Goat anti rat igg h l hrp

Manufactured by Thermo Fisher Scientific

Goat Anti-Rat IgG H + L HRP is a secondary antibody conjugated with horseradish peroxidase (HRP). It is designed to detect and quantify rat immunoglobulins (IgG) in various immunoassays and research applications.

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Lab products found in correlation

Goat anti rabbit igg h l hrp, by Promega (1 mentions) Goat anti mouse igg h l hrp, by Promega (1 mentions) Mouse anti gapdh, by Developmental Studies Hybridoma Bank (1 mentions) Rabbit anti insulin receptor β, by Cell Signaling Technology (1 mentions) Rabbit anti lrp6, by Cell Signaling Technology (1 mentions) Rabbit anti axin1, by Cell Signaling Technology (1 mentions) Rabbit anti flag, by Proteintech (1 mentions) Rat anti ha, by Roche (1 mentions) Mouse anti tubulin, by Developmental Studies Hybridoma Bank (1 mentions) Mouse anti β catenin, by BD (1 mentions) Beyoecl moon, by Beyotime (1 mentions) Polyvinylidene fluoride pvdf membrane, by Thermo Fisher Scientific (1 mentions) Ripa lysis buffer, by Beyotime (1 mentions) Bca protein assay kit, by Beyotime (1 mentions) Goat anti mouse igg h l hrp, by Thermo Fisher Scientific (1 mentions) Multiskan fc microplate reader, by Thermo Fisher Scientific (1 mentions)

Close spelling variants of this product

Goat anti-rat IgG (H + L; Goat anti-rat HRP H + L IgG Anti-rat-HRP

4 protocols using goat anti rat igg h l hrp

Western Blot Analysis of ECM Proteins

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Protein samples were prepared by adding SDS loading buffer with 1 mM DTT for 10 min at 75 °C to whole cell lysate. Proteins were separated by SDS-PAGE prior transfer onto nitrocellulose membranes with a 0.45 µm pore size (Amersham). Wet blot was performed for 2 h 30 min at 100 V. Membranes were saturated for 1 h with 5% milk (Roth) in TBS with 0.1% Tween-20 (TBST, Sigma-Aldrich) at room temperature.
Primary antibodies Tenascin C (MAB2138, R&D Systems, rat), Collagen-VII alpha 1 (NC2-10, in house, rabbit) and Nidogen-1H-200 (sc-33141, Santa Cruz Biotech, rabbit), were diluted 1:1000 in 5% milk in TBST and used overnight at 4 °C. Membranes were washed three times with TBST, with constant shaking. Secondary antibodies Goat Anti-Rat IgG H + L HRP (31470, Thermo Fisher Scientific) and peroxidase-conjugated Goat Anti-Rabbit (Jackson 111-035-045) were diluted 1:10,000 in 5% milk in TBST and used for 1 h at room temperature. Proteins were revealed by chemiluminescence with SuperSignal West Femto Chemiluminescent (Pierce).

Vu B., Souza G.R, & Dengjel J. (2021). Scaffold-free 3D cell culture of primary skin fibroblasts induces profound changes of the matrisome. Matrix Biology Plus, 11, 100066.

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Immunoblotting with Diverse Antibodies

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The following antibodies were used for immunoblotting: Rat anti-HA (Roche, ROAHAHA), Rabbit anti- FLAG (Proteintech, 20543-1-AP), Mouse anti-GAPDH (Developmental Studies Hybridoma Bank, hGAPDH-2G7), Rabbit anti-USP46 (Proteintech, 13502-1-AP), Mouse anti-Tubulin (Developmental Studies Hybridoma Bank, E7), Mouse anti-β-catenin (BD Transduction Laboratory, 610154), Rabbit anti-Axin1 (Cell Signaling Technology, 2087), Rabbit anti-Insulin Receptor β (Cell Signaling Technology, 3025), Rabbit anti-LRP6 (Cell Signaling Technology, 2560), Rabbit anti-WDR20 (Bethyl Laboratories, A301-657A), Rabbit anti-WDR48 (UAF1) (Proteintech, 16503-1-AP), Goat anti-rat IgG H + L-HRP (Thermo, 31470), Goat anti-mouse IgG H + L-HRP (Promega, W4021), Goat anti-rabbit IgG H + L-HRP (Promega, W4011). All primary antibodies were used at 1:1000 dilution except anti-FLAG (1:2000), anti-GAPDH (1:500), and anti-WDR20 (1:2500). All secondary HRP antibodies were used at 1:5000 dilution.

Ng V.H., Spencer Z., Neitzel L.R., Nayak A., Loberg M.A., Shen C., Kassel S.N., Kroh H.K., An Z., Anthony C.C., Bryant J.M., Lawson A., Goldsmith L., Benchabane H., Hansen A.G., Li J., D’Souza S., Lebensohn A.M., Rohatgi R., Weiss W.A., Weiss V.L., Williams C., Hong C.C., Robbins D.J., Ahmed Y, & Lee E. (2023). The USP46 complex deubiquitylates LRP6 to promote Wnt/β-catenin signaling. Nature Communications, 14, 6173.

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Western Blot Analysis of Apoptosis and Wnt Signaling

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This method was used to semi-quantify the proteins as mentioned below, and cells were harvested and lysed by RIPA Lysis Buffer (Beyotime) to obtain total protein. After that, 40 μg of total protein was measured with BCA protein Assay Kit (Beyotime). Thereafter, proteins were isolated with 8% SDS-PAGE and transferred into Polyvinylidene Fluoride (PVDF) membranes (Invitrogen) later. Next, 8% skimmed milk powder was selected to block membranes for 2 hours and anti-pro-caspase-3 (1:1000; ab183179, Abcam), anti-cleaved-caspase-3 (1:1000; ab49822), anti-p53 (1:1000; ab26), anti-PARP1 (1:1000; ab227224), anti-Wnt3a (1:1000; ab219412), anti-Wnt5a (1:1000; ab227229), anti-β-catenin (1:1000; ab32572) and anti-GAPDH (1:2000; ab181602) were incubated with PVDF membranes at 4°C overnight. Afterwards, membranes were rinsed by Pierce™ Protein-Free T20 (PBS) Blocking Buffer (Thermo Scientific) three times and incubated with secondary antibodies -Goat Anti-Rat IgG H&L (HRP) (ab97057) at room temperature for 2 hours. Finally, BeyoECL Moon (Beyotime) was used for testing protein.

Zhang N, & Wang X. (2020). Circular RNA ITCH mediates H₂O₂-induced myocardial cell apoptosis by targeting miR-17-5p via wnt/β-catenin signalling pathway. International journal of experimental pathology.

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Western Blot Analysis of Protein Extracts

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The cell pellets were lysed in RIPA buffer (150 mM NaCl, 50 mM Tris pH 7.4, 1% Triton X-100, 0.5% NaDoc, 10% glycerol, and 2.5% sodium deoxycholate) with protease and phosphatase inhibitors. Protein concentration was determined using a BCA kit (cat. no. P0010; Beyotime Institute of Biotechnology) and Multiskan™ FC Microplate Reader (Thermo Fisher Scientific Inc.), and analyzed using Skanit software version 3.1 (Thermo Fisher Scientific Inc.). A total of 60 µg of proteins were loaded onto 12% sodium dodecyl sulfide-polyacrylamide gel electrophoresis (SDS-PAGE) for electrophoresis and transferred to polyvinylidene difluoride (PVDF) membranes (Immobilon-P; cat. no. IPVH00010; 0.45 µm; EMD Millipore). Membranes were blocked in 5% milk solution for 2 h at room temperature, and then incubated overnight at 4°C with indicated primary monoclonal antibodies with their respective dilutions (Table SIII). The membranes were then incubated with goat anti-rabbit IgG (H+L)-HRP (1:5,000; cat. no. 31466; Thermo Fisher Scientific Inc.), goat anti-rat IgG (H+L)-HRP (1:5,000; cat. no. 31470; Thermo Fisher Scientific Inc.) and goat anti-mouse IgG (H+L)-HRP (1:5,000; cat. no. 31431; Thermo Fisher Scientific Inc.) for 1 h at room temperature, and blots were visualized with Western Lighting Pro ECL (Fusion FX5; Vilber).

Zhang P., Wang Y., Qin M., Li D., Odhiambo W.O., Yuan M., Lv Z., Liu C., Ma Y., Dong Y, & Ji Y. (2020). Involvement of Blnk and Foxo1 in tumor suppression in BCR-ABL1-transformed pro-B cells. Oncology Reports, 45(2), 693-705.

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