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Anti-Human Foxp3 Staining Set APC, eBioscience™ Invitrogen™

This Anti-Human Foxp3 Staining Set contains the buffers and monoclonal antibody necessary to successfully stain and identify Foxp3 cells. The 236A/E7 monoclonal antibody reacts with 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.

The 236A/E7 antibody has been shown to crossreact with rhesus macaque, sooty mangabey, and cynomolgus Foxp3 236A/E7 recognizes a different epitope of Foxp3 than PCH101 and 150D/E4.

Isotype
IgG1

Laser
Red Laser

Emit
660 nm

Excite
633 - 647 nm

Reported Application
Intracellular Staining Followed by Flow Cytometric Analysis

Anti-Mouse/Rat Foxp3 Staining Set APC, eBioscience™ Invitrogen™

The FJK-16s antibody reacts with mouse, rat, dog, porcine, and bovine Foxp3 also known as FORKHEAD BOX P3, SCURFIN, and JM2; cross reactivity of this antibody to other proteins has not been determined. Foxp3 a 49-55 kDa protein, is a member of the forkhead/winged-helix family of transcriptional regulators, and was identified as the gene defective in ‘scurfy’ (sf) mice. Constitutive high expression of Foxp3 mRNA has been shown in CD4+CD25+ regulatory T cells (Treg cells), and ectopic expression of Foxp3 in CD4+CD25- cells imparts a Treg phenotype in these cells.

Immunoblotting with FJK-16s antibody has mapped the epitope to amino acids 75-125 of the mouse Foxp3 protein. In the human, this region has been shown to be alternatively spliced at the mRNA level. Both the alternatively-spliced and non-spliced isoforms are present in the CD4+CD25+ subset of lymphocytes. Preliminary RT-PCR experiments have not revealed this alternatively-spliced isoform in mouse splenocytes, suggesting different gene regulation in the mouse and human.

Intracellular staining of mouse splenocytes with FJK-16s using the PE anti-mouse/rat Foxp3 Staining Set and protocol reveals approximately 2% of total cells in the C57Bl/6 strain and approximately 3-5% in the BALB/c mouse strain. Multicolor flow cytometric analysis demonstrates approximately 90% of the CD4+CD25+ cells and 4% of the CD4+CD25- cells staining with FJK-16s. B220+, CD11b+, CD11c+, and Ly6G/Gr-1+ cells do not show significant co-staining with FJK-16s.

Please see our FAQ regarding the usage of eBioscience Foxp3 reagents.

Not included:
Fc Block (cat. 14-0161)
Flow Cytometry Staining Buffer (cat. 00-4222)
Rat IgG2a isotype Control (cat. 12-4321, 11-4321, or 17-4321)

Isotype
IgG2a, kappa

Laser
Red Laser

Emit
660 nm

Excite
633 - 647 nm

Reported Application
Intracellular Staining Followed by Flow Cytometric Analysis

Pacific Orange™ Antibody Labeling Kit Invitrogen™

Molecular Probes™ Antibody Labeling Kits provide a convenient means to label small amounts of antibodies with the violet light-excitable Pacific Orange™ or Pacific Blue™ dyes. This kit is optimized for labeling 100 µg of antibody per reaction. Comparably small amounts of other proteins (>40 kDa) can also be labeled.

View a selection guide for all Antibody Labeling Kits.

View the Fluorophore Selection Guide.

The kit contains everything you need to perform five separate labeling reactions as well as to purify the resulting conjugates. Conjugates are ideal for multiple applications, including flow cytometry, fluorescent microscopy, immunohistochemistry, primary detection, ELISAs, immunocytochemistry, indirect FISH, and more.

Important features of antibody labeling kits:
• Pacific Orange™ has an excitation and emission maximum of 400/551 nm
• Labeled proteins typically ready to use in 90 min (~15 min hands-on time)
• Useful for labeling 100 µg of protein
• Optimized for small-scale labeling of any protein >40 kDa
• Purified using convenient spin filters
• Stabilizing proteins must be removed from the sample before labeling
• Includes detailed instructions for determining degree of labeling (DOL)


Better results and workflows with primary labeled antibodies
A primary antibody directly labeled with a fluorophore often produces lower background fluorescence and less nonspecific binding. Further, multiple primary antibodies of the same isotype or derived from the same species can easily be used in the same experiment if they are directly labeled with compatible fluorophores.

Learn more about protein and antibody labeling
We offer a wide selection of Molecular Probes™ antibody and protein labeling kits to fit your starting material and your experimental setup. See Antibody Labeling from A to Z or use our Labeling Chemistry Selection Tool for other choices. To learn more about our various kits read Kits for Labeling Proteins and Nucleic Acids—Section 1.2 in the Molecular Probes™ Handbook.

We'll make a custom antibody conjugate for you
If you can't find what you're looking for in our stocked list, we'll prepare a custom antibody conjugate for you. Our custom conjugation service is efficient and confidential, and we stand by the quality of our work. We are ISO 9001:2000 certified.

Anti-Human Foxp3 Staining Set APC, eBioscience™ Invitrogen™

This Anti-Human Foxp3 Staining Set contains the buffers and monoclonal antibody necessary to successfully stain and identify Foxp3 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.

The PCH101 antibody crossreacts with rhesus, chimpanzee, and cynomolgus Foxp3 PCH101 recognizes a different epitope of Foxp3 than clones 236A/E7 and 150D/E4.

Isotype
IgG2a, kappa

Laser
Red Laser

Emit
660 nm

Excite
633 - 647 nm

Reported Application
Intracellular Staining Followed by Flow Cytometric Analysis

ELISA TMB Stabilized Chromogen Invitrogen™

ELISA TMB Stabilized Chromogen is a ready-to-use one-component horseradish peroxidase substrate (HRP) for ELISA plate applications. It can be used in both kinetic assays (optical density monitored over several time points) and in endpoint assays. The solution is colorless or pale blue prior to use. In the presence of peroxidase, the solution develops a blue reaction product. The optical density of the solution is measured at two wavelengths: 492 nm and 650 nm.

When color formation is monitored at an endpoint, an equal volume of an acidic stop solution should be added to the wells. Addition of a stop solution results in the rapid formation of a yellow end product. The optical density of this yellow end product is measured at 450 nm.

For a complete kit of ELISA buffers, see ELISA Buffer Kit.
View all available buffers and reagents for ELISA applications.

Oncomine™ Gene Browser, Academic License

Oncomine™ Gene Browser, Academic, is a web-based software tool for academic researchers that provides easy access to a comprehensive gene profile across thousands of cancer patient genomes. The rich dataset is presented in a visual and dynamic environment that easily conveys significant gene mutations, expression, and DNA copy number changes. It also enables customers to quickly and easily ask questions across hundreds of cancer subtypes for any gene. Designed for a broad audience, the Oncomine™ Gene Browser is fast, easy to use, and provides powerful insight into any gene target by deriving knowledge from the world’s oncology experiments.

• Profile your critical targets and biomarkers across tens of thousands of retrospective de-identified patient tumor genomes online, in minutes
• Rapidly prioritize targets based on mutations, copy number variants, and differential expression
• Identify optimal patient segments for your clinical development based on biomarker frequencies in well-characterized patient populations

Benefits
A key benefit of Oncomine™ Gene Browser is the fast and comprehensive evaluation of your oncology gene targets within the world's oncology experimental data. No longer relying on one or two research publications for validation or being restricted to author interpretation and focus, an unbiased approach to evaluating your gene targets can be performed in minutes, often yielding insights into new cancer subtypes that can expand your target clinical population.

Oncomine™ Gene Browser empowers users to assess mutations, gene expression, DNA copy number, cancer subtypes, normal tissue expression, and pick cell line models. In a nutshell, millions of dollars worth of experiments are at your fingertips and can be 'performed' in silico so that you can design and execute fewer, more focused wet lab experiments on the targets and indications that have a greater chance for success.

Learn more about Oncomine™ Gene Browser.

Oncomine™ Gene Browser, Academic, can be licensed for various user numbers, up to an institutional license with an unlimited number of registered users.

Note: This product is for academic and nonprofit cancer researchers only. If you are a commercial/for-profit cancer researcher, please contact one of our knowledgable sales specialists at compendiasales@lifetech.com.

How to Order
Order this product as you would any standard product. You will receive an email with your user name and temporary password, which you can then use to log in and get started.

Dynabeads™ EPEC/VTEC O26 Applied Biosystems™

For the rapid selective concentration of E.coli 026. Method can be performed manually or can be automated by using a Thermo Scientific KingFisher mL Food Protection Purification System.

This Product Contains: Dynabeads coated with anti-E.coli O26 antibodies

Sensitivity: Sensitivity is very high and enables detection of 1 viable organism in 25g of sample or as few as 100 organisms / ml after pre enrichment

Anti-Human Foxp3 Staining Set FITC, eBioscience™ Invitrogen™

This Anti-Human Foxp3 Staining Set contains the buffers and monoclonal antibody necessary to successfully stain and identify Foxp3 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.

The PCH101 antibody crossreacts with rhesus, chimpanzee, and cynomolgus Foxp3 PCH101 recognizes a different epitope of Foxp3 than clones 236A/E7 and 150D/E4.

Isotype
IgG2a, kappa

Laser
Blue Laser

Emit
518 nm

Excite
488 nm

Reported Application
Intracellular Staining Followed by Flow Cytometric Analysis

Click-iT™ Plus EdU Alexa Fluor™ 594 Flow Cytometry Assay Kit Invitrogen™

The Click-iT Plus EdU Alexa Fluor 594 Flow Cytometry Assay Kit provides a simplified, more robust assay for analyzing DNA replication in proliferating cells compared to traditional BrdU methods. Newly synthesized DNA is analyzed using the 532- or 561-nm laser of the flow cytometer. The Click-iT Plus formulation is compatible with standard fluorophores, including R-PE and R-PE tandems, as well as fluorescent proteins.

• Multiplexable—compatible with R-PE (and tandems) and fluorescent proteins
• Accurate—superior results compared to BrdU assays
• Fast—results in as little as 60 minutes

View selection guide for all Click-iT EdU and Click-iT Plus EdU assays for flow cytometry.

Multiplexable
Click-iT Plus Alexa Fluor 594 EdU assays can be used in conjunction with R-PE and R-PE tandems, as well as fluorescent proteins such as GFP and mCherry. The Alexa Fluor 594 labeling reagent is excited readily at either 532 or 561 nm and emits at 615 nm.

An Advanced Method Giving You Results Superior to BrdU
The most accurate method of proliferation analysis is direct measurement of DNA synthesis. Originally, this was performed through incorporation of radioactive nucleosides. This method was replaced by antibody-based detection of the nucleoside analog bromodeoxyuridine (BrdU). The Click-iT Plus EdU Flow Cytometry assay is a novel alternative to the BrdU assay. EdU (5-ethynyl-2´-deoxyuridine) is a thymidine analog that is incorporated into DNA during active DNA synthesis. Detection is based on click chemistry, which is a copper-catalyzed covalent reaction between an azide and an alkyne. In this application, the alkyne is found in the ethynyl moiety of EdU, while the azide is coupled to the Alexa Fluor dye. Standard flow cytometry methods are used for determining the percentage of S-phase cells in the population.

Mild Conditions Allow Use with Cell Cycle Dyes and Antibodies
The small size of the dye azide allows for efficient detection of the incorporated EdU using mild conditions, while standard aldehyde-based fixation and detergent permeabilization are sufficient for the Click-iT Plus detection reagent to gain access to the DNA. This is in contrast to BrdU assays that require DNA denaturation (using HCl, heat, or digestion with DNase) to expose the BrdU so that it can be detected with an anti-BrdU antibody. Sample processing for the BrdU assay can result in signal alteration of the cell cycle distribution, as well as destruction of antigen recognition sites when using the HCl method. In contrast, the easy-to-use Click-iT Plus EdU assay is compatible with cell cycle dyes. The Click-iT Plus EdU assay can also be multiplexed with antibodies against surface and intracellular markers, as well as conjugates labeled with standard fluorophores including R-PE, R-PE tandems, and fluorescent proteins (GFP and mCherry).

Quick and Simple Protocol
The Click-iT Plus EdU protocol is based on the aldehyde fixation and detergent permeabilization steps for immunohistochemical antibody labeling. However, EdU is compatible with other fixation/permeabilization agents including saponin and methanol. In just five steps you’ll be ready to analyze your cell proliferation data:

1. Treat cells with EdU.
2. Fix and permeabilize cells.
3. Detect S-phase cells with Click-iT Plus detection cocktail for 30 min.
4. Wash once.
5. Analyze.

Results can be seen in as little as 60 minutes in some circumstances, but we recommend 90 minutes for all applications.

Anti-Mouse/Rat Foxp3 Staining Set PE, eBioscience™ Invitrogen™

The FJK-16s antibody reacts with mouse, rat, dog, porcine, and bovine Foxp3 also known as FORKHEAD BOX P3, SCURFIN, and JM2; cross reactivity of this antibody to other proteins has not been determined. Foxp3 a 49-55 kDa protein, is a member of the forkhead/winged-helix family of transcriptional regulators, and was identified as the gene defective in ‘scurfy’ (sf) mice. Constitutive high expression of Foxp3 mRNA has been shown in CD4+CD25+ regulatory T cells (Treg cells), and ectopic expression of Foxp3 in CD4+CD25- cells imparts a Treg phenotype in these cells.

Immunoblotting with FJK-16s antibody has mapped the epitope to amino acids 75-125 of the mouse Foxp3 protein. In the human, this region has been shown to be alternatively spliced at the mRNA level. Both the alternatively-spliced and non-spliced isoforms are present in the CD4+CD25+ subset of lymphocytes. Preliminary RT-PCR experiments have not revealed this alternatively-spliced isoform in mouse splenocytes, suggesting different gene regulation in the mouse and human.

Intracellular staining of mouse splenocytes with FJK-16s using the PE anti-mouse/rat Foxp3 Staining Set and protocol reveals approximately 2% of total cells in the C57Bl/6 strain and approximately 3-5% in the BALB/c mouse strain. Multicolor flow cytometric analysis demonstrates approximately 90% of the CD4+CD25+ cells and 4% of the CD4+CD25- cells staining with FJK-16s. B220+, CD11b+, CD11c+, and Ly6G/Gr-1+ cells do not show significant co-staining with FJK-16s.

Please see our FAQ regarding the usage of eBioscience Foxp3 reagents.

Not included:
Fc Block (cat. 14-0161)
Flow Cytometry Staining Buffer (cat. 00-4222)
Rat IgG2a isotype Control (cat. 12-4321, 11-4321, or 17-4321)

Isotype
IgG2a, kappa

Laser
Blue Laser

Emit
575 nm

Excite
488 nm

Reported Application
Intracellular Staining Followed by Flow Cytometric Analysis

Custom OpenArray™ Preamp Pool Applied Biosystems™

Custom OpenArray® Preamp Pool

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

Oncomine™ Research Premium Edition License

Oncomine™ Research Premium Edition is a subscription-based software tool for academic researchers that provides additional advanced features and analyses over Oncomine™ Research Edition (a free edition available at www.oncomine.org). Oncomine™ Research Edition is a powerful web application that integrates and unifies high-throughput cancer profiling data so that target expression across a large number of cancer types and experiments can be accessed online, in seconds. Oncomine™ Research Edition includes basic analysis types such as cancer vs. normal, multi-cancer, and co-expression. It features gene and concept summaries, outlier analysis, meta-analysis, and meta-cancer outlier profile analysis (COPA). In addition, Oncomine™ Research Edition has an Oncomine™ support community site where users can share their experiences using this software tool.

Oncomine™ Research Premium Edition is a subscription version that allows users with advanced research needs to take advantage of additional features and analyses. These features include and both basic and advanced analyses such as cancer subtype, clinical outcome, patient treatment response, perturbation analyses, and drug sensitivity analyses. These analyses provide valuable insight into biology, regulation, pathways, drug response, and patient populations. In addition, the Premium Edition allows users to search for multiple genes at a time, export to Excel, PowerPoint, and SVG to share results, upload custom concepts, and access enhanced support, as well as search the Oncomine™ database for enrichment of user-defined gene signatures.

Oncomine™ Research Premium Edition features:

• Advanced search features, including multi-gene searching, allow you to visualize your data faster and easier
• My Concepts is a custom concept upload that makes your workflows more efficient by allowing you to quickly run an analysis on your specific signature
• Export results to Excel, PowerPoint, and SVG to perform additional analyses and easily share your findings; export nodes and edges files for concept association visualizations
• Sophisticated built-in analyses allow you to answer critical research questions about your data

Learn more about Oncomine™ Research Premium Edition.

Oncomine™ Research Premium Edition can be licensed for various user numbers, up to an institutional license with unlimited number of registered users. You must be a registered user of Oncomine™ Research Edition to upgrade.

Note: This product is for academic and nonprofit cancer researchers only. If you are a commercial/for-profit cancer researcher, please contact one of our knowledgable sales specialists at compendiasales@lifetech.com.

How to Order
If you are not already a registered user of the free version of the software, please visit www.oncomine.org to register for Oncomine™ Research Edition. You will receive an email with your user name and temporary password, which you can then use to log in a get started using the free edition. Once this is done, or if you are already a registered use of the free edition, simply order this product as you would any standard product. During checkout you will be asked to provide information about each user of the software. Within 1 business day your account will be upgraded to the Premium edition and you will see the added features of the Premium edition the next time you log in.

CaptureSelect™ IgG-CH1 Affinity Matrix Thermo Scientific™

CaptureSelect™ IgG-CH1 Affinity Matrix has been specifically designed for the purification of recombinant human Fab fragments and IgGs, recognizing all IgG subclasses (1, 2, 3 and 4) independent of the light chain subclass (Kappa/Lambda).

Features of this affinity matrix include:

• Platform approach for purification of IgGs and their Fab fragments
• No co-purification of overexpressed free lights chains due to heavy chain binding
• Excellent scalability
• Non-animal-derived

CaptureSelect™ IgG-CH1 Affinity Matrix purifies IgG, Fab, and Fab2 fragments directly from complex source materials in a single step with high purity and yield.

Free of Animal Components
Our CaptureSelect™ products are affinity ligands created by a proprietary technology based on Camelid-derived single domain antibody fragments. The ligand is a 14 kDa single domain fragment comprising the 3 CDRs that form the antigen binding domain, efficiently produced by the yeast Saccharomyces cerevisiae in a production process free of animal components.

Main Characteristics:

Matrix: agarose-based, aldehyde-activated
Average particle size: 65 ± 10 µm
Ligand: CaptureSelect™ IgG-CH1 affinity ligand
Ligand coupling method: aldehyde coupling via NH2 residues of the ligand
Binding capacity: 25–30 g IgG/L resin depending on flow rate, column height and contact time
Elution conditions: 20 mM citric acid, 150 mM NaCl, pH 3.5
Flow characteristics: 200–400 cm/h
Formulation buffer: 20%(v/v) ethanol

Dynabeads™ EPEC/VTEC O111 Applied Biosystems™

For the rapid selective concentration of E.coli 0111. Method can be performed manually or can be automated by using a Thermo Scientific KingFisher mL Food Protection Purification System.

This Product Contains: Dynabeads coated with anti-E.coli 0111 antibodies

Sensitivity: Sensitivity is very high and enables detection of 1 viable organism in 25g of sample or as few as 100 organisms / ml after pre enrichment
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