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Goat anti rabbit conjugated to cy3

Manufactured by Jackson ImmunoResearch
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Goat anti-rabbit conjugated to Cy3 is a secondary antibody used in immunological applications. It is produced by immunizing goats with rabbit immunoglobulins and then conjugating the resulting antibodies to the fluorescent dye Cy3. This product can be used to detect and visualize the presence of rabbit primary antibodies in various experimental techniques.

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Lab products found in correlation

Vectashield mounting media, by Vector Laboratories (1 mentions) Axioplan 2 microscope, by Zeiss (1 mentions) Orca 2 camera, by Hamamatsu Photonics (1 mentions) Dylight 488, by Vector Laboratories (1 mentions) Alexa fluor 647, by Abcam (1 mentions) Anti gfap, by Merck Group (1 mentions) Goat anti mouse igg h l, by Abcam (1 mentions) Mounting medium, by Thermo Fisher Scientific (1 mentions) Goat anti mouse conjugated to alexa fluor 488, by Thermo Fisher Scientific (1 mentions) Rabbit anti myc, by Abcam (1 mentions) 4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (1 mentions) Mouse anti flag, by Merck Group (1 mentions) Mouse anti ha, by Fortrea (1 mentions) Goat anti rabbit antibodies conjugated to alexa fluor 488, by Thermo Fisher Scientific (1 mentions)

Close spelling variants of this product

Goat anti-rabbit Cy3 Cy3 anti-goat Anti-rabbit Cy3

5 protocols using goat anti rabbit conjugated to cy3

1

Kinetochore distance measurement protocol

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Cells grown on 22-mm2 coverslips were simultaneously fixed and permeablized with 2% paraformaldehyde and 0.5% Triton X-100 in 1X PHEM buffer at room temperature for 15 min. The cells were blocked with 20% boiled normal goat serum (BNGS) for at least 20 min. Coverslips were incubated with primary antibody, Anti-­centromere antibody (1:800, Antibody Inc., 15-134), and rabbit anti SGO1 (1:500, a gift from Hongtao Yu, University of Texas Southwestern Medical Center) diluted in 5% BNGS in PBST overnight at 4°C. Coverslips were washed three times with MOPS buffered saline with 0.05% Tween 20 and then incubated in secondary antibody, goat anti-rabbit conjugated to CY3 at 1:1500 (JacksonImmuno­Research, 111-165-045109), and goat anti-human conjugated to fluorescein isothiocyanate at 1:800 (JacksonImmunoResearch, 109-95-088) for 2 h at room temperature. After incubation with secondary antibodies, coverslips were washed three times again and then labeled with DAPI (100 ng/ml) for 1 min. Coverslips were mounted on slides with Vectashield mounting media (Vector Laboratories, H-1000) and then sealed with clear nail polish. Fluorescence images of cells were taken using a Zeiss Axioplan II microscope with a Zeiss 100× objective, Hamamatsu Orca II camera, and Metamorph software. Distances between pairs of kinetochores were measured using the region measurement tool in Metamorph software.
Sapkota H., Wasiak E., Daum J.R, & Gorbsky G.J. (2018). Multiple determinants and consequences of cohesion fatigue in mammalian cells. Molecular Biology of the Cell, 29(15), 1811-1824.
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2

Multicolor Immunofluorescence Labeling

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Goat anti-rabbit conjugated to Cy3 (1:500; Jackson ImmunoResearch Labs, West Grove, PA, United States, Cat #111-165-003, RRID:AB_2338000), goat anti-rabbit conjugated to Alexa fluor 488 (1:500; Thermo Fisher Scientific, Waltham, MA, United States, Cat #A-11034, RRID:AB_2576217), goat anti chicken conjugated to Alexa fluor 647 (1:500; Thermo Fisher Scientific, Waltham, MA, United States, Cat #A21449, RRID:Ab_2535866), goat anti-chicken conjugated to Alexa fluor 488 (1:500; Thermo Fisher Scientific, Waltham, MA, United States, Cat #A11039, RRID:AB_2534096), goat anti-mouse conjugated to Cy3 (1:500, Jackson ImmunoResearch Labs, West Grove, PA, United States, Cat #115-165-003, RRID:AB_2338680), goat anti-guinea pig conjugated to Cy3 (1:500; Jackson ImmunoResearch Labs, West Grove, PA, United States, Cat #106-165-003, RRID:AB_2337398) were used.
Ferdos S., Brockhaus J., Missler M, & Rohlmann A. (2022). Deletion of β-Neurexins in Mice Alters the Distribution of Dense-Core Vesicles in Presynapses of Hippocampal and Cerebellar Neurons. Frontiers in Neuroanatomy, 15, 757017.
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3

Immunolabeling of Dystrophin-Associated Proteins

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The rabbit polyclonal IgG antibodies were: H4 directed against exons 78-79 of the C-terminus dystrophin (1:400), LG5 anti-β-dystroglycan (1:400) and K7 anti-utrophin (1:200) (gift from D. Mornet, Montpellier), anti-Aquaporin-4 (AQP4 300-314; 1:400; Alomone labs, Israël), anti-AQP4ex (60789S, 1:400, Ozyme, France), anti-alpha-1 syntrophin (1:200; Alomone labs, Israël), anti-alpha smooth muscle actin (anti-SMA; 1:200; Sigma) and anti-Glial Fibrillary Acidic Protein (Anti-GFAP; 1:1000, Dako, Denmark). The mouse monoclonal antibodies were: Anti-GFAP (1:500, Sigma-Aldrich), sc-390488 anti-AQP4 (1:50 in westernblots; Santa Cruz Biotechnology) and anti-Dp427 N-terminal NCL-Dys1 (diluted 1:3; Leica Biosystem).
The secondary antibodies were: goat anti-rabbit conjugated to Cy3 (diluted 1:400, Jackson Immunoresearch, USA) and goat anti-mouse IgG H&L (diluted 1:400, Alexa Fluor® 647, Abcam). The Lycopersicon esculentum lectin Dy-Light 488 (1:100, Vector laboratories), specific of N-acetyl-D-glucosamin and N-acetyl-polylactosamine oligomers (Kawashima, Sueyoshi, Li, Yamamoto, & Osawa, 1990; (link)Porter, Palade, & Milici, 1990) , was used as an endothelial marker to label the vessels and as a control for normalization of immunofluorescence quantifications.
Belmaati Cherkaoui M., Vacca O., Izabelle C., Boulay A.C., Boulogne C., Gillet C., Barnier J.V., Rendon A., Cohen-Salmon M, & Vaillend C. (2021). Dp71 contribution to the molecular scaffold anchoring aquaporine-4 channels in brain macroglial cells. Glia, 69(4).
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4

Subcellular Localization of Transfected Proteins

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The subcellular distribution of polymeric proteins was assessed in transfected HEK293T cells by immunofluorescence. Cells were plated on 8 well tissue-culture chambers and transfected with plasmids the next day. Forty-eight hours post-transfection, cells were fixed in 4% paraformaldehyde (PFA) in PBS for 30 min and permeabilized in 0.5% triton X-100 in PBS for 15 min on ice. The cells were blocked in 1% normal goat serum (NGS) in PBS for 30 min. After blocking, the cells were incubated for 1 hour at RT in blocking solution containing the rabbit anti-Myc (Abcam), mouse anti-HA (Covance), mouse anti-Flag (Sigma), rabbit α-LPAC primary antibodies at a dilution of 1:400. The slides were washed three times in PBS and incubated for 1 hour at RT in blocking solution containing Goat anti-rabbit conjugated to Cy3 (Jackson ImmunoResearch, PA) and goat anti-mouse conjugated to Alexa Fluor 488 (Invitrogen) secondary antibodies at a dilution of 1:200. The slides were washed three times in PBS and mounted with mounting medium containing DAPI (Invitrogen).
Zu T., Cleary J.D., Liu Y., Bañez-Coronel M., Bubenik J.L., Ayhan F., Ashizawa T., Xia G., Clark H.B., Yachnis A.T., Swanson M.S, & Ranum L.P. (2017). RAN Translation Regulated by Muscleblind Proteins in Myotonic Dystrophy Type 2. Neuron, 95(6), 1292-1305.e5.
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5

Immunohistochemistry of Neuromodulators

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Tissues were next incubated in primary antibody solutions for one week at 4 °C. Antibodies raised in rabbits against ELH were obtained from J. Blankenship (University of Texas), while rabbit antibodies against αCDCP, APGWamide and MIP were all obtained from J. van Minnen (Vrije Universiteit of Amsterdam). References describing the antigens used for raising each antibody are provided in Table 1. Finally, antibodies raised in rabbits against FMRFamide were obtained commercially from ImmunoStar (Hudson, WI USA, Cat #20091). The dilutions of each of the primary antibodies are also given in Table 1.
Following primary incubation, the ganglia were washed four times over 8-12 hours in fresh blocking solution. Tissue was then incubated for 5 days at 4 °C in darkness in either goat-antirabbit antibodies conjugated to AlexaFluor488 (Molecular Probes, Eugene, OR, USA; Cat # A11008) or goat-anti-rabbit conjugated to Cy3 (Jackson ImmunoResearch, West Grove, PA, USA; Cat #111-165-003), both diluted 1:200 in blocking solution. The ganglia were then washed for 6-8 hours in blocking solution before being transferred to glycerol mounting medium (25% 0.05M Tris buffer in 75% glycerol with the addition of 2% n-propyl-gallate to minimize photobleaching).
Acker M.J., Habib M.R., Beach G.A., Doyle J.M., Miller M.W, & Croll R.P. (2019). An immunohistochemical analysis of peptidergic neurons apparently associated with reproduction and growth in Biomphalaria alexandrina. General and comparative endocrinology, 280, 1-8.
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