secondary antibody

Goat Anti-Mouse IgG (H+L) Secondary Antibody, TRITC Conjugate

BA1089

0.5 ml

Enabled

Goat Anti-Mouse IgG (H+L) Secondary Antibody, TRITC Conjugate

Secondary Antibodies, IHC ICC IF Antibodies

0.5ml / $135 1ml / $280

Polyclonal

1mg/ml

TRITC

TRITC Conjugated Goat Anti-mouse IgG (H+L) secondary antibody This TRITC conjugated antibody is specific for mouse IgG and shows no cross-reactivity with human/bovine/rabbit IgG.

This antibody is purified from antiserum by immunoaffinity chromatography which removes essentially all goat serum proteins, except the specific antibody for mouse IgG. The antibody preparation is solid phase adsorbed with human serum proteins to ensure minimal cross reactivity in tissue or cell preparations.

Product Overview Product Name Goat Anti-Mouse IgG (H+L) Secondary Antibody, TRITC Conjugate Synonyms TRITC-conjugated Goat Anti-Mouse IgG; Goat Anti-Mouse IgG-TRITC Secondary Antibody; Rhodamine-labeled Goat Anti-Mouse IgG Secondary Antibody Description Goat Anti-Mouse IgG (H+L) Secondary Antibody, TRITC Conjugate, for detection, localization and quantification of target proteins in a sample via indirect immunofluorescence in IHC-P, IHC-F or ICC Reagent Type Fluorophore-conjugated secondary antibody Conjugate TRITC Host Goat Target Species Mouse Antibody Class IgG Clonality Polyclonal Immunogen Whole molecule mouse IgG Purification Immunoaffinity chromatography Specificity Mouse IgG specific Form Supplied Liquid: concentrated buffered stock solution Formulation 0.5 mg TRITC-conjugated secondary antibody . 0.01 M PBS (PH 7.4) . 0.01% Thimerosal . 50% glycerol Pack Size 0.5 ml Concentration 0.1 mg/ml Application Immunohistochemistry (IHC-P and IHC-F), Immunocytochemistry (ICC) . Our Boster Guarantee covers the use of this product in the above marked tested applications. Storage 4C for 1 year Precautions FOR RESEARCH USE ONLY. NOT FOR DIAGNOSTIC OR CLINICAL USE Assay Information Sample Type Mouse primary-antibody-probed formalin-fixed paraffin-embedded (FFPE) tissue sections on slides (IHC-P), or thawed frozen samples (IHC-F) Assay Type Immunoanalytical Technique Indirect immunofluorescence Assay Purpose Protein detection/quantification Equipment Needed Excitation light source, filter set and detector, fluorescence microscope (can be combined with confocal microscope), fluorescence plate-reader, flow cytometer, or cell sorter Main Advantages Specific High signal-to-noise ratio High Signal Amplification Multiple secondary antibodies can bind to a single primary antibody, multiple TRITC molecules bind to a single secondary antibody Fast Fewer processing steps - no need to add a substrate; Less optimization required compared to enzymatic detection; Generates strong signals in a relatively short time span; signal can be observed directly Quantifieable The digital nature of the gold signal + high precision in allocating gold labels to defined structures makes it easy to count and quantify Flexible No need to label each antibody against each target protein with a fluorescent dye, the small size of TRITC causes no steric interference with proper biological function of target proteins or antibodies Multiplex Compatible Compatible with colocalization studies (multiple antigens concurrent detection) even in close proximity using primary antibodies from different host species for simultaneous detection by fluorophore-conjugated secondary antibodies, or using multiple differently colored fluorophores (FITC and TRITC) in the same experiment for target differentiation. Background . Most commonly, secondary antibodies are generated by immunizing the host animal with a pooled population of immunoglobulins from the target species. The host antiserum is then purified through immunoaffinity chromatography to remove all host serum proteins, except the specific antibody of interest, and then modified with antibody fragmentation, label conjugation, etc., to generate highly specific detection reagents. Secondary antibodies can be conjugated to a large number of labels, including enzymes, biotin, fluorescent dyes/proteins, or gold particles. Here, the antibody provides the specificity to locate the protein of interest by recognizing a primary antibody that targets a particular antigen, and the label generates a detectable signal in the area of the formed immune complexes. The label of choice depends upon the experimental application. . Fluorescent reporters widely used in biological research are of two types: organic compounds with a low molecular weight (0.2-1 kDa) typically containing numerous aromatic groups, or plane or cyclic moieties with π bonds (e.g.FITC, TRITC, Alexa Fluor Dyes, DyLight Fluors), and biological fluorophores (e.g.green fluorescent protein (GFP), R-Phycoerythrin). TRITC (tetramethylrhodamine isothiocyanate )is a bright orange-fluorescent dye. It is a derivative of rhodamine (a member of the fluorone dyes family) where a hydrogen atom on the bottom ring of the structure is replaced by isothiocyanate functional group (-N=C=S), making it more reactive to amine and sulfhydryl groups on proteins. The bond to antibodies is based on this reactive group. The excitation and emission wavelengths of TRITC are 550 nm and 573 nm respectively. The optimal degree of conjugation for least changes in the antibody affinity and maximal specific staining (fluorescence) is at a molecular ratio TRITC/Ab of approximately 2. Like most fluorochromes, TRITC is prone to photobleaching, i.e. losing fluorescing properties due to molecule structure degradation. TRITC is inherently a quite stable dye, and thus exhibits less photobleaching than fluorescein, which is commonly used as a photobleaching standard.Loss of activity caused by photobleaching can be controlled by reducing the intensity or time-span of light exposure, by increasing the concentration of the fluorophore, or by employing more robust fluorophores that are less prone to bleaching (e.g., Alexa Fluors, Seta Fluors, or DyLight Fluors). Analogs of TRITC with greater photostability and higher fluorescence intensity tailored in various biological applications are Alexa 555 and DyLight 550.

0.5 ml

Goat

Mouse IgG (whole molecule).

At 4 C for one year.

IHC, ICC

Mouse

Figure 1. IHC analysis of rat heart using anti-Alpha-Smooth Muscle Actin antibody (MA1106). Alpha-Smooth Muscle Actin was detected in paraffin-embedded section of rat heart tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1ug/ml mouse anti- Alpha-Smooth Muscle Actin Antibody (MA1106) overnight at 4°C. TRITC Conjugated goat anti-mouse IgG (BA1089) was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1021) with DAB as the chromogen.

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