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Pierce™ Alkaline Phosphatase, Biotinylated (Thermo Scientific™)

Thermo Scientific Pierce Biotinylated Proteins include biotin-labeled proteins (BSA), enzymes (HRP, AP) and fluorophores (FITC) for use as controls or signal amplification in IHC via avidin-biotin complex (ABC) techniques.

Biotinylated AP is most commonly used in immunohistochemistry (IHC) to amplify the signal of biotinylated primary antibodies using the ABC staining method.

Biotinylated enzymes—biotin-labeled horseradish peroxidase (B-HRP), alkaline phosphatase (B-AP) and beta-galactosidase (B-bGal) for use in avidin-biotin complex (ABC) staining

Biotin, also known as vitamin H, is a small molecule (MW 244.3) that is present in tiny amounts in all living cells and is critical for a number of biological processes. The valeric acid side chain of the biotin molecule can be derivatized in order to incorporate various reactive groups that are used to attach biotin to other molecules. In the context of immunohistochemistry (IHC), biotin is conjugated to antibodies or to the enzyme reporters used to detect target antigens.

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5-CFDA, AM (5-Carboxyfluorescein Diacetate, Acetoxymethyl Ester) (Invitrogen™)

5-CFDA, AM is a cell-permeant esterase substrate that can serve as a viability probe that measures both enzymatic activity, which is require to activate its fluorescence, and cell-membrane integrity, which is required for intracellular retention of their fluorescent product. The electronically neutral AM ester of CFDA can be loaded into cells at lower concentrations than CFDA. Upon hydrolysis by intracellular esterases, this AM ester also yields carboxyfluorescein. As compared with fluorescein, carboxyfluorescein contains extra negative charges and is therefore better retained in cells.

Annexin V APC Ready Flow Conjugate (Invitrogen™)

Annexin V APC Ready Flow Conjugate is a bright, easy-to-use stain with high affinity for phosphatidylserine (PS), which becomes exposed on the outer leaflet of cells undergoing apoptosis. Because of this affinity, fluorescently labeled annexin V reagents are commonly used in apoptosis research. The Annexin V APC conjugate is excitable at 650 nm with an emission peak at 660 nm.

• Stable at room temperature
• Convenient, ready-to-use format—no need to dilute, weigh, or pipette
• Can be added directly to Ca2+-containing media or annexin binding buffer

Annexin V APC Ready Flow Conjugate is a ready-to-use, direct-to-sample apoptotic cell stain. Add one drop from the dropper bottle directly into 1 x 106 cells in 5 mM Ca2+-containing cell culture media or annexin binding buffer (Cat. Nos. V13246 or BMS500BB), incubate, and analyze. The traditional annexin staining protocol can be used as an alternative method by replacing the addition of 5 µL annexin conjugate with the addition of 1 drop from the Ready Flow dropper bottle.

CaptureSelect™ Biotin Anti-IgG-Fc (Multi-species) Conjugate (Thermo Scientific™)

CaptureSelect™ Biotin Anti-IgG-Fc (ms) Conjugate consists of a 13 kDa llama antibody fragment (affinity ligand) that specifically binds to the Fc part of IgG from multiple species. The affinity ligand is chemically conjugated to biotin via an appropriate spacer that retains the binding reactivity of the ligand when used in combination with streptavidin-based conjugates or streptavidin pre-coated surfaces.

The CaptureSelect™ Biotin Anti-IgG-Fc (ms) Conjugate allows you to:

Detect, quantitate, and characterize IgG antibodies from multiple species (all subclasses, IgG-specific), multi-species IgG-derived Fc fusion proteins, recombinant IgG antibodies, and total IgG in plasma and/or serum samples
Avoid cross-binding with other isotypes, such as IgA and IgM

Applications for CaptureSelect™ Biotin Anti-IgG-Fc (ms) Conjugate include ELISA, Western blot, Gyrolab™-based immunoassays, and label-free detection platforms such as those based on surface plasmon resonance (Biacore™ and IBIS-MX96 systems) and bio-layer interferometry (ForteBio™ Octet™ systems).

CaptureSelect™ Biotin Anti-Free LC-kappa (Human) Conjugate (Thermo Scientific™)

CaptureSelect™ Biotin Anti-Free LC-kappa (Hu) Conjugate consists of a 13 kDa llama antibody fragment (affinity ligand) that binds highly selectively to free human kappa light chains and dimers thereof. The functionality is based on recognition of an epitope on the human CL-kappa domain that is masked in intact antibodies and Fab fragments. The affinity ligand is chemically conjugated to biotin via an appropriate spacer that retains the binding reactivity of the ligand when used in combination with streptavidin-based conjugates or streptavidin pre-coated surfaces.

The CaptureSelect™ Biotin Anti-Free LC-kappa (Hu) Conjugate allows you to:

Detect, quantitate, and characterize free human kappa light chains in (crude) antibody preps
Avoid cross-binding with human kappa light chains present in intact and properly folded antibodies and Fab fragments of any human antibody isotype such as IgG, IgA, IgM, IgD, and IgE

Applications for CaptureSelect™ Biotin Anti-Free LC-kappa (Hu) Conjugate include ELISA, Western blot, Gyrolab™-based immunoassays, and label-free detection platforms such as those based on surface plasmon resonance (Biacore™ and IBIS-MX96 systems) and bio-layer interferometry (ForteBio™ Octet™ systems).

Fl - SRC 2-3 peptide, 100µM

LanthaScreen® fluorescent nuclear receptor coregulator peptides contain known interaction motifs and are labeled with fluorescein. These peptides are matched and validated to complement the LanthaScreen® TR-FRET Nuclear Receptor Coregulator Assays. Assays developed using these reagents enable primary or secondary screening of nuclear receptor agonists and/or antagonists.

How they work
Ligand binding to nuclear receptors causes conformational changes in the receptor, resulting in a cascade of events, including dissociation of repressor proteins, association of coactivator proteins, and assembly of pol II and other transcriptional factors for activation of target genes. TR-FRET based assays can be developed using the LanthaScreen® panel of fluorescein-labeled coregulator peptides to investigate conformational changes of nuclear receptors upon ligand binding, either by determining the affinity of ligand-bound receptor for different coregulator peptides, or by identifying additional agonists or antagonists via displacement or recruitment of a specific coregulator peptide. In the LanthaScreen® TR-FRET RXR beta Coactivator Assay, a terbium (Tb)-labeled anti-GST antibody is used to indirectly label GSTtagged RXR beta protein. An agonist (9-cis Retinoic Acid) added to the receptor which, upon ligand binding, causes a conformational change resulting in recruitment of the Fl-D22 coactivator peptide (Figure 1). The binding of the fluorescent coregulator peptide to RXR beta causes an increase in the TR-FRET emission ratio (Figure 2).

Contents and Storage:

LanthaScreen® fluorescent nuclear receptor coregulator peptides are supplied as 100 µM solutions in peptide-appropriate buffers. Store peptides at -20°C.

Annexin V Pacific Blue™ Ready Flow Conjugate (Invitrogen™)

Annexin V Pacific Blue Ready Flow Conjugate is a bright, easy-to-use stain with high affinity for phosphatidylserine (PS), which becomes exposed on the outer leaflet of cells undergoing apoptosis. Because of this affinity, fluorescently labeled annexin V reagents are commonly used in apoptosis research. The Annexin V Pacific Blue conjugate is excitable at 410 nm with an emission peak at 455 nm.

• Stable at room temperature
• Convenient, ready-to-use format—no need to dilute, weigh, or pipette
• Can be added directly to Ca2+-containing media or annexin binding buffer

Annexin V Pacific Blue Ready Flow Conjugate is a ready-to-use, direct-to-sample apoptotic cell stain. Add one drop from the dropper bottle directly into 1 x 106 cells in 5 mM Ca2+-containing cell culture media or annexin binding buffer (Cat. Nos. V13246 or BMS500BB), incubate, and analyze. The traditional annexin staining protocol can be used as an alternative method by replacing the addition of 5 µL annexin conjugate with the addition of 1 drop from the Ready Flow dropper bottle.

D-Myo-Inositol 1,4,5-Triphosphate, P4(5)-(1-(2-Nitrophenyl)ethyl) Ester, tris(triethylammonium) Salt (NPE-Caged Ins 1,4,5-P3) (Invitrogen™)

NPE-caged inositol 1,4,5-triphosphate is biologically inactive before photolysis. Flash photolysis of the caging group with UV illumination at 355 nm results in a rapid and highly localized release of free INS 1,4,5-P3 at the site of illumination.

CaptureSelect™ Human Fab-kappa Kinetics Biotin Conjugate (Thermo Scientific™)

CaptureSelect™ Human Fab-kappa Kinetics Biotin Conjugate is a bi-functional ligand construct consisting of 2 different 13 kDa llama antibody fragments (affinity ligands). The CaptureSelect™ anti-LC-kappa (Hu) ligand that specifically binds to the constant domain of human kappa light chains is genetically fused to the CaptureSelect™ anti-IgG-CH1 ligand that specifically binds to the CH1 domain of all four human IgG subclasses. The bi-functional affinity ligand is chemically conjugated to biotin via an appropriate spacer that retains the binding reactivity of the ligand when used in combination with streptavidin-based conjugates or streptavidin pre-coated surfaces.

The CaptureSelect™ Human Fab-kappa Kinetics Biotin Conjugate allows you to:

Screen Fab-antigen interactions—the Human Fab-kappa Kinetics Biotin Conjugate allows high affinity capture of human IgG Fab fragments that possess a kappa light chain. Extreme low off-rates [k(diss)] are achieved through avidity by targeting the CL-kappa domain of the Fab (light chain) and its IgG CH1 domain (heavy chain) simultaneously, thereby enabling Fab screening on target binding kinetics.

Applications for CaptureSelect™ Human Fab-kappa Kinetics Biotin Conjugate include ELISA, Western blot, Gyrolab™-based immunoassays, and label-free detection platforms such as those based on surface plasmon resonance (Biacore™ and IBIS-MX96 systems) and bio-layer interferometry (ForteBio™ Octet™ systems).

Dexamethasone Fluorescein (Invitrogen™)

The synthetic steroid hormone dexamethasone binds to the glucocorticoid receptor, producing a steroid-receptor complex that then localizes in the nucleus and regulates gene transcription. The green fluorescent analog, fluorescein dexamethasone should be useful for studying the mechanism of glucocorticoid receptor activation, although at a slower rate than unmodified dexamethasone.

FL - SRC 3-1 Peptide, 100µM

LanthaScreen® fluorescent nuclear receptor coregulator peptides contain known interaction motifs and are labeled with fluorescein. These peptides are matched and validated to complement the LanthaScreen® TR-FRET Nuclear Receptor Coregulator Assays. Assays developed using these reagents enable primary or secondary screening of nuclear receptor agonists and/or antagonists.

How they work
Ligand binding to nuclear receptors causes conformational changes in the receptor, resulting in a cascade of events, including dissociation of repressor proteins, association of coactivator proteins, and assembly of pol II and other transcriptional factors for activation of target genes. TR-FRET based assays can be developed using the LanthaScreen® panel of fluorescein-labeled coregulator peptides to investigate conformational changes of nuclear receptors upon ligand binding, either by determining the affinity of ligand-bound receptor for different coregulator peptides, or by identifying additional agonists or antagonists via displacement or recruitment of a specific coregulator peptide. In the LanthaScreen® TR-FRET RXR beta Coactivator Assay, a terbium (Tb)-labeled anti-GST antibody is used to indirectly label GSTtagged RXR beta protein. An agonist (9-cis Retinoic Acid) added to the receptor which, upon ligand binding, causes a conformational change resulting in recruitment of the Fl-D22 coactivator peptide (Figure 1). The binding of the fluorescent coregulator peptide to RXR beta causes an increase in the TR-FRET emission ratio (Figure 2).

Contents and Storage:

LanthaScreen® fluorescent nuclear receptor coregulator peptides are supplied as 100 µM solutions in peptide-appropriate buffers. Store peptides at -20°C.

Concanavalin A, Alexa Fluor™ 647 Conjugate (Invitrogen™)

Concanavalin A (Con A) is one of the most widely used lectins in cell biology. Our Alexa Fluor® 647 conjugate of Con A exhibits the bright, far-red fluorescence of the Alexa Fluor® 647 dye (absorption/emission maxima ~650/668 nm). Alexa Fluor® 647 Con A selectively binds to a-mannopyranosyl and a-glucopyranosyl residues.

LanthaScreen™ Tb-Ubiquitin

Ubiquitin has been labeled with the LanthaScreen® Terbium chelate for use in high throughput assays to monitor protein ubiquitination. Due to the method used to label the ubiquitin protein, all of the lysines with the protein are available for further conjugation to form poly-ubiquitin chains.

For use in combination with FRET acceptors including Fluorescein-ubiquitin (PV4377) or a fluorescein-labeled antibody.

BOCILLIN™ FL Penicillin, Sodium Salt (Invitrogen™)

Penicillin-binding proteins, present on the cytoplasmic membranes of eubacteria, can be detected with our BOCILLIN FL penicillin and BOCILLIN 650/665 penicillin (B-13234). BOCILLIN FL penicillin, synthesized from penicillin V and BODIPY FL dye (which is spectrally similar to fluorescein), has been used to determine the penicillin-binding protein profiles of a number of different bacteria types.

FL - TB3 Peptide, 100µM

LanthaScreen® fluorescent nuclear receptor coregulator peptides contain known interaction motifs and are labeled with fluorescein. These peptides are matched and validated to complement the LanthaScreen® TR-FRET Nuclear Receptor Coregulator Assays. Assays developed using these reagents enable primary or secondary screening of nuclear receptor agonists and/or antagonists.

How they work
Ligand binding to nuclear receptors causes conformational changes in the receptor, resulting in a cascade of events, including dissociation of repressor proteins, association of coactivator proteins, and assembly of pol II and other transcriptional factors for activation of target genes. TR-FRET based assays can be developed using the LanthaScreen® panel of fluorescein-labeled coregulator peptides to investigate conformational changes of nuclear receptors upon ligand binding, either by determining the affinity of ligand-bound receptor for different coregulator peptides, or by identifying additional agonists or antagonists via displacement or recruitment of a specific coregulator peptide. In the LanthaScreen® TR-FRET RXR beta Coactivator Assay, a terbium (Tb)-labeled anti-GST antibody is used to indirectly label GSTtagged RXR beta protein. An agonist (9-cis Retinoic Acid) added to the receptor which, upon ligand binding, causes a conformational change resulting in recruitment of the Fl-D22 coactivator peptide (Figure 1). The binding of the fluorescent coregulator peptide to RXR beta causes an increase in the TR-FRET emission ratio (Figure 2).

Contents and Storage:

LanthaScreen® fluorescent nuclear receptor coregulator peptides are supplied as 100 µM solutions in peptide-appropriate buffers. Store peptides at -20°C.