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Goat α rabbit igg hrp

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Goat α-rabbit IgG-HRP is a secondary antibody conjugated with horseradish peroxidase (HRP). It is used to detect and quantify the presence of rabbit immunoglobulin G (IgG) in various immunoassay applications.

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Lab products found in correlation

Mouse α flag m2 antibody ab, by Merck Group (3 mentions) Affinity gel, by Merck Group (3 mentions) α fyn, by Cell Signaling Technology (3 mentions) α ptyr416 src, by Cell Signaling Technology (3 mentions) α ptyr412 abl, by Cell Signaling Technology (3 mentions) Goat α mouse igg hrp, by Merck Group (3 mentions) α flag, by Cell Signaling Technology (3 mentions) Horseradish peroxidase hrp conjugated secondary antibody, by Merck Group (2 mentions) α src, by Cell Signaling Technology (2 mentions) α alpha tubulin, by Cell Signaling Technology (2 mentions) α pan actin, by Cell Signaling Technology (2 mentions) Light chain specific goat α mouse igg hrp, by Merck Group (2 mentions) α phosphotyrosine 4g10, by Merck Group (2 mentions) α myc 9e10, by American Type Culture Collection (2 mentions) α α tubulin, by Cell Signaling Technology (1 mentions) Rabbit αgst, by Merck Group (1 mentions) Mini gel tank system, by Thermo Fisher Scientific (1 mentions) Mini blot module, by Thermo Fisher Scientific (1 mentions) Western lightning plus ecl enhanced chemiluminescence substrate, by PerkinElmer (1 mentions) Bolt 4 12 bis tris plus gel, by Thermo Fisher Scientific (1 mentions)

5 protocols using goat α rabbit igg hrp

1

Antibody-based Western Blotting Assay

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The mouse α-Flag (M2) antibody (Ab) and Affinity Gel were from Sigma and the free Ab was used for Western blotting at a concentration of 0.5 μg/mL. Cell Signaling Technologies Inc. (Danvers, MA, USA) was the source of the following antibodies, used at the indicated dilution or concentration: α-Flag (M2, rabbit mAb; 1:1000), α-Fyn (rabbit mAb; 1:2000), α-Src (rabbit mAb; 1:2000), α-pTyr416-Src (rabbit mAb; 1:5000), α-pTyr412-Abl (rabbit mAb; 1:1000), α-alpha-tubulin (1:1000), and α-pan-actin (0.1 μg/mL). The rabbit α-Abl antibody (K-12, 0.2 μg/mL) was from Santa Cruz Biotechnology (Dallas, TX, USA). The α-phosphotyrosine (4G10; 1:1000) was from EMD Millipore (Billerica, MA, USA) and the α-Myc (9E10; 1:1000) was obtained from American Type Tissue Collection (Manassas, VA, USA). All primary antibodies were diluted in 1.5% BSA in Tris-Buffered Saline (0.9% NaCl, 0.4% Tris-HCl, and 0.1% Tris-base) with 0.05% Tween 20 (TBS-T) and containing 0.005% sodium azide. Horseradish peroxidase (HRP)-conjugated secondary antibodies were obtained from EMD Millipore and used at the following concentrations: goat α-mouse IgG-HRP (1:5,000), light-chain-specific goat α-mouse IgG-HRP (1:10,000) and goat α-rabbit IgG-HRP (1:15,000). All secondary antibodies were diluted in TBS-T.
Schmoker A.M., Weinert J.L., Kellett K.J., Johnson H.E., Joy R.M., Weir M.E., Ebert A.M, & Ballif B.A. (2017). Dynamic multi-site phosphorylation by Fyn and Abl drives the interaction between CRKL and the novel scaffolding receptors DCBLD1 and DCBLD2. The Biochemical journal, 474(23), 3963-3984.
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2

Antibody-based Western Blotting Assay

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The mouse α-Flag (M2) antibody (Ab) and Affinity Gel were from Sigma and the free Ab was used for Western blotting at a concentration of 0.5 μg/mL. Cell Signaling Technologies Inc. (Danvers, MA, USA) was the source of the following antibodies, used at the indicated dilution or concentration: α-Flag (M2, rabbit mAb; 1:1000), α-Fyn (rabbit mAb; 1:2000), α-Src (rabbit mAb; 1:2000), α-pTyr416-Src (rabbit mAb; 1:5000), α-pTyr412-Abl (rabbit mAb; 1:1000), α-alpha-tubulin (1:1000), and α-pan-actin (0.1 μg/mL). The rabbit α-Abl antibody (K-12, 0.2 μg/mL) was from Santa Cruz Biotechnology (Dallas, TX, USA). The α-phosphotyrosine (4G10; 1:1000) was from EMD Millipore (Billerica, MA, USA) and the α-Myc (9E10; 1:1000) was obtained from American Type Tissue Collection (Manassas, VA, USA). All primary antibodies were diluted in 1.5% BSA in Tris-Buffered Saline (0.9% NaCl, 0.4% Tris-HCl, and 0.1% Tris-base) with 0.05% Tween 20 (TBS-T) and containing 0.005% sodium azide. Horseradish peroxidase (HRP)-conjugated secondary antibodies were obtained from EMD Millipore and used at the following concentrations: goat α-mouse IgG-HRP (1:5,000), light-chain-specific goat α-mouse IgG-HRP (1:10,000) and goat α-rabbit IgG-HRP (1:15,000). All secondary antibodies were diluted in TBS-T.
Schmoker A.M., Weinert J.L., Kellett K.J., Johnson H.E., Joy R.M., Weir M.E., Ebert A.M, & Ballif B.A. (2017). Dynamic multi-site phosphorylation by Fyn and Abl drives the interaction between CRKL and the novel scaffolding receptors DCBLD1 and DCBLD2. The Biochemical journal, 474(23), 3963-3984.
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3

Western Blotting Antibody Characterization

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The mouse α-FLAG (M2) antibody (Ab) and Affinity Gel were from Sigma and the free Ab was used for Western blotting at a concentration of 0.5 μg/ml. The rabbit α-GST (1:2000 for Western blotting) was also from Sigma. Cell Signaling Technologies Inc. (Danvers, MA) was the source of the following antibodies, used at 1:1000 dilutions: α-FLAG (M2, rabbit mAb), α-MYC (71D10, rabbit mAb), α-FYN (rabbit polyclonal), α-pTyr416-Src (D49G4, rabbit mAb), α-α-tubulin (DM1A, mouse mAb), α-pTyr412-ABL (247C7, rabbit mAb), α-c-ABL (rabbit polyclonal), α-RXXpS/pT (110B7E, rabbit mAb), and α-RXXpSXP (rabbit polyclonal). The α-pY (4G10; 1:1000) was from EMD Millipore (Billerica, MA). For immunoblotting all primary antibodies were diluted in 1.5% BSA in Tris-Buffered Saline (0.9% NaCl, 0.4% Tris-HCl, and 0.1% Tris-base) with 0.05% Tween 20 (TBS-T) and containing 0.005% sodium azide. Horseradish peroxidase (HRP)-conjugated secondary antibodies were obtained from EMD Millipore and used at the following concentrations: goat α-mouse IgG-HRP (1:5,000), light-chain-specific goat α-mouse and α-rabbit IgG-HRP (1:10,000), and goat α-rabbit IgG-HRP (1:15,000). All secondary antibodies were diluted in TBS-T.
Schmoker A.M., Weinert J.L., Markwood J.M., Albretsen K.S., Lunde M.L., Weir M.E., Ebert A.M., Hinkle K.L, & Ballif B.A. (2020). FYN and ABL Regulate the Interaction Networks of the DCBLD Receptor Family. Molecular & Cellular Proteomics : MCP, 19(10), 1586-1601.
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4

Western Blot Analysis of Bacterial Proteins

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Protein samples were loaded into a Bolt™ 4–12% Bis-Tris Plus Gel (Invitrogen™) with MOPS 1X buffer supplemented with Bolt™ Antioxidant (0.25%), and a Mini Gel Tank system (Invitrogen™) was used. Transference to nitrocellulose membranes was performed using the Mini Blot Module (Invitrogen™). For protein detection, membranes were blocked for 1 h with blocking solution (TBS + 0.1% Tween-20 (TBS-T) + 5% of non-fat powdered milk) and incubated overnight at 4 °C with the following primary antibodies: rabbit α-InlA 1:5000 (Cusabio) or rabbit α-EF-Tu 1:40000 (Abcam) in TBS-T. The secondary antibody goat α-rabbit IgG-HRP 1:20,000 (Sigma) was incubated for 1 h at 4 °C. Detection was performed by chemiluminescence using Western Lightning® Plus-ECL Enhanced Chemiluminescence Substrate (PerkinElmer) in iBright™ CL1500 Imaging System (Supplementary Fig. 5). Images were quantified using Fiji software.
Quendera A.P., Pinto S.N., Pobre V., Antunes W., Bonifácio V.D., Arraiano C.M, & Andrade J.M. (2023). The ribonuclease PNPase is a key regulator of biofilm formation in Listeria monocytogenes and affects invasion of host cells. NPJ Biofilms and Microbiomes, 9, 34.
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5

Protein Expression Analysis in SMA

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Samples, SMA4 patient (proband II:2) and controls (n = 3), were prepared with 4x Laemmli sample buffer (BioRad) and 2-mercaptoethanol (Sigma) solution 9:1 (v/v). Cell lysates (15 μg) were size fractioned using mini-PROTEAN TGX Precast Gels (BioRad) and then transferred to an Immobilon®-P Polyvinylidene difluoride membrane (Sigma). Membranes were probed with the following primary antibodies: rabbit α-CAPN1 (CST), 1:1,500; mouse α-SMN (BD Transduction Laboratories), 1:2000; rabbit α-p-Akt (CST), 1:1,000; mouse α-Pan-Akt (CST), 1:2000; rabbit α-LC3B-II (CST) 1:1,000; rabbit α-β-actin (CST) 1:2000; rabbit α-caspase-3 (Abcam) 1:4,000 and rabbit α-β-actin (CST) 1:2000. Secondary antibodies used in this study include goat α-rabbit IgG HRP (Sigma), 1:10,000 and goat α-mouse IgG HRP (abcam) 1:5,000. Immobilon™ Western Chemiluminescent HRP Substrate Reagent (Millipore) was added and protein bands were visualized using a ChemiDoc™ MP Imaging System (BioRad).
Perez-Siles G., Ellis M., Ashe A., Grosz B., Vucic S., Kiernan M.C., Morris K.A., Reddel S.W, & Kennerson M.L. (2022). A Compound Heterozygous Mutation in Calpain 1 Identifies a New Genetic Cause for Spinal Muscular Atrophy Type 4 (SMA4). Frontiers in Genetics, 12, 801253.
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