Irdye 680rd goat anti rabbit secondary antibody

Manufactured by LI COR

Sourced in United States, Germany

The IRDye 680RD goat anti-rabbit secondary antibody is a fluorescently labeled detection reagent. It is designed to bind to and detect the presence of rabbit primary antibodies in various immunoassays and imaging applications.

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Lab products found in correlation

Propidium iodide, by Thermo Fisher Scientific (3 mentions) Donkey anti mouse 800cw secondary antibody, by LI COR (3 mentions) Infrarot imaging system, by LI COR (2 mentions) Image studio lite, by LI COR (2 mentions) Penicillin streptomycin, by Thermo Fisher Scientific (2 mentions) Dulbecco s modified eagle medium, by Thermo Fisher Scientific (2 mentions) Odyssey clx image reader, by LI COR (2 mentions) Pvdf membrane, by Bio-Rad (2 mentions) Rabbit monoclonal primary antibody against glut4, by Abcam (2 mentions) Gapdh, by Cell Signaling Technology (2 mentions) Hoechst 33342, by Thermo Fisher Scientific (2 mentions) Irdye 680rd goat anti mouse secondary antibody, by LI COR (1 mentions) Horseradish peroxidase coupled anti rabbit or anti mouse antibodies, by Merck Group (1 mentions) Irdye 800cw goat anti mouse igg, by LI COR (1 mentions) Nitrocellulose membrane, by Thermo Fisher Scientific (1 mentions) Rabbit anti β actin, by Abcam (1 mentions) Mouse anti e cadherin, by Abcam (1 mentions) Salinomycin, by Merck Group (1 mentions) Triton x 100, by Thermo Fisher Scientific (1 mentions) Pageruler prestained protein ladder, by Thermo Fisher Scientific (1 mentions)

Close spelling variants of this product

IRDye secondary antibodies (362 citations) IRDye 680RD (357 citations) IRDye 680RD goat anti-rabbit (108 citations) Goat anti-rabbit secondary antibody (24 citations) IRDye 680RD secondary antibody (20 citations) Anti-rabbit IRDye 680RD (17 citations) IRDye 680RD goat (17 citations) Anti-rabbit secondary antibody (13 citations) IRDye 680RD goat anti-rabbit IgG secondary antibody (13 citations) IRDye goat anti-rabbit (11 citations)

12 protocols using irdye 680rd goat anti rabbit secondary antibody

Immunoblotting of Apoptosis Regulators

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PVDF membranes were incubated with (1:500 dilution) Anti-BAD Antibody
conjugated to Alexa-Fluor 680 (C-7) (Santa Cruz Biotechnology, sc-8044
AF680), (1:500 dilution) Anti-TP53BP2 Antibody conjugated to Alexa-Fluor
680 (Santa Cruz Biotechnology, sc-398311 AF680), (1:500 dilution)
Anti-pan14-3-3 Antibody (Santa Cruz Biotechnology, sc-1657), and (1:1000
dilution) Anti-Cereblon Antibody (Cell Signaling Technology, D8H3S)
overnight at 4 °C. Alexa-Fluor conjugated antibodies were directly
detected on the blots, after overnight incubation following washing
of the membrane. Blots treated with either Anti-pan14-3-3 Antibody
or Anti-Cereblon Antibody were washed after overnight incubation and
then incubated with IRDYE 680RD Goat Anti-Mouse Secondary Antibody
(LI-COR, 926-68070) or IRDYE 680RD Goat Anti-Rabbit Secondary Antibody
(LI-COR, 926-68071).

Zhu P., Stanisheuski S., Franklin R., Vogel A., Vesely C.H., Reardon P., Sluchanko N.N., Beckman J.S., Karplus P.A., Mehl R.A, & Cooley R.B. (2023). Autonomous Synthesis of Functional, Permanently Phosphorylated Proteins for Defining the Interactome of Monomeric 14-3-3ζ. ACS Central Science, 9(4), 816-835.

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Immunoblot Analysis of Peroxisomal Proteins

Cited 2 times

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Immunoblot analyses were performed according to standard procedures. Polyclonal rabbit antibodies used were raised against Pex5p (Albertini et al. 1997 (link)), Pex14p (Girzalsky et al. 1999 (link)), Pex13p (Girzalsky et al. 1999 (link)) and Pex17p (Huhse et al. 1998 (link)). To detect GFP, monoclonal mouse antibodies (Sigma-Aldrich/Merck or Roche Diagnostics, Germany) were used. Immuno-reactive complexes were visualized either using the IRDye 800CW goat anti-mouse IgG or IRDye 680RD goat anti-rabbit secondary antibody (Li-COR Biosciences, Bad Homburg, Germany) followed by detection with an ‘Infrarot Imaging System’ (Li-COR Biosciences) or using horseradish-peroxidase coupled anti-rabbit or anti-mouse antibodies (Sigma-Aldrich/Merck, Germany) and subsequent detection of chemiluminescence signals with a ChemoCam Camera system (INTAS Science instruments GmbH, Göttingen, Germany). Quantification of immunoblot signals (Supplementary Figure 1E; n = 3) was performed using the software Image Studio Lite (LI-COR Biosciences).

Fischer S., Bürgi J., Gabay-Maskit S., Maier R., Mastalski T., Yifrach E., Obarska-Kosinska A., Rudowitz M., Erdmann R., Platta H.W., Wilmanns M., Schuldiner M., Zalckvar E., Oeljeklaus S., Drepper F, & Warscheid B. (2022). Phosphorylation of the receptor protein Pex5p modulates import of proteins into peroxisomes. Biological Chemistry, 404(2-3), 135-155.

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Curcumin and Salinomycin Cancer Cell Assay

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Dulbecco's Modified Eagle Medium (DMEM), fetal bovine serum (FBS), penicillin–streptomycin, propidium iodide, Triton X-100, PageRuler Prestained Protein Ladder, and nitrocellulose membrane were purchased from Thermo Fisher. Curcumin, salinomycin, 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDC), and N-hydroxysuccinimide (NHS) were bought from Sigma-Aldrich and used directly without any purification. Mouse anti-vimentin, mouse anti-E-cadherin, and rabbit anti-β-actin primary antibodies purchased from Abcam. Odyssey blocking buffer, IRDye® 800CW goat anti-mouse secondary antibody, and IRDye® 680RD goat anti-rabbit secondary antibody were purchased from LI-COR.

Zhao Y., Wang K., Zheng Y., Zeng X., Lim Y.C, & Liu T. (2021). Co-delivery of Salinomycin and Curcumin for Cancer Stem Cell Treatment by Inhibition of Cell Proliferation, Cell Cycle Arrest, and Epithelial–Mesenchymal Transition. Frontiers in Chemistry, 8, 601649.

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Astrocyte Protein Expression Analysis

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ONH astrocytes in 6-well plates were treated, washed with ice-cold 1X DPBS (Gibco, Thermo Fisher Scientific), and lysates were scraped into 2X Laemmli sample buffer (90% sample buffer, 10% β-mercaptoethanol (BME; Fisher Chemical, 034461-100)). Samples were boiled for 5 min, loaded in equal amounts (10 μg) into NuPAGETM 4–12% Bis-Tris Gels (Invitrogen; NP0321), and proteins separated using SDS-PAGE at a constant rate of 180 V for 80 (+30) min. Proteins in gel slabs were transferred electrophoretically to 0.45 μm nitrocellulose Immobilon-FL membranes (Sigma; IPFL00010). Membranes were blocked with 5% bovine serum albumin (Thermo Fisher Scientific; BP9706) in trisbuffered saline with 0.2% Tween®20 (Thermo Fisher Scientific), and probed with primary antibodies against GFAP (rabbit polyclonal GFAP antibody, 1:1000; Novus Biologicals, NB300-141) and fibronectin (rabbit anti-fibronectin antibody, 1:50000, Abcam, Ab45688) followed by incubation with an IRDye 680RD goat-anti-rabbit secondary antibody (1:15000, LI-COR, 926–68071). Bound antibodies were visualized using the Li-Cor Odyssey CLx Infrared imaging system. Densitometry of target antibody bands was performed using ImageStudioLite software and normalized to correspondent GAPDH bands (anti-GAPDH [G9545] 1:80000; Sigma-Aldrich).

Kirschner A., Strat A.N., Yablonski J., Yoo H., Bagué T., Li H., Zhao J., Bollinger K.E., Herberg S, & Ganapathy P.S. (2021). Mechanosensitive channel inhibition attenuates TGFβ2-induced actin cytoskeletal remodeling and reactivity in mouse optic nerve head astrocytes. Experimental eye research, 212, 108791.

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Immunoblot Analysis of Histone Modifications

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Immunoblotting was performed as previously described (Honda and Selker, 2008 (link)). Briefly, Neurospora extracts were produced by sonication in extraction buffer (50 mM Hepes pH7.5, 1 mM EDTA, 150 mM NaCl, 10% Glycerol, 0.02% NP40) supplemented with cOmplete ULTRA protease inhibitor cocktail tablets (Roche, 05892970001). The following antibodies were used for immunoblotting: H3K36me3 (Cell Signaling, Cat#4909S, Clone (D5A7)), H3K36me2 (Abcam, Cat#ab9049), Histone H3 (Abcam, Cat#ab1791), IRDye 680RD Goat-anti-Rabbit secondary antibody (Licor, Cat#926 – 68071).

Bicocca V.T., Ormsby T., Adhvaryu K.K., Honda S, & Selker E.U. (2018). ASH1-catalyzed H3K36 methylation drives gene repression and marks H3K27me2/3-competent chromatin. eLife, 7, e41497.

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Immunoblotting Analysis of Peroxisomal Proteins

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Polyclonal rabbit antibodies were raised against Pex13 [31 (link)] and Por1 [32 (link)]. Monoclonal mouse antibodies were raised against GFP (Sigma-Aldrich/Merck, Germany) and Pgk1 (Invitrogen, Karlsruhe, Germany). Immuno-reactive complexes were visualized using the IRDye 800CW goat anti-rabbit IgG or IRDye 680RD goat anti-rabbit secondary antibody (Li-COR Bioscience, Bad Homburg, Germany) followed by detection using the “Infrarot Imaging System“ (Li-COR Bioscience, Bad Homburg, Germany). The intensity of free anti-Pex13 signals on the Western blots was calculated by Image Studio Lite, LI-COR Bioscience.

Mastalski T., Brinkmeier R, & Platta H.W. (2020). The Peroxisomal PTS1-Import Defect of PEX1- Deficient Cells Is Independent of Pexophagy in Saccharomyces cerevisiae. International Journal of Molecular Sciences, 21(3), 867.

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Western Blot Protein Detection Protocol

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Cells were lysed in RIPA lysis buffer (50 mM Tris-Cl pH 8.0, 150 mM NaCl, NP-40, 0.5% sodium deoxycholate, 0.1% SDS) and 1× protease inhibitor cocktail (Roche, Indianapolis, IN, USA). Protein concentrations were quantified with the Bradford protein assay. Briefly, 25 μg of protein extracts were loaded and separated by SDS-PAGE gels. Blotting was performed with standard protocols using a PVDF membrane (Bio-Rad, Hercules, CA, USA). Membranes were blocked for 1 h in blocking buffer (5% BSA in PBST) and probed with primary antibodies at 1:1,000 dilution at 4 °C overnight. After three washes with PBST, the membranes were incubated with diluted goat anti-rabbit secondary IRDye 680RD antibody at 1:10,000 (LI-COR, Lincoln, NE, USA) for 1 h at room temperature. After washing, membranes were visualized on LI-COR Odyssey CLx image reader. All antibodies used for immunoblotting are listed in the supplementary section.

Moreno R.Y., Juetten K.J., Panina S.B., Butalewicz J.P., Floyd B.M., Venkat Ramani M.K., Marcotte E.M., Brodbelt J.S, & Zhang Y.J. (2023). Distinctive interactomes of RNA polymerase II phosphorylation during different stages of transcription. iScience, 26(9), 107581.

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Western Blot Analysis of Protein Expression

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Briefly, cells were lysed in RIPA buffer (10 mM Tris–HCl, pH 7.5, 150 mM NaCl, 1% sodium deoxycholate, 0.1% SDS, 1% Triton x-100, 5 mM EDTA) supplemented with 1 × Halt protease and phosphatase inhibitor cocktail (ThermoFisher) followed by centrifugation for 15 min at 12,000 rpm. Protein concentration in supernatants was measured with BCA assay. Typically, proteins (50 µg) were separated by Novex 4–12% Tris–Glycine gels (Invitrogen, Waltham, MA), transferred to PVDF membranes (Bio-Rad), followed by membrane blocking at room temperature for 1 h in 1% Tween-20-TBS buffer containing 5% BSA (bovine serum albumin, ThermoFisher) or non-fat milk. Membranes were incubated at 4 °C overnight with primary rabbit antibodies against REST (#22242–1-AP, Proteintech, Rosemont, IL) at 1:500 dilution, SCP1 (#ab136038, Abcam, Cambridge, MA) at 1:500 dilution, or β-tubulin (#ab6046, Abcam) at 1:4000 dilution. On the next day, membranes were washed and incubated with 1:15,000 diluted goat anti-rabbit secondary IRDye 680RD antibody (LI-COR, Lincoln, NE) for 1 h at room temperature. After washing, membranes were visualized on LI-COR Odyssey CLx image reader.

Panina S.B., Schweer J.V., Zhang Q., Raina G., Hardtke H.A., Kim S., Yang W., Siegel D, & Zhang Y.J. (2024). Targeting of REST with rationally-designed small molecule compounds exhibits synergetic therapeutic potential in human glioblastoma cells. BMC Biology, 22, 83.

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Antibody Characterization for Signaling Proteins

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Rabbit monoclonal primary antibodies against AT1R, phospho‐insulin receptor 1 (Tyr632, Ser307), insulin receptor 1, p67 phox, and p47phox were obtained from Santa Cruz Biotechnology Inc (SantaCruz, CA, USA). Rabbit monoclonal primary antibodies against AT2R, rabbit monoclonal primary antibody against GLUT4, and mouse monoclonal primary antibody against α‐tubulin were bought from Abcam (Cambridge, MA, USA). Rabbit monoclonal primary antibody against phospho‐Akt (Ser473), Akt was bought from Cell Signaling Technology Inc. (Danvers, MA, USA). Goat anti‐rabbit IRDye 680RD secondary antibody or Donkey anti‐mouse 800CW secondary antibody was purchased from Licor Biosciences (Lincoln, NE, USA).

Son M., Chan C.B, & Wu J. (2018). Egg White Ovotransferrin‐Derived ACE Inhibitory Peptide Ameliorates Angiotensin II‐Stimulated Insulin Resistance in Skeletal Muscle Cells. Molecular Nutrition & Food Research, 62(4), 1700602.

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Tripeptide IRW Synthesis and Evaluation

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Tripeptide IRW was synthesized with 99.9% purity validated by HPLC-MS/MS by Genscript (Piscataway, NJ, USA). Angiotensin II, dithiothreitol (DTT), Triton X-100, and alkaline phosphatase activity fluorometric assay kit were purchased from Sigma-Aldrich (St. Louis, MO, USA). Cell growth media α-MEM (A10490), fetal bovine serum (FBS), and Pen-Strep solution were purchased from Gibco/Invitrogen (Carlsbad, CA, USA). Dihydrethidium (DHE) and Hoechst 33342 were purchased from Thermo Fisher Scientific (Thermo Fisher Scientific, Burlington, ON, Canada). Annexin V-FITC apoptosis staining kit was purchased from Abcam (Cambridge, MA, USA). Rabbit monoclonal primary antibodies against AT1R, AT2R, MasR, OPG, NFκB, and ALP were obtained from Santa Cruz Biotechnology Inc. (Santa Cruz, CA, USA). Rabbit monoclonal primary antibodies against COL1A2, COX2, AT2R, RUNX2, ACE, and ACE2 were bought from Abcam (Cambridge, MA, USA). The RANKL and GAPDH were purchased from Cell signaling technology (Danvers, MA, USA). Goat anti-rabbit IRDye 680RD secondary antibody and Donkey anti-mouse 800CW secondary antibody was purchased from Licor Biosciences (Lincoln, NE, USA). All the remaining supplies used in this study were obtained from Sigma-Aldrich (St. Louis, MO, USA).

Shang N., Bhullar K.S, & Wu J. (2022). Tripeptide IRW Protects MC3T3-E1 Cells against Ang II Stress in an AT2R Dependent Manner. Molecules, 27(12), 3684.

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