Goat anti rabbit igg secondary antibody

Manufactured by LI COR

Sourced in United States

The Goat anti-rabbit IgG secondary antibody is a laboratory reagent designed to detect the presence of rabbit primary antibodies in various immunological assays. It binds to the Fc region of rabbit immunoglobulin G (IgG) molecules, allowing for the indirect detection and visualization of target antigens.

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Lab products found in correlation

Anti smc1, by Fortis Life Sciences (2 mentions) Goat anti mouse igg secondary antibody, by LI COR (2 mentions) Anti actin, by Thermo Fisher Scientific (2 mentions) Anti histone h3 ps10, by Cell Signaling Technology (1 mentions) Irdye 680rd goat anti mouse igg secondary antibody, by LI COR (1 mentions) Affi prep protein a beads, by Bio-Rad (1 mentions) Anti myc, by Merck Group (1 mentions) Pakt s473, by Cell Signaling Technology (1 mentions) Anti cleaved caspase 1, by Affinity Biosciences (1 mentions) Odyssey clx imaging system, by LI COR (1 mentions) Anti caspase 1, by Affinity Biosciences (1 mentions) Anti il 1β, by Affinity Biosciences (1 mentions) Odyssey version 3, by LI COR (1 mentions) Bca assay kit, by Bio-Rad (1 mentions) Odyssey infrared imaging system, by LI COR (1 mentions) Ripa lysis buffer, by Beyotime (1 mentions) Anti myc, by Roche (1 mentions) Anti cenp a, by Merck Group (1 mentions) Anti h3k4me2, by Merck Group (1 mentions) Anti h3k9me3, by Abcam (1 mentions)

Close spelling variants of this product

Goat anti-rabbit secondary antibody (24 citations) Anti-rabbit secondary antibody (13 citations)

8 protocols using goat anti rabbit igg secondary antibody

Antibody-Based Protein Analysis Protocol

Cited 2 times

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The following antibodies were used in this study: anti-centromere antibody (ACA or CREST-ImmunoVision, HCT-0100), anti-PP2A-Aα (Santa Cruz, Sc-6112), anti-Histone H3-pS10 (Cell signaling, 9706), anti-Smc1 (Bethyl, A300-055A), anti-SET (Bethyl, A302-261A), AKT (Cell signaling, 4691S), pAKT (S473) (Cell signaling, 4060S), anti-actin (Thermo Scientific, MA5-11869), anti-pHec1 (phospho Ser55, GTX70017, GeneTex), and anti-Myc (Millipore, 11667149001). Anti-Sororin is a gift from Susannah Rankin. Anti-Sgo1 and anti-GFP antibodies were made in-house as described previously (Liu et al., 2013b (link); Kim and Yu, 2015 (link)).
Antibody dilution for immunoblotting was often 1:1000 unless specified.
The secondary antibodies were purchased from Li-COR: IRDye 680RD goat anti-mouse IgG secondary antibody (926-68070) and goat anti-rabbit IgG secondary antibody (926-32211).
Harvested cells were collected and lysed with SDS sample buffer. After being 5-min boiled, lysates were resolved by SDS–PAGE and blotted with indicated antibodies.
For immunoprecipitation, anti-Myc or anti-GFP antibodies were coupled to Affi-Prep Protein A beads (Bio-Rad) at a concentration of 1 mg/ml⁻¹.

Yang L., Zhang Q., Niu T, & Liu H. (2021). SET levels contribute to cohesion fatigue. Molecular Biology of the Cell, 32(13), 1256-1266.

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Western Blot Analysis of NLRP3 Inflammasome

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Cell and tissue proteins were extracted as previously described (19 (link)), and bicinchoninic acid method was used to determine the protein concentration. Nitrocellulose membranes were incubated with primary antibodies (anti-NLRP3, anti-IL-18, GAPDH, Proteintech Group, Inc., Chicago, IL,USA; anti-GSDMD, Abbexa Ltd, Cambridge, United Kingdom; anti-caspase-1, anti-IL-1β, anti-cleaved caspase-1, anti-cleaved IL-1β, Affinity Biosciences, Cincinnati, OH, USA; anti-caspase-11 p20, Santa Cruz Biotechnology, Inc.Dallas, Texas, USA) at 4 °C overnight. The membranes were washed with 1% TBST before and after incubation with goat anti-rabbit IgG secondary antibody (LI-COR Biotechnology, Lincoln, NE, USA) or goat anti-mouse IgG secondary antibody (LI-COR Biotechnology, Lincoln, NE, USA) for 1 h at room temperature. Odyssey CLx imaging system (LI-COR Biosciences, Lincoln, NE, USA) was used to analysis protein expression as previously described (19 (link)).

Liu W., Yang D., Shi J., Wen P., Zhang J., Wang Z., Hu B., Shi X., Cao S., Guo W, & Zhang S. (2021). Caspase-1 Inhibitor Reduces Pyroptosis Induced by Brain Death in Kidney. Frontiers in Surgery, 8, 760989.

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Western Blot Analysis of A375 Cell Proteins

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Protein samples from cultured A375 cells were harvested with RIPA lysis buffer (Beyotime Institute of Biotechnology) containing 1% protein inhibitor and 10% phosphatase inhibitor. After being centrifuged at 12,000 × g at 4°C for 15 min, the supernatant was transferred and protein concentrations were assessed using a BCA assay kit (Bio-Rad Laboratories, Inc.). Equal amounts of protein (80 µg/lane) were loaded onto a 8–15% gel, resolved using SDS-PAGE and blotted onto a nitrocellulose membrane. After blocking with 5% non-fat milk at 4°C for 2 h, the membranes were incubated with primary antibodies at 4°C overnight. The primary antibodies were anti-p-STAT3 (Y705, #9131, 1:1,000), anti-STAT3 (#4904, 1:1,000), anti-p-AMPK (Thr172, #2535, 1:1,000), anti-AMPK (#2532, 1:1,000), anti-p62 (#88588, 1:1,000), anti-Bcl-2 (ab196495, 1:1,000) and anti-LC3 (L7543, 1:1,000). After being rinsed with TBS-0.1% Tween 20 (5 min for 3 times), the membranes were subsequently incubated with fluorescence-conjugated goat anti-mouse IgG or goat anti-rabbit IgG secondary antibody (1:10,000;LI-COR Biosciences) for 1 h at room temperature. Images were captured using an Odyssey infrared imaging system and Odyssey version 3.0 software (LI-COR Biosciences).

Jin J., Chen N., Pan H., Xie W., Xu H., Lei S., Guo Z., Ding R., He Y, & Gao J. (2020). Triclosan induces ROS-dependent cell death and autophagy in A375 melanoma cells. Oncology Letters, 20(4), 73.

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Antibody Characterization Protocol

Cited 2 times

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The following antibodies were used in this study: anti-centromere antibody (ACA; or CREST-ImmunoVision; HCT-0100), anti-Myc (Roche; 11667203001), anti-Smc1 (Bethyl; A300-055A), anti-Rpb1 (Abcam; ab5408), anti-H3K4me2 (EMD Millipore; 07–030), anti-H3K9me3 (Abcam; ab8898), anti-Actin (Invitrogen; MA5-11869), anti-CENP-B (Abcam; ab25734), anti–CENP-A (EMD Millipore; 07–574), and anti–Rpb2-pSer2 (BioLegend; H5). Anti-Sgo1, anti-Bub1, and anti-Wapl were made in-house as described previously (Liu et al., 2013a (link); Qu et al., 2019 (link)). Anti-Sororin antibodies described previously were a gift from Dr. Susannah Rankin at Oklahoma Medical Research Foundation, Oklahoma City, OK (Liu et al., 2013b (link)).
The secondary antibodies were purchased from LI-COR: Goat anti-Mouse IgG Secondary Antibody (926–68070) and Goat anti-Rabbit IgG Secondary Antibody (926–32211).
For immunoblotting, primary and secondary antibodies were used at 1-µg ml⁻¹ concentration.

Chen Y., Zhang Q., Teng Z, & Liu H. (2021). Centromeric transcription maintains centromeric cohesion in human cells. The Journal of Cell Biology, 220(7), e202008146.

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Western Blot Optimization and Visualization

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SDS-PAGE separation of proteins was performed using 12% or 15% (v/v) acrylamide gels following standard procedures [78 (link)]. After separation of ~10 μg protein by SDS-PAGE, proteins were transferred to a nitrocellulose membrane by electro-blotting using 10 mM CAPS [3-(cyclohexylamino)-1-propanesulfonic acid; pH 11], 10% methanol transfer buffer. Membranes were blocked for 1 hr at RT with 50% (v/v) Odyssey blocking buffer in PBS (Licor, USA). Optimal conditions for each rabbit polyclonal antibody were determined empirically and the reproducibility of the results verified by repeated Western blot analyses during these optimisations (data not shown). Membranes were incubated with primary rabbit polyclonal antibodies, in a range from 1:500–1:15000, in blocking buffer containing 0.12–0.25% (v/v) Tween-20 for 1 hr at RT or overnight at 4°C. Following 4 × 5 min washes in 0.1% (v/v) Tween-20, the membranes were incubated in 1:7500 fluorescently labelled goat anti-rabbit IgG secondary antibody (Licor, USA) in blocking buffer containing 0.12–0.25% Tween-20. Membranes were washed again and visualised on a Licor Odyssey imaging system (USA).

Stockdale S.R., Collins B., Spinelli S., Douillard F.P., Mahony J., Cambillau C, & van Sinderen D. (2015). Structure and Assembly of TP901-1 Virion Unveiled by Mutagenesis. PLoS ONE, 10(7), e0131676.

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Western Blot Analysis of Apoptosis and Oxidative Stress Markers

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The total proteins were separated on SDS-PAGE gel, blotted onto PVDF membrane, and then blocked with 5% nonfat milk. Subsequently, the membranes were incubated with antibodies Bax (1 : 1000, Abcam), Bcl-2 (1 : 1000, Abcam), Nrf2 (1 : 1000, Abcam), HO-1 (1 : 1000, Abcam), and β-actin (1 : 1000, Abcam). The membranes were incubated goat anti-rabbit IgG secondary antibody (LI-COR) and analyzed by a two-color infrared imaging system (Odyssey; LICOR) to quantify protein expression.

Jiang L., Gong Y., Rao J., Yang Q., Gao N., Li G, & Ma Y. (2021). 1-O-Hexyl-2,3,5-Trimethylhydroquinone Ameliorates the Development of Preeclampsia through Suppression of Oxidative Stress and Endothelial Cell Apoptosis. Oxidative Medicine and Cellular Longevity, 2021, 8839394.

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PTEN Redox State Analysis

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Whole-cell lysates were prepared from 5x10 6 SeptMBr cells treated with Nacetylcysteine (MilliporeSigma A7250) for 4 hours followed by stimulation/oxidation with 1 mM hydrogen peroxide (Fisher Scientific Canada, Mississauga ON) for 5 minutes. Cells were washed and resuspended in PBS with 10% trichloroacetic acid (MillipreSigma T9159), sonicated, and washed again in 100% acetone. To preserve PTEN protein in its reduced state, protein precipitates were resolubilized in non-reducing lysis buffer (2% SDS, 50 mM Tris-HCl pH 6.8, 10% glycerol, 0.1% bromophenol blue) containing 40 mM N-ethylmaleimide (MilliporeSigma E3876). Samples were denatured at 100°C and separated on 10% SDS-PAGE gels with BLUeye Prestained Protein Ladder (FroggaBio, Concord ON). Protein was then semidry transferred to a PVDF membrane and incubated in Li-Cor Intercept ® Blocking Buffer (Cedarlane, Burlington ON). Immunoblotting was performed with monoclonal anti-PTEN rabbit IgG primary antibody (Cell Signalling Technology 9188S, Danvers MA) at 4°C overnight and goat anti-rabbit IgG secondary antibody (Li-Cor 926-32211) for 1 hour at room temperature.
Blots were visualized with the Odyssey CLx Infrared imaging system at 800 nm (Li-Cor).

Sams M.P., Iansavitchous J., Astridge M., Rysan H., Xu L.S., Rodrigues de Oliveira B, & DeKoter R.P. (2024). N-Acetylcysteine Alters Disease Progression and Increases Janus Kinase Mutation Frequency in a Mouse Model of Precursor B-Cell Acute Lymphoblastic Leukemia. The Journal of pharmacology and experimental therapeutics, 389(1).

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Quantifying GABA Receptor Proteins in Mouse Hippocampus

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Total proteins were extracted from the hippocampus of mice using the Protein Extraction Kit (cat. no. KGP2100; Nanjing KeyGen Biotech Co., Ltd.) and the protein concentration was measured using the BCA Protein Assay Kit (cat. no. KGP902; Nanjing KeyGen Biotech Co., Ltd.). Equal amounts of protein (60–80 µg/lane) were separated by SDS-PAGE on 8 or 10% gels and were then transferred to PVDF membranes. When the protein transfer was completed, membranes were blocked with 5% non-fat milk for 1 h in room temperature, followed by incubation for ~20 h at 4°C with rabbit anti-GABABR2 (1:500; cat. no. ab230136), anti-glutamic acid decarboxylase (GAD)65/67 (1:500 cat. no. ab183999) and anti-GAPDH (1:1,000; cat. no. ab181602) antibodies. Subsequently, the membranes were washed with TBS-Tween (0.5%) three times (5 min/wash) and were further incubated with the corresponding goat anti-rabbit IgG secondary antibody (1:1,000; LI-COR Biosciences cat. no. P/N: 926-32211) for 2 h in room temperature. GAPDH served as an internal reference. Semi-quantification of bands was performed from optical density values using the Odyssey CLX instrument system (LI-COR Biosciences).

Jiang S., Xiao L., Sun Y., He M., Gao C., Zhu C., Chang H., Ding J., Li W., Wang Y., Sun T, & Wang F. (2022). The GABAB receptor agonist STX209 reverses the autism-like behaviour in an animal model of autism induced by prenatal exposure to valproic acid. Molecular Medicine Reports, 25(5), 154.

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