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HCS LipidTOX™ Phospholipidosis and Steatosis Detection Kit, for high-content screening, for cellular imaging (Invitrogen™)

HCS LipidTOX™ Phospholipidosis and Steatosis Detection Kit offers a complete set of reagents for performing image-based high-content screening (HCS) assays to characterize phospolipidosis and steatosis, toxic side effects on lipid metabolism that can be triggered by drugs or other compounds. The kit supplies LipidTOX™ Red phospholipid stain and LipidTOX™ Green neutral lipid stain, which can be used sequentially for the analysis of phospholipidosis and steatosis, respectively, or can be used in isolation for single-parameter analysis. Both fluorescent probes can be easily detected with fluorescence microscopes and can be quantified with stand-alone image analysis software or the built-in image analysis software of most HCS readers. In addition to the phospholipid and neutral lipid stains, this kit supplies the nuclear stain Hoechst 33342, propranolol and cyclosporin A (control compounds), and DMSO. Sufficient reagents are supplied for 2, 96-well plates (H34157) or 10, 96-well plates (H34158).

eBioscience™ Human Regulatory T Cell Staining Kit #2 (Invitrogen™)

This Human Regulatory T cell Staining Kit contains all of the buffers and monoclonal antibodies for CD4, CD25, and Foxp3 necessary to successfully stain and identify regulatory T cells from human peripheral blood cells.

The RPA-T4 monoclonal antibody reacts with human CD4, a 59 kDa glycoprotein found on the surface of the majority of thymocytes, a subset of mature T cells (T helper cells), and at lower levels on monocytes. The BC96 monoclonal antibody reacts with human CD25 (also known as interleukin-2 receptor alpha, IL-2R alpha), a 55 kDa surface protein expressed by early progenitors of T cells and B cells, by mature, activated T cells and B cells, and at constitutively high levels on regulatory T cells. The PCH101 monoclonal antibody reacts with the amino terminus of human Foxp3 also known as FORKHEAD BOX P3, SCURFIN, and JM2. Foxp3 is a 49-55 kDa protein and a member of the forkhead/winged-helix family of transcription factors. It was identified as the gene responsible for the X-linked lymphoproliferative disease observed in scurfy (sf) mice and in the human disorder, X-linked autoimmunity-allergic dysregulation syndrome (XLAAD). Constitutive expression of Foxp3 mRNA has been shown in CD4+CD25+ regulatory T cells (Treg), and ectopic expression of Foxp3 in CD4+CD25- cells imparts a Treg phenotype in these cells.

Reactivity/Species
Human

Reported Application
Intracellular Staining Followed by Flow Cytometric Analysis

Staphylococcus aureus (Wood strain without protein A) BioParticles™, Alexa Fluor™ 488 conjugate (Invitrogen™)

The Molecular Probes® BioParticles® product line consists of a series of fluorescently labeled, heat- or chemically killed bacteria and yeast in a variety of sizes, shapes, and natural antigenicities. These fluorescent BioParticles® products have been employed to study phagocytosis by fluorescence microscopy, quantitative spectrofluorometry, and flow cytometry.

We offer E. coli (K-12 strain), S. aureus (Wood strain, without protein A) and zymosan (S. cerevisiae) BioParticles® conjugates covalently labeled with a variety of different fluorophores (special care has been taken to remove free dye after conjugation). Unlike the fluorescence of fluorescein-labeled BioParticles® conjugates, which is partially quenched in acidic environments, the fluorescence of the Alexa Fluor®, BODIPY® FL, tetramethylrhodamine and Texas Red® dye conjugates is uniformly intense over the pH range from 3 to 10.

BioParticles Specifications:
• Label (Ex/Em): Alexa Fluor® 488 (~495/519 nm)
• Particle: S. aureus (Wood strain, without protein A)
Opsonizing reagent available


Using BioParticles Products
BioParticles® conjugates are provided as lyophilized powders. There are approximately 3 x 108 E. coli or S. aureus particles per mg solid and approximately 2 x 107 zymosan particles per mg solid. BioParticles® conjugates can be reconstituted in the buffer of your choice for use in phagocytosis assays. The fluorescence of BioParticles® conjugates that are bound to the surface of the cell (but not internalized) can be quenched by ethidium bromide, trypan blue, or other quenchers. In addition to cellular applications, fluorescent BioParticles® conjugates may be effective as flow cytometry calibration references when sorting bacteria and yeast mutants. These small particles may also be useful references for light scattering studies because their sizes and shapes differ in characteristic ways.

Find More BioParticles® Products
We offer a large range of dye-labeled and unlabeled E. coli (K-12 strain), S. aureus (Wood strain, without protein A), and zymosan (S. cerevisiae) BioParticles® products. Find out about these products and their applications by reviewing Probes for Following Receptor Binding and Phagocytosis—Section 16.1 in the Molecular Probes® Handbook.

For pH-sensitive endocytosis assays, see our pHrodo™ BioParticles® conjugates.

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

eBioscience™ Human/Non-Human Primate Regulatory T Cell Staining Kit #1 (Invitrogen™)

This Human/Non-human Primate Regulatory T cell Staining Kit contains all of the buffers and monoclonal antibodies for CD4, CD25, and Foxp3 necessary to successfully stain and identify regulatory T cells from human, rhesus, cynomolgus, or chimpanzee peripheral blood cells. This kit may also be used to identify regulatory T cells in baboons, using the CD25 and Foxp3 monoclonal antibodies, however, the monoclonal antibody for CD4 that is included in this kit does not crossreact with baboon.

The OKT4 monoclonal antibody reacts with human, rhesus, cynomolgus, and chimpanzee CD4, a 59 kDa glycoprotein found on the surface of the majority of thymocytes, a subset of mature T cells (T helper cells), and at lower levels on monocytes. The BC96 monoclonal antibody reacts with human, rhesus, cynomolgus, chimpanzee, and baboon CD25 (also known as interleukin-2 receptor alpha, IL-2R alpha), a 55 kDa surface protein expressed by early progenitors of T cells and B cells, by mature, activated T cells and B cells, and at constitutively high levels on regulatory T cells. The PCH101 monoclonal antibody reacts with the amino terminus of human, rhesus, cynomolgus, chimpanzee, and baboon Foxp3 also known as FORKHEAD BOX P3, SCURFIN, and JM2. Foxp3 is a 49-55 kDa protein and a member of the forkhead/winged-helix family of transcription factors. It was identified as the gene responsible for the X-linked lymphoproliferative disease observed in scurfy (sf) mice and in the human disorder, X-linked autoimmunity-allergic dysregulation syndrome (XLAAD). Constitutive expression of Foxp3 mRNA has been shown in CD4+CD25+ regulatory T cells (Treg), and ectopic expression of Foxp3 in CD4+CD25- cells imparts a Treg phenotype in these cells.

Reactivity/Species
Human, Monkey

Reported Application
Intracellular Staining Followed by Flow Cytometric Analysis

HCS LipidTOX™ Green Phospholipidosis Detection Reagent (1000X), for cellular imaging (Invitrogen™)

The HCS LipidTOX™ Green phospholipidosis detection reagent was developed to characterize the potentially toxic side effects of compounds on lipid metabolism in mammalian cell lines. This reagent is easily detected by fluorescence microscopy or an HCS reader equipped with filters that can detect excitation/emission maxima ~495/525 nm. LipidTOX™ phospholipidosis detection reagents do not affect the normal growth of cells, and the staining observed after incubation in live cells is maintained after formaldehyde fixation. This probe is compatible with the HCS LipidTOX™ neutral lipid stains (H34475, H34476, and H34477).

eBioscience™ Human Regulatory T Cell Staining Kit (Invitrogen™)

This Human Regulatory T cell Staining Kit contains all of the buffers and monoclonal antibodies for CD4, CD25, and Foxp3 necessary to successfully stain and identify regulatory T cells from human peripheral blood cells.

The RPA-T4 monoclonal antibody reacts with human CD4, a 59 kDa glycoprotein found on the surface of the majority of thymocytes, a subset of mature T cells (T helper cells), and at lower levels on monocytes. The BC96 monoclonal antibody reacts with human CD25 (also known as interleukin-2 receptor alpha, IL-2R alpha), a 55 kDa surface protein expressed by early progenitors of T cells and B cells, by mature, activated T cells and B cells, and at constitutively high levels on regulatory T cells. The PCH101 monoclonal antibody reacts with the amino terminus of human Foxp3 also known as FORKHEAD BOX P3, SCURFIN, and JM2. Foxp3 is a 49-55 kDa protein and a member of the forkhead/winged-helix family of transcription factors. It was identified as the gene responsible for the X-linked lymphoproliferative disease observed in scurfy (sf) mice and in the human disorder, X-linked autoimmunity-allergic dysregulation syndrome (XLAAD). Constitutive expression of Foxp3 mRNA has been shown in CD4+CD25+ regulatory T cells (Treg), and ectopic expression of Foxp3 in CD4+CD25- cells imparts a Treg phenotype in these cells.

Reactivity/Species
Human

Reported Application
Intracellular Staining Followed by Flow Cytometric Analysis

Staphylococcus aureus (Wood strain without protein A) BioParticles™, Alexa Fluor™ 594 conjugate (Invitrogen™)

The Molecular Probes® BioParticles® product line consists of a series of fluorescently labeled, heat- or chemically killed bacteria and yeast in a variety of sizes, shapes, and natural antigenicities. These fluorescent BioParticles® products have been employed to study phagocytosis by fluorescence microscopy, quantitative spectrofluorometry, and flow cytometry.

We offer E. coli (K-12 strain), S. aureus (Wood strain, without protein A) and zymosan (S. cerevisiae) BioParticles® conjugates covalently labeled with a variety of different fluorophores (special care has been taken to remove free dye after conjugation). Unlike the fluorescence of fluorescein-labeled BioParticles® conjugates, which is partially quenched in acidic environments, the fluorescence of the Alexa Fluor®, BODIPY® FL, tetramethylrhodamine and Texas Red® dye conjugates is uniformly intense over the pH range from 3 to 10.

BioParticles Specifications:
• Label (Ex/Em): Alexa Fluor® 594 (~590/617 nm)
• Particle: S. aureus (Wood strain, without protein A)
Opsonizing reagent available


Using BioParticles Products
BioParticles® conjugates are provided as lyophilized powders. There are approximately 3 x 108 E. coli or S. aureus particles per mg solid and approximately 2 x 107 zymosan particles per mg solid. BioParticles® conjugates can be reconstituted in the buffer of your choice for use in phagocytosis assays. The fluorescence of BioParticles® conjugates that are bound to the surface of the cell (but not internalized) can be quenched by ethidium bromide, trypan blue, or other quenchers. In addition to cellular applications, fluorescent BioParticles® conjugates may be effective as flow cytometry calibration references when sorting bacteria and yeast mutants. These small particles may also be useful references for light scattering studies because their sizes and shapes differ in characteristic ways.

Find More BioParticles® Products
We offer a large range of dye-labeled and unlabeled E. coli (K-12 strain), S. aureus (Wood strain, without protein A), and zymosan (S. cerevisiae) BioParticles® products. Find out about these products and their applications by reviewing Probes for Following Receptor Binding and Phagocytosis—Section 16.1 in the Molecular Probes® Handbook.

For pH-sensitive endocytosis assays, see our pHrodo™ BioParticles® conjugates.

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

HCS LipidTOX™ Red Phospholipidosis Detection Reagent (1000X), for cellular imaging (Invitrogen™)

The HCS LipidTOX™ Red phospholipidosis detection reagent was developed to characterize the potentially toxic side effects of compounds on lipid metabolism in mammalian cell lines. This reagent is easily detected by fluorescence microscopy or an HCS reader equipped with filters that can detect excitation/emission maxima ~595/615 nm. LipidTOX™ phospholipidosis detection reagents do not affect the normal growth of cells, and the staining observed after incubation in live cells is maintained after formaldehyde fixation. This probe is compatible with the HCS LipidTOX™ neutral lipid stains (H34475, H34476, and H34477).

XTT (2,3-Bis-(2-Methoxy-4-Nitro-5-Sulfophenyl)-2H-Tetrazolium-5-Carboxanilide) (Invitrogen™)

XTT is used to assess cell viability as a function of redox potential. Actively respiring cells convert the water-soluble XTT to a water-soluble, orange colored formazan product. Unlike MTT, XTT does not require solubilization prior to quantitation, thereby reducing the assay time in many viability assay protocols. Moreover, the sensitivity of the XTT reduction assay is reported to be similar to or better than that of the MTT reduction assay.

SuperPicture™ Polymer Detection Kit, HRP, broad spectrum

The SuperPicture™ Polymer Detection kit features our proprietary and enhanced HRP polymer technology to provide excellent sensitivity, high specificity, and a shorter
protocol than conventional immunohistochemical staining methods.

Our new polymer conjugation methods have resulted in a versatile polymer capable of intense nuclear, cytoplasmic, and membrane antigen staining. The polymer conjugate contains multiple HRP molecules to provide excellent sensitivity. The polymer staining method is faster and easier than a conventional LAB-SA protocol because it uses an HRP polymer conjugated to the second antibody and eliminates one incubation step. In addition, SuperPicture™ Polymer kits do not contain biotin nor streptavidin, so potential background due to endogenous biotin activity is completely avoided. Another advantage of SuperPicture™ Polymer detection system is the shorter polymer incubation time; the this system only requires 10 minutes, while leading competitors’ polymers require 30 minutes.

Reactivity: Mouse, Rabbit, Rat, G. Pig

Applications: Immunohistochemistry (FFPE)

Histostain-Plus IHC Kit, HRP, broad spectrum

Full Product Description
Histostain-Plus kits use 2nd generation LAB-SA technology and are ideal for general immunohistochemical applications. Histostain-Plus kits offer superior sensitivity without compromising background. They are designed to detect mouse, rabbit, rat, or guinea pig primary antibodies that are bound to human tissue or cell antigens. Histostain-Plus kits are excellent for frozen and formalin-fixed, paraffin-embedded tissue sections, cell smears, etc. These kits are very easy to use: all components are liquid (except Fast-Red) and come in dropper bottles. No reconstitution, no pipetting, no measuring is necessary, and the enzyme is conjugated to streptavidin to eliminate the preincubation complexing step. DAB chromogen yields a brown stain, AEC yields a deep red stain, and Fast-Red yields a bright red stain.

Reactivity: Mouse, Rabbit, Rat, Guinea Pig

Applications: Immunohistochemistry (IHC)

Pacific Blue™ Annexin V/SYTOX™ AADvanced™ Apoptosis Kit, for flow cytometry (Invitrogen™)

This product detects the externalization of phosphatidylserine in apoptotic cells using recombinant annexin V conjugated to violet-fluorescent Pacific Blue™ dye and dead cells using SYTOX™ AADvanced™ stain. After staining a cell population with Pacific Blue™, annexin V and SYTOX™ AADvanced™, apoptotic cells show violet fluorescence, dead cells show red fluorescence, and live cells show little or no fluorescence. These populations are easily distinguished by a flow cytometer with the 405 nm and 488 nm lines for excitation. There is very little spectral overlap between the two dyes, therefore very little compensation is needed. Each kit contains sufficient reagents for approximately 50 flow cytometry tests.

View a selection guide for all apoptosis assays for flow cytometry.

pHrodo™ Red Phagocytosis Particle Labeling Kit for Flow Cytometry (Invitrogen™)

• Specifically detect phagocytosis and endocytosis with pH-sensitive fluorogenic dye - discriminate endcytosed from adherent and extracellular particles
• Reduced signal variability and improved timing in sensitive experiments - no need for wash steps or quencher dye
• Bright red fluorescence - conveniently multiplex with green dyes such as GFP, Fluo-4, or calcein

View a selection guide for all pHrodo Indicators used in imaging, flow cytometry and microplate assays.

The pHrodo™ Red dye gives faster and more accurate results than any other phagocytosis assay

The new Molecular Probes™ proprietary pH-sensitive rhodamine-based pHrodo™ Red dye is non-fluorescent at neutral pH, but turns bright red upon acidification. Because it is both fluorogenic and pH-sensitive, the pHrodo™ Red dye can be used as a specific sensor of phagocytic events; acidification of the phagosome following phagocytosis is marked by red fluorescence. It is therefore an ideal tool with which to study phagocytosis and its regulation by drugs and/or environmental factors.

Rapid and reproducible

Wash steps and quencher dyes are not needed since the pHrodo™ Red dye is non-fluorescent outside the cell, making the staining protocol simple and fast. The elimination of wash steps and quencher dyes also improves assay reproducibility, particularly in plate reader-based assays.

Flexible applications

Use ready-made pHrodo™ Red BioParticles™ conjugates for convenient analysis of phagocytosis of Gram-positive or Gram-negative bacteria, or the amine-reactive pHrodo™ Red SE form for labeling microorganisms or proteins of your choice.

Compatible with multiple platforms
pHrodo™ Red dye conjugates can be used in plate readers, fluorescence microscopy imaging, and flow cytometry applications. The pHrodo™ Red Phagocytosis Particle Labeling Kit and the pHrodo™ Red E. coli BioParticles™ Phagocytosis Kit for flow cytometry were designed for rapid and convenient measurements of phagocytic activity in whole blood samples by flow cytometry. The kits include all the reagents required for assessing particle ingestion and red blood cell lysis. The Labeling Kit also contains the reagents required for labeling microorganisms with the pHrodo™ Red dye. Sufficient reagents are provided in both kits for approximately 100 assays.

The optimal absorption and fluorescence emission maxima of the pHrodo™ Red BioParticles™ conjugate is approximately 560 nm and 585 nm, respectively. However, the fluorophore is readily excited with the 488 nm argon-ion laser installed on most flow cytometers.