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SKU BA1032
Pack size 0.5 ml
Applications IHC-P, IHC-F, ICC

Product Overview

Product Name Goat Anti-Rabbit IgG (H+L) Secondary Antibody, Cy3 Conjugate
Synonyms Cy3-conjugated Goat Anti-Rabbit IgG; Goat Anti-Rabbit IgG-Cyanine 3 Secondary Antibody; Cy3-labeled Goat Anti-Rabbit IgG Secondary Antibody
Description Goat Anti-Rabbit 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 Goat
Target Species Rabbit
Antibody Class IgG
Clonality Polyclonal
Immunogen Whole molecule rabbit IgG
Purification Immunoaffinity chromatography
Specificity Rabbit IgG specific
Form Supplied Liquid: concentrated buffered stock solution
Formulation 0.5 mg Cy3-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

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.

Goat Anti-Rabbit IgG (H+L) Secondary Antibody, Cy3 Conjugate Images

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Goat Anti-Rabbit IgG (H+L) Secondary Antibody, Cy3 Conjugate
MCM2 was detected in paraffin-embedded sections of human intestinal cancer tissues using rabbit anti- MCM2 Antigen Affinity purified polyclonal antibody (Catalog # PA1650) at 1 μg/mL. Cy3 Conjugated Goat Anti-Rabbit IgG (H+L) secondary antibody (Catalog # BA1032) was used to detect the primary antibody at 20μg/mL.
Goat Anti-Rabbit IgG (H+L) Secondary Antibody, Cy3 Conjugate
Collagen IV was detected in paraffin-embedded sections of rat kidney tissues using rabbit anti- Collagen IV Antigen Affinity purified polyclonal antibody (Catalog # PA1521) at 1 ??g/mL. Cy3 Conjugated Goat Anti-Rabbit IgG (H+L) secondary antibody (Catalog # BA1032) was used to detect the primary antibody at 20??g/mL.
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Publications

Enhancer of zeste homolog 2 silencing inhibits tumor growth and lung metastasis in osteosarcoma
EZH2 promotes DNA replication by stabilizing interaction of POL? and PCNA via methylation-mediated PCNA trimerization
Manipulation of autophagy: a novelly potential therapeutic strategy for retinal neovascularization
Autophagy activation and the mechanism of retinal microvascular endothelial cells in hypoxia
Icariin Regulates the Bidirectional Differentiation of Bone Marrow Mesenchymal Stem Cells through Canonical Wnt Signaling Pathway
Regulatory role of NKG2D+ NK cells in intestinal lamina propria by secreting double-edged Th1 cytokines in ulcerative colitis
Protection by simvastatin on hyperglycemia-induced endothelial dysfunction through inhibiting NLRP3 inflammasomes
Inhibition of HMGB1 reduces rat spinal cord astrocytic swelling and AQP4 expression after oxygen-glucose deprivation and reoxygenation via TLR4 and NF-?B signaling in an IL-6-dependent manner
Expression of MicroRNA-29a Regulated by Yes-Associated Protein Modulates the Neurite Outgrowth in N2a Cells
Phosphoinositide-specific phospholipase C?1 inhibition induces autophagy in human colon cancer and hepatocellular carcinoma cells
Solasodine inhibits human colorectal cancer cells through suppression of the AKT/glycogen synthase kinase?3?/??catenin pathway
The novel cannabinoid receptor GPR55 mediates anxiolytic-like effects in the medial orbital cortex of mice with acute stress
Peroxisome proliferator-activated receptor ? mediates porcine placental angiogenesis through hypoxia inducible factor-, vascular endothelial growth factor- and angiopoietin-mediated signaling
Expression, Localization of SUMO-1, and Analyses of Potential SUMOylated Proteins in Bubalus bubalis Spermatozoa
West Nile Virus NS1 Antagonizes Interferon Beta Production by Targeting RIG-I and MDA5
Neuroprotective effect of combining tanshinone IIA with low-dose methylprednisolone following acute spinal cord injury in rats
Interleukin 22 attenuated angiotensin II induced acute lung injury through inhibiting the apoptosis of pulmonary microvascular endothelial cells
Sodium tanshinone IIA sulfonate improves inflammation, aortic endothelial cell apoptosis, disseminated intravascular coagulation and multiple organ damage in a rat heat stroke model
Nitric Oxide Synthase Activity Correlates with OGG1 in Ozone-Induced Lung Injury Animal Models
Human cytomegalovirus immediate-early protein promotes survival of glioma cells through interacting and acetylating ATF5
Simultaneous exposure to Fc?R and Fc?R on monocytes and macrophages enhances antitumor activity in vivo
Zonisamide-loaded triblock copolymer nanomicelles as a novel drug delivery system for the treatment of acute spinal cord injury
Abce1 orchestrates M-phase entry and cytoskeleton architecture in mouse oocyte
miR-1181 inhibits invasion and proliferation via STAT3 in pancreatic cancer
Olfactory Ensheathing Cells Inhibit Gliosis in Retinal Degeneration by Downregulation of the M?ller Cell Notch Signaling Pathway
Astaxanthin protects against early burn-wound progression in rats by attenuating oxidative stress-induced inflammation and mitochondria-related apoptosis
Autophagy protects gastric mucosal epithelial cells from ethanol-induced oxidative damage via mTOR signaling pathway
Salidroside promotes peripheral nerve regeneration based on tissue engineering strategy using Schwann cells and PLGA: in vitro and in vivo
Cellular Localization and Regulation of Expression of the PLET1 Gene in Porcine Placenta
Adiponectin reduces ER stress-induced apoptosis through PPAR? transcriptional regulation of ATF2 in mouse adipose
Thioredoxin-Interacting Protein Mediates NLRP3 Inflammasome Activation Involved in the Susceptibility to Ischemic Acute Kidney Injury in Diabetes
VE-cadherin involved in the pulmonary microvascular endothelial cell barrier injury induced by angiotensin II through modulating the cellular apoptosis and skeletal rearrangement
A novel co-drug of aspirin and ursolic acid interrupts adhesion, invasion and migration of cancer cells to vascular endothelium via regulating EMT and EGFR-mediated signaling pathways: multiple targets for cancer metastasis prevention and treatment
Imbalance between Glutamate and GABA in Fmr1 Knockout Astrocytes Influences Neuronal Development
SDF?1/CXCR4 axis induces apoptosis of human degenerative nucleus pulposus cells via the NF??B pathway
Angiotensin II is related to the acute aortic dissection complicated with lung injury through mediating the release of MMP9 from macrophages
Astragalus membranaceus?ameliorates renal interstitial fibrosis by inhibiting tubular epithelial-mesenchymal transition?in vivo?and?in vitro
SIV Infection Impairs the Central Nervous System in Chinese Rhesus Macaques
Controlled water vapor transmission rate promotes wound-healing via wound re-epithelialization and contraction enhancement
The GTPase SPAG-1 orchestrates meiotic program by dictating meiotic resumption and cytoskeleton architecture in mouse oocytes
SIRT3 Deficiency Induces Endothelial Insulin Resistance and Blunts Endothelial-Dependent Vasorelaxation in Mice and Human with Obesity
Pharmacoproteomic analysis reveals that metapristone (RU486 metabolite) intervenes E-cadherin and vimentin to realize cancer metastasis chemoprevention
Murine Sertoli cells promote the development of tolerogenic dendritic cells: a pivotal role of galectin-1
Killing of Staphylococcus aureus via Magnetic Hyperthermia Mediated by Magnetotactic Bacteria
Difference in expression patterns of placental cholesterol transporters, ABCA1 and SR-BI, in Meishan and Yorkshire pigs with different placental efficiency
Overexpression of transcription factor Foxa2 and Hnf1? induced rat bone mesenchymal stem cells into hepatocytes
Salidroside protects against bleomycin-induced pulmonary fibrosis: activation of Nrf2-antioxidant signaling, and inhibition of NF-?B and TGF-?1/Smad-2/-3 pathways
Angiotensin II down-regulates nephrin-Akt signaling and induces podocyte injury: roleof c-Abl
Microencapsulation improves inhibitory effects of transplanted olfactory ensheathing cells on pain after sciatic nerve injury
Nanog down-regulates the Wnt signaling pathway via ?-catenin phosphorylation during epidermal stem cell proliferation and differentiation
Synergistic effects of Buyang Huanwu decoction and embryonic neural stem cell transplantation on the recovery of neurological function in a rat model of spinal cord injury
Gastrodin blocks neural stem cell differentiation into glial cells mediated by kainic acid
Favorable proliferation and differentiation capabilities of neural precursor cells derived from rat cochlear nucleus
Isolation, cultivation and identification of human lung adenocarcinoma stem cells
Telocytes damage in endometriosis-affected rat oviduct and potential impact on fertility
Chloride channel blocker 4,4-diisothiocyanatostilbene-2,2?-disulfonic acid inhibits nitric oxide-induced apoptosis in cultured rat hippocampal neurons
Injury of cortical neurons is caused by the advanced glycation end products-mediated pathway
The Effect of Electroacupuncture on the Extracellular Matrix Synthesis and Degradation in a Rabbit Model of Disc Degeneration
Ultrastructural Localization of Polygalacturonase in Ethylene-Stimulated Abscission of Tomato Pedicel Explants
Investigation of Characteristics of Feather Follicle Stem Cells and Their Regeneration Potential
The Spatiotemporal Development of Intercalated Disk in Three-Dimensional Engineered Heart Tissues Based on Collagen/Matrigel Matrix
Identification of U251 glioma stem cells and their heterogeneous stem-like phenotypes
Transcriptome Sequencing Reveals Differences between Primary and Secondary Hair Follicle-derived Dermal Papilla Cells of the Cashmere Goat (Capra hircus)
SUMOylation of Mouse p53b by SUMO-1 Promotes Its Pro-Apoptotic Function in Ovarian Granulosa Cells
Excessive Astrocyte-Derived Neurotrophin-3 Contributes to the Abnormal Neuronal Dendritic Development in a Mouse Model of Fragile X Syndrome
Characterization and retinal neuron differentiation of WERI-Rb1 cancer stem cells
Nanog and ?-catenin: A new convergence point in EpSC proliferation and differentiation
NDRG2 Ameliorates Hepatic Fibrosis by Inhibiting the TGF-?1/Smad Pathway and Altering the MMP2/TIMP2 Ratio in Rats
Role of bone morphogenetic proteins in form-deprivation myopia sclera
Sphingolipids are involved in?N-methyl-N?-nitro-N-nitrosoguanidine-induced epidermal growth factor receptor clustering
Expression and significance of hypoxia-inducible factor-1 alpha and MDR1/P-glycoprotein in human colon carcinoma tissue and cells
Catalpol Increases Brain Angiogenesis and Up-Regulates VEGF and EPO in the Rat after Permanent Middle Cerebral Artery Occlusion
Gene expression and differentiation characteristics in mice E13.5 and E17.5 neural retinal progenitors
Nanog and ?-catenin: A new convergence point in EpSC proliferation and differentiation
NDRG2 Ameliorates Hepatic Fibrosis by Inhibiting the TGF-?1/Smad Pathway and Altering the MMP2/TIMP2 Ratio in Rats
Role of bone morphogenetic proteins in form-deprivation myopia sclera
Sphingolipids are involved in
Expression and significance of hypoxia-inducible factor-1 alpha and MDR1/P-glycoprotein in human colon carcinoma tissue and cells
Catalpol Increases Brain Angiogenesis and Up-Regulates VEGF and EPO in the Rat after Permanent Middle Cerebral Artery Occlusion
Gene expression and differentiation characteristics in mice E13.5 and E17.5 neural retinal progenitors
Zhang HL, Ye HQ, Liu SQ, Deng CL, Li XD, Shi PY, Zhang B. J Virol. 2017 Aug 24;91(18). pii: e02396-16. doi: 10.1128/JVI.02396-16. Print 2017 Sep 15. West Nile Virus NS1 Antagonizes Interferon Beta Production by Targeting RIG-I and MDA5
Dai L, Chen X, Lu X, Wang F, Zhan Y, Song G, Hu T, Xia C, Zhang B. Sci Rep. 2017 Oct 24;7(1):13912. doi: 10.1038/s41598-017-13334-y. Phosphoinositide-specific phospholipase Cγ1 inhibition induces autophagy in human colon cancer and hepatocellular carcinoma cells
Li R, Wang LZ, Du JH, Zhao L, Yao Y. Int J Ophthalmol. 2018 Apr 18;11(4):567-574. doi: 10.18240/ijo.2018.04.05. eCollection 2018. Autophagy activation and the mechanism of retinal microvascular endothelial cells in hypoxia
Li R, Tian J, Du J, Zhao L, Yao Y, Yu Z, Chang W, Shi R, Li J. BMC Ophthalmol. 2018 Apr 27;18(1):110. doi: 10.1186/s12886-018-0774-6. Manipulation of autophagy: a novelly potential therapeutic strategy for retinal neovascularization
Chang W, Bai J, Tian S, Ma M, Li W, Yin Y, Deng R, Cui J, Li J, Wang G, Zhang P, Tao K. Exp Biol Med (Maywood). 2017 May;242(10):1025-1033. doi: 10.1177/1535370216686221. Epub 2017 Jan 1. Autophagy protects gastric mucosal epithelial cells from ethanol-induced oxidative damage via mTOR signaling pathway
Li B, Xu L, Tao F, Xie K, Wu Z, Li Y, Li J, Chen K, Pi C, Mendelsohn A, Larrick JW, Gu H, Fang J. Oncotarget. 2017 Jun 13;8(24):39356-39366. doi: 10.18632/oncotarget.17000. Simultaneous exposure to FcγR and FcαR on monocytes and macrophages enhances antitumor activity in vivo
Tan C, Yu C, Song Z, Zou H, Xu X, Liu J. Biomed Res Int. 2017;2017:5251236. doi: 10.1155/2017/5251236. Epub 2017 Sep 12. Expression of MicroRNA-29a Regulated by Yes-Associated Protein Modulates the Neurite Outgrowth in N2a Cells
Wang F, Peng PL, Lin X, Chang Y, Liu J, Zhou R, Nie JY, Dong WG, Zhao Q, Li J. Oncotarget. 2017 Oct 30;8(58):98945-98952. doi: 10.18632/oncotarget.22132. eCollection 2017 Nov 17. Regulatory role of NKG2D+ NK cells in intestinal lamina propria by secreting double-edged Th1 cytokines in ulcerative colitis
Wang J, Guo XJ, Ding YM, Jiang JX. World J Gastroenterol. 2017 Mar 7;23(9):1594-1601. doi: 10.3748/wjg.v23.i9.1594. miR-1181 inhibits invasion and proliferation via STAT3 in pancreatic cancer
Xie J, Huo S, Li Y, Dai J, Xu H, Yin ZQ. Cell Transplant. 2017 Jun 9;26(6):967-982. doi: 10.3727/096368917X694994. Epub 2017 Feb 9. Olfactory Ensheathing Cells Inhibit Gliosis in Retinal Degeneration by Downregulation of the M
Hu M, Wang B, Qian D, Wang M, Huang R, Wei L, Li L, Zhang L, Liu DX. Oncotarget. 2017 May 9;8(19):32157-32170. doi: 10.18632/oncotarget.17150. Human cytomegalovirus immediate-early protein promotes survival of glioma cells through interacting and acetylating ATF5
Huang JM, Bao Y, Xiang W, Jing XZ, Guo JC, Yao XD, Wang R, Guo FJ. Evid Based Complement Alternat Med. 2017;2017:8085325. doi: 10.1155/2017/8085325. Epub 2017 Dec 27. Icariin Regulates the Bidirectional Differentiation of Bone Marrow Mesenchymal Stem Cells through Canonical Wnt Signaling Pathway
Zhang S, Li J, Li Y, Liu Y, Guo H, Xu X. Front Physiol. 2017 Apr 27;8:249. doi: 10.3389/fphys.2017.00249. eCollection 2017. Nitric Oxide Synthase Activity Correlates with OGG1 in Ozone-Induced Lung Injury Animal Models
Chen F, Li H, Zhu G, Chen X, Tang Z. Mol Med Rep. 2017 Jul;16(1):87-94. doi: 10.3892/mmr.2017.6573. Epub 2017 May 11. Sodium tanshinone IIA sulfonate improves inflammation, aortic endothelial cell apoptosis, disseminated intravascular coagulation and multiple organ damage in a rat heat stroke model
Jiao XF, Huang CJ, Wu D, Zhang JY, Long YT, Chen F, Li X, Huo LJ. Oncotarget. 2017 Jun 13;8(24):39012-39020. doi: 10.18632/oncotarget.16546. Abce1 orchestrates M-phase entry and cytoskeleton architecture in mouse oocyte
Li J, Deng J, Yuan J, Fu J, Li X, Tong A, Wang Y, Chen Y, Guo G. Int J Nanomedicine. 2017 Mar 29;12:2443-2456. doi: 10.2147/IJN.S128705. eCollection 2017. Zonisamide-loaded triblock copolymer nanomicelles as a novel drug delivery system for the treatment of acute spinal cord injury
Fang Q, Guo S, Zhou H, Han R, Wu P, Han C. Sci Rep. 2017 Jan 27;7:41440. doi: 10.1038/srep41440. Astaxanthin protects against early burn-wound progression in rats by attenuating oxidative stress-induced inflammation and mitochondria-related apoptosis
Zhang J, Peng X, Yuan A, Xie Y, Yang Q, Xue L. Mol Med Rep. 2017 Sep;16(3):2636-2644. doi: 10.3892/mmr.2017.6903. Epub 2017 Jun 30. Peroxisome proliferator-activated receptor γ mediates porcine placental angiogenesis through hypoxia inducible factor-, vascular endothelial growth factor- and angiopoietin-mediated signaling
Liu H, Lv P, Zhu Y, Wu H, Zhang K, Xu F, Zheng L, Zhao J. Sci Rep. 2017 Jan 5;7:39869. doi: 10.1038/srep39869. Salidroside promotes peripheral nerve regeneration based on tissue engineering strategy using Schwann cells and PLGA: in vitro and in vivo
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Xiong, S., Zhang, S., Guan, L., Chen, J., Tu, X., Li, Q., & Jiang, H. (2017). Differentiation of induced pluripotent stem cells for future olfactory repair using an indirect co-culture technique. International Journal of Clinical and Experimental Pathology, 10(7), 8072-8081. Retrieved from http://www.ijcep.com/files/ijcep0055217.pdf
Han, P., Zhang, R., Gao, Y., Niu, X., Chen, Z., Zhang, Q., & Xu, M. (2017). Expression patterns of fibroblast growth factor 8 in the spiral ganglion of rat cochlea. International Journal of Clinical and Experimental Medicine, 10(6), 9345-9351. Retrieved from http://www.ijcem.com/files/ijcem0050425.pdf
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