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Genomics and Proteomics


Immunoassay Kit Format


Target Organism Class


eBioscience™ Mouse Hematopoietic Lineage Biotin Panel Invitrogen™

The eBioscience Mouse Hematopoietic Lineage Flow Panel contains 5 biotinylated antibodies that can be used to identify, enrich and/or deplete cells committed to the T, B, NK, myeloid and erythroid lineages based on their cell surface antigen expression.

Reactivity/Species
Mouse

Reported Application
Flow Cytometric Analysis

Active Rac1 Pull-Down and Detection Kit Thermo Scientific™

The Thermo Scientific Active Rac1 Pull-Down and Detection Kit is a complete kit for the selective enrichment and detection of GTP-bound Rac1 GTPase through specific protein interaction with the Pak1 protein-binding domain.

The Active Rac1 Pull-Down and Detection Kit includes purified GST-Pak1 protein-binding domain (PBD), glutathione agarose resin, positive and negative controls (GTPγS and GDP, respectively), lysis/binding/wash buffer, anti-Rac1 primary antibody, SDS sample buffer, spin columns and collection tubes. The kit was validated using lysates from NIH3T3 cells, a cell line that is known to have robust Rac1 activity.

Features of the Active Rac1 Pull-Down and Detection Kit:

Highly sensitive and accurate—optimized reagents, specific anti-Rac1 antibody and Western blot procedure ensure accurate controls and semi-quantitative results
Validated—functionally tested for Rac1 detection to ensure quality and performance
Compatible—effective with a variety of cell types from mouse, rat and human sources

Applications:
• Follow activation of Rac1 GTPase during cell differentiation, migration, division and cytoskeletal rearrangement
• Study Rac1 dependent lamellipodia formation
• Study the role of active Rac1 in cancer and angiogenesis
• Monitor Rac1 activity after stimulation with growth factors
• Screen small molecule inhibitors for their effect on Rac1 activity

The Active Rac1 Pull-Down and Detection Kit was validated for function and specificity of the active Rac1 enrichment method using cell lysates treated with GTPγS to activate endogenous Rac1 and compared to lysates treated with GDP to inactivate the small GTPase. GTPγS treatment traps Rac1 in the GTP-bound, active form, resulting in a strong signal when endogenous Rac1 is present. GDP treatment pushes Rac1 into the GDP-bound, inactive state, resulting in minimal or no signal, regardless of Rac1 protein levels. The kit is optimized for Western blot detection with an HRP-conjugated secondary antibody (Goat Anti-mouse IgG, Part No. 31430) and Thermo Scientific SuperSignal West Pico Chemiluminescent Substrate (Part No. 34080). The kit contains sufficient components for 30 pull-down assays.

Rac1 Background:
Rho family GTPases serve many cellular functions, including cell signaling, transcriptional regulation and organization of the actin cytoskeleton. This family of GTPases comprise Rho (RhoA, RhoB, and RhoC), Rac (Rac1, Rac2, Rac3, and RhoG), Cdc42 (Cdc42 and G25K), Rnd (Rnd1, Rnd2, and RhoE/Rnd3), RhoBTB family and the Miro family. These GTPases enable signal transduction from the plasma membrane to the cytosol through GPCR, tyrosine kinase, cytokine and adhesion receptors. Attachment to the plasma membrane is accomplished through geranylgeranyl lipid modifications at the carboxy-terminus of the protein. Rac1 activation results in actin polymerization and appears as membrane ruffling at the cellular periphery. Rac1 activation also results in lamelipodia formation.

The Rac1 GTPase transduces signals through tyrosine kinases, adhesion molecules or cytokine/chemokine receptors after stimulation with growth factors (EGF, insulin, PDGF, NGF), integrins (fibronectin) or chemoattractants (fMLP). For example, stimulation with EGF results in PI3 kinase activation, resulting in cell growth and reorganization at the cell periphery (membrane ruffling). Alternatively, tyrosine receptor kinase signaling through Rac1 leads to activation of the MAPK stress response pathways SAPK (JNK) and p38. Stimulation of cells with fibronectin results in integrin-mediated cell spreading. Two of the main effector proteins of Rac1 are Pak1 and phosphoinositol 4-phosphate 5-kinase. Pak1 (p65 Pak) is a kinase that activates the JNK pathway, while phosphoinositol 4-phosphate 5-kinase promotes actin filament assembly. Rac is critical for T-cell development and for promoting differentiating cells. However, the Rho family of GTPases can work agonistically during cell signaling and and antagonistically during differentiation.

More Product Data
Measure activation of small GTPases via their specific downstream effectors
Detection and localization of active GTPases in neuronal cell differentiation

FluoroMyelin™ Green Fluorescent Myelin Stain - Solution in Water Invitrogen™

The FluoroMyelin™ Green fluorescent myelin stain enables quick and selective labeling of myelin in brain cryosections in a single 20-minute labeling step plus washes. This stain can be used in conjunction with antibodies and other dyes, and with standard histochemical methods for cryosection material.

Isolectin GS-IB4 From Griffonia simplicifolia, Alexa Fluor™ 568 Conjugate Invitrogen™

Isolectin GS-IB4 is a 114,000-dalton glycoprotein that is part of a family of five tetrameric type I isolectins (IA4, IA3B, IA2B2, IAB3, and IB4) isolated from the seeds of the tropical African legume Griffonia simplicifolia. The A subunit prefers N-acetyl-D-galactosamine end groups while the B subunit is selective for terminal a-D-galactosyl residues. The red fluorescent Alexa Fluor® 568 isolectin GS-IB4 conjugate can be used in fluorescence microscopy.

View complete list of fluorescent dye-conjugated lectins ›

Wheat Germ Agglutinin, Alexa Fluor™ 555 Conjugate Invitrogen™

Wheat germ agglutinin (WGA) is one of the most widely used lectins in cell biology. Our Alexa Fluor® 555 conjugate of WGA exhibits the bright, orange fluorescence of the Alexa Fluor® 555 dye (excitation/emission maxima ~555/565 nm). Alexa Fluor® 555 WGA binds to sialic acid and N-acetylglucosaminyl residues.

View complete list of fluorescent dye-conjugated lectins ›

Active Cdc42 Pull-Down and Detection Kit Thermo Scientific™

The Thermo Scientific Active Cdc42 Pull-Down and Detection Kit is a complete kit for selective enrichment and detection of GTP-bound Cdc42 GTPase through specific protein interaction with the Pak1 protein-binding domain.

The Active Cdc42 Pull-Down and Detection Kit includes purified GST-Pak1 protein-binding domain (PBD), glutathione agarose resin, positive and negative controls (GTPγS and GDP, respectively), lysis/binding/wash buffer, anti-Cdc42 primary antibody, sample buffer, spin columns and collection tubes. The kit was validated using lysates from NIH 3T3 cells, a cell line that is known to have robust Cdc42 activity.

Features of the Active Cdc42 Pull-Down and Detection Kit:

Highly sensitive and accurate—optimized reagents, specific anti-Cdc42 antibody and Western blot procedure ensure accurate controls and semi-quantitative results
Validated—functionally tested for Cdc42 detection to ensure quality and performance
Compatible—effective with a variety of cell types from mouse, rat and human sources

Applications:
• Follow activation of Cdc42 GTPase during cell differentiation, migration, division, and cytoskeletal rearrangement
• Study the activation of Cdc42 during filopodia formation
• Monitor Cdc42 activity after stimulation with growth factors
• Screen small molecule inhibitors for their effects on Cdc42 activity

The Active Cdc42 Pull-Down and Detection Kit was validated for the function and specificity of the active Cdc42 enrichment method using cell lysates treated with GTPγS to activate endogenous Cdc42 and compared to lysates treated with GDP to inactivate the small GTPase. GTPγS treatment traps Cdc42 in the GTP-bound, active form, resulting in a strong signal when endogenous Cdc42 is present. GDP treatment pushes Cdc42 into the GDP-bound, inactive state, resulting in minimal or no signal regardless of Cdc42 protein levels. This kit is optimized for Western blot detection with an HRP-conjugated secondary antibody (Goat Anti-mouse IgG, Part No. 31430) and Thermo Scientific SuperSignal West Pico Chemiluminescent Substrate (Part No. 34080). The kit contains sufficient components for 30 pull-down assays.

Cdc42 Background:
Rho family GTPases serve many cellular functions, including cell signaling, transcriptional regulation and organization of the actin cytoskeleton. This family of GTPases comprise Rho (RhoA, RhoB, and RhoC), Rac (Rac1, Rac2, Rac3, and RhoG), Cdc42 (Cdc42 and G25K), Rnd (Rnd1, Rnd2, and RhoE/Rnd3), RhoBTB family and the Miro family. These GTPases enable signal transduction from the plasma membrane to the cytosol through GPCR, tyrosine kinase, cytokine and adhesion receptors. Attachment to the plasma membrane is accomplished through geranylgeranyl lipid modifications at the carboxy-terminus of the protein. Cdc42 activation results in the polymerization of actin filaments and filopodia formation.

The Cdc42 subfamily of RhoGTPase has been less well-characterized than the Rho and Rac families. Both Cdc42 and Rac1 are activated through tyrosine receptor kinase signaling leading to SAPK and p38 stress kinase pathway activation. Cdc42 is also activated by the chemoattractant fMLP in neutrophils. Fibronectin activates Cdc42 and Rac1 to induce cell spreading, and stimulation with TNF-alpha and IL-1 results in changes in the actin cytoskeleton. There is significant cross-talk between Cdc42 and Rac1, as they act in overlapping pathways, and in some cases, Cdc42 may act upstream of Rac1 during signal transduction. Some of the main effector proteins of Cdc42 are Pak1, N-WASP and IQGAP. Pak1 is a kinase involved in the activation of JNK in the SAPK stress pathway. N-WASP is an effector that induces filopodia formation, and IQGAP interacts with F-actin filaments. In differentiating neurons, Cdc42 plays an active role in neurite outgrowth. However, the Rho family of GTPases can work agonistically during cell signaling and antagonistically during differentiation.

More Product Data
Measure activation of small GTPases via their specific downstream effectors

Pierce™ BCA Protein Assay Reagent A Thermo Scientific™

BCA Protein Assay Reagent A is a component of the Pierce BCA Protein Assay Kit, a two-component, high-precision, detergent-compatible assay that is used for total protein concentration determination compared to a protein standard. Pierce BCA reagents provide accurate determination of protein concentration with most sample types encountered in protein research. The Pierce BCA assay can be used to assess yields in whole cell lysates, affinity-column fractions, purified proteins samples, as well as to monitor protein contamination in industrial applications. Compared to most dye-binding methods, the BCA assay is affected much less by protein compositional differences, providing greater concentration accuracy.

Compare all available BCA protein assays ›

Features of the Pierce BCA Protein Assay Kit (Reagents A and B) include:
Colorimetric—estimate visually or measure with a standard spectrophotometer or plate reader at 562 nm
Uniformity—exhibits less protein-to-protein variation than dye-binding methods (Bradford)
Compatibility—unaffected by typical concentrations of most ionic and non-ionic detergents
Assay time—30-min incubation; much easier and four times faster than classical Lowry methods
Assay range—linear working range for BSA of 20 to 2000 µg/mL
Sensitivity—detect down to 5 µg/mL with the enhanced protocol

How the assay works
The BCA Protein Assay combines the well-known reduction of Cu2+ to Cu1+ by protein in an alkaline medium with the highly sensitive and selective colorimetric detection of the cuprous cation (Cu1+) by bicinchoninic acid (BCA). The first step is the chelation of copper with protein in an alkaline environment to form a light blue complex. In this reaction, known as the biuret reaction, peptides containing three or more amino acid residues form a colored chelate complex with cupric ions in an alkaline environment containing sodium potassium tartrate.

In the second step of the color development reaction, BCA reacts with the reduced (cuprous) cation that was formed in step one. The intense purple-colored reaction product results from the chelation of two molecules of BCA with one cuprous ion. The BCA/copper complex is water soluble and exhibits a strong linear absorbance at 562 nm with increasing protein concentrations. The complex is approximately 100 times more sensitive (lower limit of detection) than the pale blue color of the first reaction.

The reaction is strongly influenced by four amino acid residues (cysteine, cystine, tyrosine, and tryptophan) in the amino acid sequence of the protein. However, unlike Coomassie dye-binding methods, the universal peptide backbone also contributes to color formation, helping to minimize variability caused by protein compositional differences.

Related products
Pierce BCA Protein Assay Kit
Pierce BCA Protein Assay Reagent B
Pierce BCA Solid

Vivid™ CYP2D6 Cyan Screening Kit

Vivid® CYP450 Screening Kits are high-throughput fluorescence-based assays for detection of enzyme-drug interactions and CYP450 inhibition. They provide an optimized method for studying isozyme-specific CYP450-drug interactions, metabolism, and inhibition. Vivid® CYP450 Screening Kits offer:
• Easy three-step procedure, 'mix and read' format, reactions at room temperature or 37°C
• Superior fluorescent properties and kinetics compared to conventional fluorogenic probes
• High signal-to-background ratio, broad dynamic range
• Compatible with multiple assay formats from 96-well to 1536-well

Simple Mix and Read Kit Format
Vivid® CYP450 Screening Kits include Vivid® Substrate, Vivid® Fluorescent Standard, reaction buffer, Vivid® Regeneration System, NADP+, and CYP450 BACULOSOMES® Plus Reagents. The CYP450 BACULOSOMES® Plus Reagents are microsomes prepared from insect cells expressing a human CYP450 isozyme (CYP2D6 in this case) 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.

Unique Vivid® Reagents for Bright Fluorescent Signals and Low Background
Vivid® 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 (Figure 1). They have superior fluorescence, solubility, and kinetic properties compared to conventional fluorogenic probes. This results in higher sensitivity, greater signal-to-noise ratio, and better assay reproducibility. Vivid® MOBFC Substrate is the substrate included in this particular kit. This substrate yields a product that emits a cyan fluorescence.

Flexible Assay Formats for Optimized Results
The sensitivity of the Vivid® CYP450 assays allows detection of weak inhibitors and miniaturization to as little as 2 µl per reaction. Assays may be set up in kinetic mode or in end-point mode to facilitate multi-plate screening (Figure 2). Assays may be performed at room temperature or 37°C.

Applications: high-throughput screening of enzyme-drug interactions, compound profiling for drug inhibition of cytochrome P450 isozymes, generation of predictive SAR models to guide compound acquisition

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

Wheat Germ Agglutinin, Alexa Fluor™ 594 Conjugate Invitrogen™

Wheat germ agglutinin (WGA) is one of the most widely used lectins in cell biology. Our Alexa Fluor® 594 conjugate of WGA exhibits the bright, red fluorescence of the Alexa Fluor® 594 dye (excitation/emission maxima ~590/617 nm). Alexa Fluor® 594 WGA binds to sialic acid and N-acetylglucosaminyl residues.

View complete list of fluorescent dye-conjugated lectins ›

FluoroMyelin™ Red Fluorescent Myelin Stain - Solution in Water Invitrogen™

The FluoroMyelin™ Red fluorescent myelin stain enables quick and selective labeling of myelin in brain cryosections in a single 20-minute labeling step plus washes. This stain can be used in conjunction with antibodies and other dyes, and with standard histochemical methods for cryosection material.

Isolectin GS-IB4 From Griffonia simplicifolia, Alexa Fluor™ 594 Conjugate Invitrogen™

Isolectin GS-IB4 is a 114,000-dalton glycoprotein that is part of a family of five tetrameric type I isolectins (IA4, IA3B, IA2B2, IAB3, and IB4) isolated from the seeds of the tropical African legume Griffonia simplicifolia. The A subunit prefers N-acetyl-D-galactosamine end groups while the B subunit is selective for terminal a-D-galactosyl residues. The red fluorescent Alexa Fluor® 594 isolectin GS-IB4 conjugate can be used in fluorescence microscopy.

View complete list of fluorescent dye-conjugated lectins ›

Concanavalin A, Tetramethylrhodamine Conjugate Invitrogen™

Concanavalin A (Con A) is one of the most widely used lectins in cell biology. Our tetramethylrhodamine conjugate of Con A exhibits the bright, orange-red fluorescence of tetramethylrhodamine (absorption/emission maxima ~555/580 nm). Tetramethylrhodamine Con A selectively binds to α-mannopyranosyl and α-glucopyranosyl residues.

View complete list of fluorescent dye-conjugated lectins ›

Wheat Germ Agglutinin, Alexa Fluor™ 680 Conjugate Invitrogen™

Wheat germ agglutinin (WGA) is one of the most widely used lectins in cell biology. Our Alexa Fluor® 680 conjugate of WGA exhibits the bright, red fluorescence of the Alexa Fluor® 680 dye (excitation/emission maxima ~679/702 nm). Alexa Fluor® 680 WGA binds to sialic acid and N-acetylglucosaminyl residues.

View complete list of fluorescent dye-conjugated lectins ›

Pierce™ Microplate BCA Protein Assay Kit - Reducing Agent Compatible Thermo Scientific™

This Pierce Microplate BCA Protein Assay Kit is the reducing agent-compatible version of our popular Pierce BCA Protein Assay. The kit enables you to measure protein concentration in samples that contain disulfide reducing agents and comes with twenty 96-well microplates.

Compare all available BCA protein assays ›

Features of the Microplate BCA Protein Assay Kit—Reducing Agent Compatible include:
Compatibility—assay samples that contain up to 5 mM DTT, 35 mM BME, or 10 mM TCEP
BCA technology—only a slight modification of the standard BCA Protein Assay protocol (15-min incubation with Compatibility Reagent); no precipitation steps required
Sample volume—requires only 25 µL (standard kit) or less than 10 µL (microplate kit) of sample
Colorimetric—measure with a standard spectrophotometer or plate reader (562 nm)
Uniformity—exhibits less protein-to-protein variation than dye-binding methods (Bradford)
Assay range—linear working range for BSA of 125 to 2000 µg/mL

The BCA Protein Assay Kit—Reducing Agent Compatible (BCA-RAC) provides all of the advantages of the original BCA assay, plus compatibility with disulfide reducing agents at concentrations routinely used in protein sample buffers. This special adaptation of the popular Pierce BCA Protein Assay method enables accurate protein concentration measurement for samples containing DTT, BME, or TCEP. This reducing agent-compatible BCA kit extends the already broad reagent compatibility of the BCA Protein Assay protocol to include nearly all types of components commonly present in protein research samples.

Related products
Pierce BCA Protein Assay Kit - Reducing Agent Compatible
96-Well Plates for Pierce BCA-RAC Assay
Ampule Breakers

Pierce™ BCA Protein Assay Kit Thermo Scientific™

Pierce BCA Protein Assay Kit is a two-component, high-precision, detergent-compatible protein assay for determination of protein concentration. Pierce BCA reagents provide accurate determination of protein concentration with most sample types encountered in protein research. The Pierce BCA assay can be used to assess yields in whole cell lysates, affinity-column fractions, purified proteins samples, as well as to monitor protein contamination in industrial applications. Compared to most dye-binding methods, the BCA assay is affected much less by protein compositional differences, providing greater concentration accuracy.

Compare all available BCA protein assays ›

Features of the BCA Protein Assay Kit include:
Colorimetric—estimate visually or measure with a standard spectrophotometer or plate reader at 562 nm
Uniformity—exhibits less protein-to-protein variation than dye-binding methods (Bradford)
Compatibility—unaffected by typical concentrations of most ionic and non-ionic detergents
Assay time—30-min incubation; much easier and four times faster than classical Lowry methods
Assay range—linear working range for BSA of 20 to 2000 µg/mL
Sensitivity—detect down to 5 µg/mL with enhanced protocol

How the assay works
The BCA Protein Assay combines the well-known reduction of Cu2+ to Cu1+ by protein in an alkaline medium with the highly sensitive and selective colorimetric detection of the cuprous cation (Cu1+) by bicinchoninic acid (BCA). The first step is the chelation of copper with protein in an alkaline environment to form a light blue complex. In this reaction, known as the biuret reaction, peptides containing three or more amino acid residues form a colored chelate complex with cupric ions in an alkaline environment containing sodium potassium tartrate.

In the second step of the color development reaction, BCA reacts with the reduced (cuprous) cation that was formed in step one. The intense purple-colored reaction product results from the chelation of two molecules of BCA with one cuprous ion. The BCA/copper complex is water soluble and exhibits a strong linear absorbance at 562 nm with increasing protein concentrations. The complex is approximately 100 times more sensitive (lower limit of detection) than the pale blue color of the first reaction.

The reaction is strongly influenced by four amino acid residues (cysteine, cystine, tyrosine, and tryptophan) in the amino acid sequence of the protein. However, unlike Coomassie dye-binding methods, the universal peptide backbone also contributes to color formation, helping to minimize variability caused by protein compositional differences.
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