Rabbit Anti-Human IgG (H+L) Secondary Antibody, Cy3 Conjugated

This antibody is purified from antiserum by immunoaffinity chromatography which removes essentially all rabbit serum proteins, except the specific antibody for human IgG. Cited in 7 publication(s).

Product Info Summary

SKU: BA1035
Size: 0.5ml
Host: Rabbit
Application: Flow Cytometry, IF

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Product Overview

Product Name Rabbit Anti-Human IgG (H+L) Secondary Antibody, Cy3 Conjugate
Synonyms Cy3-conjugated Rabbit Anti-Human IgG; Rabbit Anti-Human IgG-Cyanine 3 Secondary Antibody; Cy3-labeled Rabbit Anti-Human IgG Secondary Antibody
Description Rabbit Anti-Human IgG (H+L) Secondary Antibody, Cy3 Conjugate, for detection, localization and quantification of target proteins in sliced samples via indirect immunofluorescence in IHC-P, IHC-F or ICC.
Reagent Type Fluorophore-conjugated secondary antibody
Conjugate Cy3
Host Rabbit
Target Species Human
Antibody Class IgG
Clonality Polyclonal
Immunogen Whole molecule human IgG
Purification Immunoaffinity chromatography
Specificity Human IgG specific
Form Supplied Liquid: concentrated buffered stock solution
Formulation 0.5 mg Cy3-conjugated secondary antibody
0.01 M PBS (PH 7.4)
5 mg/mL BSA
50% glycerol
Pack Size 0.5 ml
Concentration 0.1 mg/ml
Application IF, Flow Cytometry
Our Boster Guarantee covers the use of this product in the above marked tested applications.
Storage At -20˚C for one year from date of receipt. Avoid repeated freezing and thawing.
Precautions FOR RESEARCH USE ONLY. NOT FOR DIAGNOSTIC OR CLINICAL USE

Assay Information

Sample Type Human 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

Main Advantages

Specific High signal-to-noise ratio
High Signal Amplification Multiple secondary antibodies can bind to a single primary antibody, multiple Cy3 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
Absolute Quantification Precise quantitative analysis of fluorescence microscopy images can provide absolute protein amount and information regarding stoichiometry of protein complexes
Stability -Cyanine dyes are better able to withstand the harsh dehydration and embedding conditions required for mounting sections
-Reduced fluorescence quenching (sulfonated cyanines are less prone to aggregation in water due to reduced dye-dye interactions)
-Cy3 conjugates are bright orange-red and more photostable in the aqueous environment, as well as in non-polar media
Superb Signal Detection Precise target localization, high extinction coefficient, and good quantum yield of Cy3 results in brighter fluorescence and less background than other orange-red fluorescing dye conjugates
Flexible No need to label each antibody against each target protein with a fluorescent dye
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). Cy3 is a cyanine dye with 3 carbon atoms/methine groups between two indolenine groups, NHS-ester, sulfonated at both indoles. It fluoresces greenish yellow with excitation max at ~550 nm and emission at ~570 nm. Cy3 can be detected by various fluorometers, imagers, and microscopes with standard filters for Tetramethylrhodamine (TRITC). Due to inherently high extinction coefficient, this dye is also easily detected by naked eye on gels, and in solution. Cy3 can be excited to about 50% of maximum with an argon laser (514 nm or 528 nm lines), or to about 75% of maximum with a helium/neon laser (543 nm line) or mercury lamp (546 nm line). For double labeling Cy3 has been typically used with Cy5, with fluorescein (using narrow band-pass emission filter to minimize Cy3 fluorescence in the FITC filter set), and also with with Alexa Fluor 647 for multiple labeling when using a confocal microscope. Analog of Cy3 with greater photostability and higher fluorescence intensity suited for various biological applications such as imaging and flow cytometry, is Alexa Fluor 555 dye.

Validation Images & Assay Conditions

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BA1035 has been cited in 7 publications:

*The publications in this section are manually curated by our staff scientists. They may differ from Bioz's machine gathered results. Both are accurate. If you find a publication citing this product but is missing from this list, please let us know we will issue you a thank-you coupon.

Honokiol Attenuates Oligomeric Amyloid β1-42-Induced Alzheimer’s Disease in Mice Through Attenuating Mitochondrial Apoptosis and Inhibiting the Nuclear Factor Kappa-B Signaling Pathway

Design of magnetic nanoparticles for hepatocellular carcinoma treatment using the control mechanisms of the cell internal nucleus and external membrane

Penton-dodecahedron of fowl adenovirus serotype 4 as a vaccine candidate for the control of related diseases

Aldehyde oxidase 1 promoted the occurrence and development of colorectal cancer by up-regulation of expression of CD133

Yin Z,Hua L,Chen L,Hu D,Li J,An Z,Tian T,Ning H,Ge Y.Bisphenol-A exposure induced neurotoxicity and associated with synapse and cytoskeleton in Neuro-2a cells.Toxicol In Vitro.2020 Sep;67:104911.doi:10.1016/j.tiv.2020.104911.Epub 2020 Jun 5.PMID:32512148.
Species: Mouse
BA1035 usage in article: APP:IF, SAMPLE:NEURO-2A CELL, DILUTION:NA

Zhang W,Chai W,Zhu Z,Li X.Aldehyde oxidase 1 promoted the occurrence and development of colorectal cancer by up-regulation of expression of CD133. Int Immunopharmacol.2020 Aug;85:106618.doi:10.1016/j. intimp.2020.106618.Epub 2020 May 26.PMID:32470878.
Species: Human
BA1035 usage in article: APP:IF, SAMPLE:LOVO CELLS, DILUTION:1:400

Olfactory ensheathing cells promote nerve regeneration and functional recovery after facial nerve defects Jian Gu,1,et al. Neural Regen Res. 2019 Jan; 14(1): 124–131. doi: 10.4103/1673-5374.243717

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1 Customer Q&As for Rabbit Anti-Human IgG (H+L) Secondary Antibody, Cy3 Conjugated

Question

Could you confirm whether the quality of BA1035 is okay to store in -20C for 2-3 days?

Verified customer

Asked: 2019-06-09

Answer

For the Rabbit Anti-Human IgG (H+L) Secondary Antibody, Cy3 Conjugate (BA1035), there should not be any quality issues if they were stored at -20°C for a few days.

Boster Scientific Support

Answered: 2019-06-10

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