than standard HT ExoSAP-IT reagent with a 14 minute total protocol time.
•Add it directly to the PCR product and incubate at 37°C for just 7 minutes.
• After PCR treatment, inactivate it by simply heating to 80°C for another 7 minutes.

This product is ideal for high volume labs that require quick sample turn around. With such a brief and simple protocol, proper sample cleanup never needs to be sacrificed due to processing or instrument time constraints. Ensure the highest quality Sanger sequencing results while keeping sample processing moving quickly and efficiently.

PCR products cleaned up with HT ExoSAP-IT Fast reagent prior to automated sequencing displayed superior sequencing results compared to the untreated equivalent (Fig. 2).

HT ExoSAP-IT Fast reagent displayed excellent stability in all formats tested including vials, 8-tube strips, and 96-well plates. The product showed no loss in function after 10 freeze thaw cycles (Fig. 3), and is stable at variable temperatures for extended periods of time compared to enzyme mix equivalents. With excellent stability and a greater than 50% faster protocol, HT ExoSAP-IT Fast reagent is ideal for high-throughput assays where robotic platforms are utilized and large volumes of samples are processed in successive testing runs.

Collectively, these results clearly demonstrate the effectiveness of HT ExoSAP-IT Fast reagent to ensure the highest quality sequences while keeping sample processing moving quickly and efficiently.

Human Cytosol Gibco™

Pooled human liver cytosolic fractions.

Total protein concentration: 20 mg/ml

We provide liver subcellular fractions from a variety of tox species, including human, nonhuman primates (Cynomolgus Monkey and Rhesus Monkey), dog (Beagle), rat (Sprague-Dawley), mouse (CD-1 and Balb-c), and trout (Oncorhynchus mykiss). Custom species are available upon request.

For research use only. Not intended for human or animal therapeutic or diagnostic use.

Related Link

Learn more about our microsomes and subcellular fractions

Vivid™ Regeneration System, 100X

Vivid® Regeneration System, 100X, is a Vivid® CYP450 Screening Kit component consisting of glucose-6-phosphate and glucose-6-phosphate dehydrogenase, which converts NADP+ into NADPH, which is required to start the CYP450 reaction. Vivid® Regeneration System, 100X, is included in the Vivid® Screening Kits, but may also be purchased separately.

More About Vivid® CYP450 Screening Kits
Vivid® CYP450 Screening Kits are high-throughput fluorescence-based assays for detection of enzyme-drug interactions and CYP450 inhibition. Vivid® CYP450 Screening Kits include Vivid® Substrate, Vivid® Fluorescent Standard, reaction buffer, Vivid® Regeneration System, NADP+, and CYP450 BACULOSOMES® Plus Reagent. Vivid® Fluorogenic Substrates are blocked dyes that yield minimal fluorescence signal until cleaved or hydroxylated. Oxidation at either of two potential sites releases the highly fluorescent product. CYP450 BACULOSOMES® Plus Reagents are microsomes prepared from insect cells expressing a human CYP450 isozyme and human cytochrome P450 reductase. CYP450 BACULOSOMES® Plus Reagents offer a distinct advantage over human liver microsomes in that only one CYP450 isozyme is expressed, thereby preventing metabolism by other CYP450s.

>> View all Vivid® CYP450 Screening Kits

For Research Use Only. Not for any animal or human therapeutic or diagnostic use.

Sphingosine Kinase 1a Recombinant Protein Invitrogen™

Mouse SPHK1a Recombinant Protein for Ctrl

PCR Product Pre-Sequencing Kit Applied Biosystems™

The PCR Product Pre-Sequencing Kit uses a novel, enzymatic clean-up method to pre-treat PCR products prior to sequencing, without any subsequent purification or separation steps. The two hydrolytic enzymes used in this kit, shrimp alkaline phosphatase and exonuclease I, effectively remove excess dNTPs and primers present in the final PCR product reaction mixture. The enzymes are conveniently added to an aliquot of the PCR product mixture, incubated at 37°C, and then inactivated at 80°C. The result is a cleaner PCR product, free from excess nucleotides and primers, and ready to be sequenced with standard sequencing reagents.

Vivid™ NADP+ , 10mM

Vivid® NADP+, 10 mM, is a Vivid® CYP450 Screening Kit component consisting of 10 mM NADP+ in 100 mM potassium phosphate, pH 8.0. Vivid® NADP+, 10 mM, is included in the Vivid® screening kits, but may also be purchased separately.

More About Vivid® CYP450 Screening Kits
Vivid® CYP450 Screening Kits are high-throughput fluorescence-based assays for detection of enzyme-drug interactions and CYP450 inhibition. Vivid® CYP450 Screening Kits include Vivid® Substrate, Vivid® Fluorescent Standard, reaction buffer, Vivid® Regeneration System, NADP+, and CYP450 BACULOSOMES® Plus Reagent. Vivid® Fluorogenic Substrates are blocked dyes that yield minimal fluorescence signal until cleaved or hydroxylated. Oxidation at either of two potential sites releases the highly fluorescent product. CYP450 BACULOSOMES® Plus Reagents are microsomes prepared from insect cells expressing a human CYP450 isozyme and human cytochrome P450 reductase. CYP450 BACULOSOMES® Plus Reagents offer a distinct advantage over human liver microsomes in that only one CYP450 isozyme is expressed, thereby preventing metabolism by other CYP450s.

>> View all Vivid® CYP450 Screening Kits

For Research Use Only. Not for any animal or human therapeutic or diagnostic use.

Pierce™ F(ab')2 Micro Preparation Kit Thermo Scientific™

The Thermo Scientific Pierce F(ab')2 Micro Preparation Kit uses immobilized pepsin protease to selectively digest whole human and other IgG antibodies to make F(ab')2 fragments that retain antigen binding activity.

Features of the F(ab')2 Micro Preparation Kit:

Ideal when sample size is limited—Use the micro kit for 25 to 250 µg IgG.
Enzyme-free digestion products—Immobilized Pepsin (beaded agarose resin) provides for control of the digestion reaction and complete removal of resulting antibody fragments from the proteolytic enzyme
Suitable for human and other species of IgG—the kit procedure is optimized for human, rabbit and mouse IgG. Pepsin-based digestion is effective for other species and subclasses of IgG, although the purification step requires that the antibody effectively binds to Protein A. (Note: for best results with mouse IgG1, use Part No. 44980.)
Complete—kits include all reagents needed to prepare and purify antibody fragments
Fast—spin format greatly reduces sample processing time
Flexible—protocols are included for multiple species and IgG subclasses, as well as sample size and concentration.
Efficient—enhanced yield and sample purity

These F(ab')2 Preparation Kits are suitable for human, rabbit, mouse and other species and subclasses of IgG except mouse IgG1. The antibody digestion reaction with pepsin agarose is performed in convenient disposable spin columns that allow efficient removal of the immobilized protease and maximum recovery of the IgG fragments. Also included in the kits are Thermo Scientific NAb Protein A Spin Columns and buffers to efficiently purify the resulting fragments. Protein A binds the Fc fragments and undigested IgG, allowing the pure F(ab')2 fragments to be recovered in the flow-through fraction. The kits also include Thermo Scientific Zeba Desalting Spin Columns for preparing the IgG sample quickly without dilution instead of utilizing time-consuming dialysis steps.

The kits use pepsin, a nonspecific endopeptidase, to enzymatically digest the Fc portion of whole IgG to yield the fragment known as F(ab')2. This fragment is composed of a pair of Fab' units connected by two disulfide bonds. Because the pepsin protease is supplied in immobilized form as beaded agarose resin, the digestion reaction is easily stopped by removing the resin from the IgG solution; the result is digest products that are enzyme-free.

Related Products
Pierce™ F(ab')2 Preparation Kit
Pierce™ Fab Micro Preparation Kit
Pierce™ Mouse IgG1 Fab and F(ab')2 Micro Preparation Kit

PEST Recombinant Protein Invitrogen™

Human PTPN12 Recombinant Protein for Ctrl

Human microsomes, single donor

Human microsomes, single donor

Immobilized Trypsin, TPCK Treated (Agarose Resin) Thermo Scientific™

Thermo Scientific Pierce Immobilized TPCK Trypsin is a serine endoprotease that is applicable to amino acid analysis and protein sequencing, mapping and structural studies; the immobilized form allows sample separation after treatment.

Features of Thermo Scientific Pierce Immobilized TPCK Trypsin

• Cleaves at carboxyl side of arginine and lysine residues
• Digestion conditions: pH 7.5-9.0, 37°
• Immobilized for ease of separation from cleavage products or affinity purification of Trypsin Inhibitors

Trypsin has a wide range of applications including amino acid analysis and protein sequencing, mapping and structural studies. Enzymes such as trypsin and chymotrypsin have become important tools in sequencing studies since they are highly selective in their cleavage of peptide bonds. Trypsin cleaves only those peptide bonds in which the carboxyl group is contributed by a lysine or an arginine residue, regardless of the length or amino acid sequence of the chain. Our Immobilized TPCK Trypsin can be substituted for free trypsin in many application and is advantageous because it minimizes autolysis, eliminates contamination of a sample with the protease and allows control of the digestion by removing the trypsin. Immobilized trypsin is also more stable against heat-induced denaturation, resulting in longer maintenance of activity.

Applications:
• Removal of adherent cells from tissue culture flasks
• Preparing tryptic fragments for Edman degradation sequencing
• Immobilized trypsin can be used to purify soybean trypsin inhibitor
• Use our Pierce Trypsin, MS Grade (Part No. 90057) for mass spectrometry workflows

Trypsin is a 23.8kDa pancreatic serine endoprotease derived from trypsinogen, an inactive precursor zymogen, after enzymatic removal of an n-terminal leader sequence by enterokinase. Once some trypsin has been formed it can catalyze the conversion of more trypsinogen into its catalytically-active form. Trypsin is treated with L-1-tosylamido-2-phenylethyl chloromethyl ketone (TPCK) to inhibit contaminating chymotrypsin activity without affecting trypsin activity.

Related Products
Trypsin, TPCK Treated

CYP2C19 BACULOSOMES™ Plus Reagent, rHuman

Cytochrome P450 BACULOSOMES® Plus Reagents are microsomes prepared from insect cells infected with recombinant baculovirus containing a human CYP450 isozyme, as well as human cytochrome P450 reductase. For this particular isozyme (CYP2C19), human cytochrome b5 is also included.
• Single human P450 isozyme for detailed drug metabolism studies
• High specific activity increases dynamic range and sensitivity
• No background metabolism for clear results

Single Overexpressed Human CYP450 Isozyme
CYP450 BACULOSOMES® Plus Reagents offer a distinct advantage over Human Liver Microsomes (HLMs) in that only one CYP450 isozyme is expressed, thereby preventing metabolism by other CYP450s or other classes of drug metabolizing enzymes.

High Activity Results in Increased Sensitivity
Compound rankings based on metabolism and inhibition profiles observed with CYP450 BACULOSOMES® Plus Reagents are very similar to those seen with HLMs for most compounds tested. However, CYP450 BACULOSOMES® Plus Reagents activity and metabolic rates with most substrates is significantly higher than those seen with HLMs. This results in a broad dynamic range and high sensitivity in assays utilizing CYP450 BACULOSOMES® Plus Reagents. The high level of activity and reproducibility of the enzyme component observed in the reaction with most probe substrates makes CYP450 BACULOSOMES® Plus Reagents well suited to high-throughput screening formats.

High Signal to Noise Ratio
No background metabolism by endogenous insect CYP450s has been detected with any of the Vivid® probe substrates tested so far.

Applications: isozyme identification, DMPK analysis, isozyme-specific metabolism, and inhibition screening

For Research Use Only. Not for any animal or human therapeutic or diagnostic use.

Human MMP2 Recombinant Protein Invitrogen™

Human MMP2 Recombinant Protein for Ctrl
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