Goat anti rabbit igg alexa fluor plus 647

Manufactured by Thermo Fisher Scientific

Sourced in Belgium

Goat anti-rabbit IgG Alexa Fluor Plus 647 is a secondary antibody conjugated with Alexa Fluor Plus 647 dye. It is designed to detect and bind to rabbit primary antibodies, enabling fluorescent detection and visualization in various immunoassays and imaging applications.

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Close spelling variants of this product

Alexa Fluor 647 (1520 citations) Alexa Fluor 647 goat anti-rabbit (127 citations) Alexa Fluor 647 goat anti-rabbit IgG (104 citations) Alexa Fluor Plus 647 (44 citations) Anti-rabbit Alexa Fluor 647 (38 citations) Alexa Fluor 647 anti-rabbit IgG (27 citations) Goat anti-rabbit 647 (24 citations) Goat anti-rabbit IgG-Alexa 647 (22 citations) Alexa Fluor goat anti-rabbit IgG (17 citations) Alexa Fluor 647 anti-goat IgG (11 citations)

6 protocols using goat anti rabbit igg alexa fluor plus 647

Characterization of Protein Tagging Antibodies

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The following commercially available antibodies were used: mouse anti-HA antibody HA.11 (clone 16B12, Biolegend, Cat# 901503, RRID:AB_2565005), rabbit anti-DYKDDDDK (FLAG) tag antibody (clone D6W5B, Cell Signaling Technology Cat# 14793, RRID:AB_2572291), HRP-coupled rabbit anti-E tag antibody (Bethyl Laboratories Cat# A190-133P, RRID:AB_345222), HRP-coupled MonoRab rabbit anti-camelid VHH antibody (clone 96A3F5, GenScript Cat# A01860-200, RRID:AB_2734123), HRP-coupled mouse anti-HA-Tag (clone 6E2, Cell Signaling Technology Cat# 2999, RRID:AB_1264166), goat anti-mouse IgG Alexa Fluor Plus 647 (Thermo Fisher Scientific Cat# A32728, RRID:AB_2633277), goat anti-rabbit IgG Alexa Fluor Plus 647 (Thermo Fisher Scientific Cat# A32733, RRID:AB_2633282).

Koenig P.A., Das H., Liu H., Kümmerer B.M., Gohr F.N., Jenster L.M., Schiffelers L.D., Tesfamariam Y.M., Uchima M., Wuerth J.D., Gatterdam K., Ruetalo N., Christensen M.H., Fandrey C.I., Normann S., Tödtmann J.M., Pritzl S., Hanke L., Boos J., Yuan M., Zhu X., Schmid-Burgk J.L., Kato H., Schindler M., Wilson I.A., Geyer M., Ludwig K.U., Hällberg B.M., Wu N.C, & Schmidt F.I. (2021). Structure-guided multivalent nanobodies block SARS-CoV-2 infection and suppress mutational escape. Science (New York, N.y.), 371(6530), eabe6230.

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Immunofluorescence Analysis of Mouse Tissues

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Immunofluorescence and analysis was performed on formalin-fixed mouse tissues (n = 5 per treatment) by Reveal Biosciences. Each sample was processed and embedded in its own cassette and paraffin-embedded blocks were sectioned at 4 μm onto positively charged slides. Heat-induced antigen retrieval was performed using Leica Bond Epitope Retrieval Buffer 2 (ethylenediaminetetraacetic acid solution, pH 9.0) for 20 min. Nonspecific background was blocked with Novocastra Protein Block (RE7102-CE ; Leica) for 20 min. Each tissue was stained with primary antibodies against CD3, CD8, granzyme B, F4/80, CD11c, and PDL1 (Table 1) for 60 min. Secondary antibodies, goat anti-rabbit IgG Alexa Fluor Plus 647 (A32733; Thermo Fisher) or goat anti-rat IgG Alexa Fluor 647 (A21247; Thermo Fisher), were applied for 30 min. Slides were mounted with DAPI in Fluorogel II for nuclear visualization. Whole slide images were generated in fluorescence using a Pannoramic SCAN (3D Histech). Positive cells and intensity were quantified using AI technology ImageDx (Reveal Biosciences). All tissue and staining artifacts were digitally excluded from quantification. Data are reported as percent positive cells and as intensity.

Luchtel R.A., Bhagat T., Pradhan K., Jacobs WR J.r., Levine M., Verma A, & Shenoy N. (2020). High-dose ascorbic acid synergizes with anti-PD1 in a lymphoma mouse model. Proceedings of the National Academy of Sciences of the United States of America, 117(3), 1666-1677.

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Antibody Staining for Endocytic Markers

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The primary antibodies used in this study were either made in house or were purchased from Cell Signaling Technologies. Stabilin-2 primary antibody was made by the Dr. Paul Weigel lab and was labeled as mAb30,⁶³ (link) and Stabilin-1 primary antibody (clone 911) was a kind gift from Dr. Marko Salmi (University of Turku, Turku, Finland). Anti-mouse immunoglobulin G (IgG) Fab2 Alexa Flour 488 (Cell Signaling Technology, catalog #4408S) was used for detecting these primary antibodies. Primary antibodies for clathrin (catalog #2410S), caveolin 1 (catalog #3238S), early endosomal antigen (EEA1) (catalog #2411S), Rab7 (catalog #9367S), and LAMP1 (catalog #9091S) were purchased from Cell Signaling Technology. Goat anti-rabbit IgG Alexa Fluor Plus 647 (Invitrogen, #A32733) and goat anti-mouse IgG Alexa Fluor 488 (Cell Signaling Technologies, #4408S) secondary antibody was used for detecting these primary antibodies.

Pandey E, & Harris E.N. (2023). Chloroquine and cytosolic galectins affect endosomal escape of antisense oligonucleotides after Stabilin-mediated endocytosis. Molecular Therapy. Nucleic Acids, 33, 430-443.

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Multiparametric Flow Cytometry Panel for Murine and Human Samples

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Antibodies for FACS of murine samples included CD31 (MEC13.3, BD Bioscience), CD45 (clone 30-F11, BD Bioscience), Ter119 (clone TER-119, BD Bioscience), CD249/BP-1 (clone 6C3, eBioscience), THY1.2 (clone 53-2.1, BD Bioscience), CD200 (clone OX-90, BD Bioscience), CD105 (clone MJ7/18, BioLegend), EMB (clone REA501, Miltenyi Biotec). Antibodies for FACS of human samples included CD31 (clone WM59, BioLegend), CD45 (clone HI30, BioLegend), CD235a (clone GA-R2, BioLegend), THY1-1 (clone 5E10, BD Bioscience), CD200 (clone MRC OX-104, BD Bioscience), CD105 (clone 43A3, BioLegend), EMB (Rabbit monoclonal [EPR11417], Abcam), CD146 (clone P1H12, BD Bioscience), Podoplanin (clone LpMab-17, BD Bioscience), CD73 (clone AD2, BD Bioscience), CD164 (clone N6B6, BD Bioscience), HLA-ABC (clone W6/32, BioLegend), B2M(beta2-microglobulin) (Clone 2M2, BioLegend), Goat anti-Rabbit IgG Alexa Fluor Plus 647 (polyclonal, A32733, Invitrogen) and Donkey anti-Rabbit IgG Alexa Fluor Plus 555(polyclonal,A32794, Invitrogen).

Sun J., Hu L., Bok S., Yallowitz A.R., Cung M., McCormick J., Zheng L.J., Debnath S., Niu Y., Tan A.Y., Lalani S., Morse K.W., Shinn D., Pajak A., Li Z., Li N., Xu R., Iyer S, & Greenblatt M.B. (2023). Discovery of a Vertebral Skeletal Stem Cell Driving Spinal Metastases. Research Square.

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Immunohistochemical Labeling of ASO in Fixed Brains

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Fixed brains were frozen in O.C.T. solution (Tissue-Tek) and sectioned at 40 μm using a cryostat. Free floating sections were blocked and then incubated with a previously described pan-ASO antibody that recognizes the ASO backbone [28 (link)] at 1:2000 dilution overnight. After washing, sections were incubated with Alexa Fluor Plus 647 goat anti-rabbit IgG (Invitrogen, A32733, 1:300 dilution) for 1 h, followed by incubation with DAPI (Invitrogen, D1306). After washing, sections were mounted onto slides with Fluoromount-G mounting media (Invitrogen, 00-4958-02). Images were acquired on an Olympus FV1000 confocal microscope with a 20x objective.

Smith D.M., Niehoff M.L., Ling K., Jafar-Nejad P., Rigo F., Farr S.A., Wilkinson M.F, & Nguyen A.D. (2023). Targeting nonsense-mediated RNA decay does not increase progranulin levels in the Grn R493X mouse model of frontotemporal dementia. PLOS ONE, 18(3), e0282822.

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Immunofluorescence Staining of Piglet Liver

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Liver segments of piglets were subjected to immunofluorescence staining. Briefly, six micron-thick slices were obtained from paraffin-embedded livers. Xylene and a series of alcohols were used for section dewaxing. After being soaked in citrate antigen retrieval solution (Beyotime Biotechnology, P0081, Shanghai, China), 0.3% Triton X-100 (Beyotime Biotechnology, ST797), and 3% H2O2-methanol, slices were blocked with 5% BSA for 1–2 h at room temperature. Next, specimens were incubated at 4 °C overnight with rabbit-anti-F4/80 (Servicebio, GB113373, Gent, Belgium) and cultured for 1 h with the secondary antibody, Alexa Fluor Plus 647 goat anti-rabbit IgG (Invitrogen, A32733). After all antigens had been labeled, nuclei were stained with Hoechst 33342 (Invitrogen, H3570). Images were acquired using a confocal laser scanning microscope (Carl Zeiss, Oberkochen, Germany) and analyzed by ImageJ program V1.8.0.112 (National Institutes of Health, Bethesda, MD, USA).

Xu M., Zhuo R., Tao S., Liang Y., Liu C., Liu Q., Wang T, & Zhong X. (2022). M6A RNA Methylation Mediates NOD1/NF-kB Signaling Activation in the Liver of Piglets Challenged with Lipopolysaccharide. Antioxidants, 11(10), 1954.

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