Transfer blot turbo transfer system

Manufactured by Bio-Rad

Sourced in United States

The Transfer-Blot Turbo Transfer System is a laboratory equipment designed for rapid and efficient protein transfer from polyacrylamide gels to membranes. It utilizes a specialized transfer buffer and high-powered electrical current to facilitate the transfer process, enabling researchers to perform western blotting analyses in a timely manner.

Automatically generated - may contain errors

Lab products found in correlation

9 protocols using transfer blot turbo transfer system

Western Blot Analysis of Proteins

Check if the same lab product or an alternative is used in the 5 most similar protocols

The protein samples were separated on 7.5% tris-glycine polyacrylamide gels (Bio-Rad) (SPT16, XRN2, SETDB1, ATRX, and DNMT3B) or 12% polyacrylamide gels [telomere repeat binding factor 1 (TRF1), TRF2, histones, and their modifications]. Proteins were transferred to nitrocellulose membranes using the transfer-blot Turbo Transfer system (Bio-Rad). Primary and secondary horseradish peroxidase–conjugated antibodies were incubated by standard Western blotting procedures and detected by adding ECL chemiluminescent solution (PerkinElmer). Signals were captured on autoradiography films (Amersham).

Gauchier M., Kan S., Barral A., Sauzet S., Agirre E., Bonnell E., Saksouk N., Barth T.K., Ide S., Urbach S., Wellinger R.J., Luco R.F., Imhof A, & Déjardin J. (2019). SETDB1-dependent heterochromatin stimulates alternative lengthening of telomeres. Science Advances, 5(5), eaav3673.

+ Open protocol

+ Expand

Hippocampal Protein Expression Analysis

Check if the same lab product or an alternative is used in the 5 most similar protocols

Hippocampus samples (30 μg protein/well) and a marker protein (Bio-Rad, São Paulo, Brazil) were separated on an SDS–polyacrylamide gel by electrophoresis. The proteins were transferred to a nitrocellulose membrane (0.45 μm, Bio-rad) using the Transfer-Blot® Turbo ™ transfer system (1.0 mA, 15 to 40 min, Bio-Rad). After blocking with a 3% bovine serum albumin (BSA) solution for 1 h, the blots were incubated overnight at 4 °C with primary antibodies (Table S1). The method was performed as previously described by Müller et al. [28 ].

Jung J.T., Marques L.S., Zborowski V.A., Silva G.L., Nogueira C.W, & Zeni G. (2022). Resistance Training Modulates Hippocampal Neuroinflammation and Protects Anxiety-Depression-like Dyad Induced by an Emotional Single Prolonged Stress Model. Molecular Neurobiology, 60(1), 264-276.

+ Open protocol

+ Expand

GFP-Trap Based Protein Purification

Check if the same lab product or an alternative is used in the 5 most similar protocols

150 mm dishes of near-confluent cells were treated as indicated, washed with cold Hanks Basic Salt Solution, and scrape-harvested at 4°C into lysis buffer (20 mM Tris-HCl pH 7.4, 150 mM NaCl, 5 mM MgCl2, 1 mM DTT 0.5% NP-40, 10% glycerol) containing 1 mM DTT, protease inhibitors (Roche, EDTA free), HALT phosphatase inhibitors (Pierce), and 20 μg/nL RNase A. Cells were rotated for 30-minutes at 4°C, cleared by centrifugation (5000 rpm for 5-minutes), and supernatants removed then incubated with Chromotek-GFP-Trap® Beads (Allele Biotech) for 2-hours with continuous rotation at 4°C. Beads were washed 5-times, and either eluted directly into SDS-lysis buffer with RNase treatment, or extracted in RIPA buffer (50 mM TRIS, 150 mM NaCl, 1.0% NP40, 0.5% DOC, 0.05% SDS) for 1-hour at 4°C with rotation. Material released by RIPA buffer was recovered and precipitated with 60% acetone. Beads post-RIPA extraction contained bound material denoted “high-affinity”, which was released by heating in reducing SDS-PAGE lysis buffer. Proteins were resolved on 4–20% Mini-PROTEAN TGX Precast Gel (Bio-Rad), transferred to nitrocellulose membranes using the Transfer-Blot Turbo transfer system (Bio-Rad), and blotted using standard procedures as above. Chemiluminescence was detected using SuperSignal West Pico substrate (Thermo Scientific).

Sanders D.W., Kedersha N., Lee D.S., Strom A.R., Drake V., Riback J.A., Bracha D., Eeftens J.M., Iwanicki A., Wang A., Wei M.T., Whitney G., Lyons S.M., Anderson P., Jacobs W.M., Ivanov P, & Brangwynne C.P. (2020). Competing protein-RNA interaction networks control multiphase intracellular organization. Cell, 181(2), 306-324.e28.

+ Open protocol

+ Expand

Western Blot Analysis of HLTF and Heparanase

Check if the same lab product or an alternative is used in the 5 most similar protocols

Pulled down proteins were separated on Mini-PROTEAN^® TGX^TM 4–20% precast gels (BIO-RAD Laboratories, Inc. USA) and then transferred onto a 0.2 µm nitrocellulose membrane by Trans-Blot^® Turbo^TM Transfer Pack (BIO-RAD Laboratories, Inc. USA) and Transfer Blot Turbo Transfer System (BIO-RAD). Each membrane was blocked with 5% BSA in PBS-T. A rabbit-anti-HLTF antibody (Abcam, ab17984) and rabbit-anti-heparanase antibody (InSight Pharmaceuticals, Rehovot, Israel) were applied for the assay. After washing, membrane was incubated with HRP-conjugated anti-rabbit secondary antibody. Detection was performed by applying enhanced chemiluminescence (EZ-ECL, Biological Industries, Israel).

Ostrovsky O., Grushchenko-Polaq A.H., Beider K., Mayorov M., Canaani J., Shimoni A., Vlodavsky I, & Nagler A. (2018). Identification of strong intron enhancer in the heparanase gene: effect of functional rs4693608 variant on HPSE enhancer activity in hematological and solid malignancies. Oncogenesis, 7(6), 51.

+ Open protocol

+ Expand

Western Blot Analysis of Protein Lysates

Check if the same lab product or an alternative is used in the 5 most similar protocols

Whole-cell lysates from HeLa and SH-SY5Y were prepared using ice-cold radioimmunoprecipitation assay (RIPA) buffer (Boston BioProducts; See Supplementary Experimental Procedures). Protein samples were resolved on 11 or 12% SDS-polyacrylamide gels, transferred to membranes (Immun-Blot PVDF Membrane for Protein Blotting, Bio-Rad) using Transfer-Blot Turbo Transfer System (Bio-Rad), and blocked with 5% nonfat dried milk in Tris-buffered saline containing 0.05% Tween-20 (TBST). Antibody dilutions and incubation conditions are given in Supplementary Methods. Proteins were visualized using Clarity Western ECL Blotting substrate (Bio-Rad), SuperSignal West Dura Extended Duration Substrate or SuperSignal West Femto Maximum Sensitivity Substrate (Thermo Scientific). The membranes were scanned using a UVP BioSpectrum AC Imaging System (UVP). Densitometry measurements were done using ImageJ software. Results were confirmed by at least two independent experiments.

Seo J., Singh N.N., Ottesen E.W., Lee B.M, & Singh R.N. (2016). A novel human-specific splice isoform alters the critical C-terminus of Survival Motor Neuron protein. Scientific Reports, 6, 30778.

+ Open protocol

+ Expand

P-glycoprotein Expression Analysis by Western Blot

Check if the same lab product or an alternative is used in the 5 most similar protocols

Western Blot analysis was used to assess the protein expression levels and was performed as previously described (Tresch et al., 2021 ). In brief, cell lysates were generated using RIPA lysis buffer and protein concentration was measured with the Pierce™ Protein Assay Kit (Thermo Fischer, Waltham, MA, USA). Total protein of 50 μg was separated by SDS‐Page on a 4–15% gradient gel (Bio‐Rad) and transferred on a PVDF membrane using a transfer apparatus (Transfer‐Blot Turbo Transfer System, Bio‐Rad, Hercules, CA, USA). Blotting membranes were blocked with 5% nonfat milk in TBST (Bio‐Rad, Hercules, CA, USA), incubated overnight with an antibody against P‐gp (C219, #MA1‐26528, 1:1000 Invitrogen, Waltham, MA, USA) and detected following incubation with an anti‐mouse IgG, HRP‐linked secondary antibody (#7076S, 1:1000, Cell Signaling). For loading control, a β‐actin antibody (8226, 1:1000, Abcam) was used. Pierce™ ECL Western Blotting Substrate (Thermo Fischer Scientific, Waltham, MA, USA) was used for chemiluminescent detection with the Fusion Solo S Edge – Chemiluminescence Imaging System (Witec AG, Sursee, Switzerland).

Fuchs D., Rohrer Bley C., Morandi L., Tonon C., Weyland M.S, & Nytko K.J. (2023). Triple combination of lomustine, temozolomide and irradiation reduces canine glioma cell survival in vitro. Veterinary Medicine and Science, 9(4), 1573-1583.

+ Open protocol

+ Expand

BV2 Cell Lysis and Western Blot Analysis

Check if the same lab product or an alternative is used in the 5 most similar protocols

BV2 cells were lysed in RIPA buffer and determined with Bradford reagent (Beyotime, cat. no. P0006C). The protein samples were separated on 10% tris-glycine polyacrylamide gels (Sangon Biotech, China, cat. no. C681102). Transfer proteins to nitrocellulose membranes with the Transfer-Blot Turbo Transfer System (Bio-Rad, USA). Membranes were blocked with Protein-Free Rapid Blocking Buffer (Epizyme Biotech, China, cat. no. PS108P) for 15 min and incubated overnight with primary antibodies. The following primary antibodies were used: anti-tubulin antibody (1:1,000; ABclonal, China, cat. no. AC026) and anti-IL-1β (1:1,000; RD, USA, cat. no. AF-401). Tubulin was detected using goat anti-rabbit secondary antibody (1:5,000; Jackson Immuno Research Labs, USA, cat. no. 111-035-003), and IL-1β was detected using donkey anti-goat secondary antibody (1:2,000; Beyotime, China, cat. no. A0181). Signals were visualized with the ChemiDoc Imaging System (Bio-rad).

Lv K., Ying H., Hu G., Hu J., Jian Q, & Zhang F. (2022). Integrated multi-omics reveals the activated retinal microglia with intracellular metabolic reprogramming contributes to inflammation in STZ-induced early diabetic retinopathy. Frontiers in Immunology, 13, 942768.

+ Open protocol

+ Expand

Protein Extraction and Western Blot Analysis

Check if the same lab product or an alternative is used in the 5 most similar protocols

Cells were lysed in RIPA buffer containing protease and phosphatase inhibitor cocktails, along with 1 mM phenylmethylsulfonyl fluoride. After addition of lysis buffer, cells were placed on ice for 20 min and then sonicate for 5 min. Cell lysates were centrifuged at 13,000× g for 10 min at 4 °C. Protein was quantified using BCA kit (cat. no. 23225, ThermoFisher Scientific). Next, 10 µg protein samples were separated on 10% SDS PAGE gels. Proteins were transferred onto nitrocellulose membrane (cat. no. NBA 083C001EA, Protran) using a wet transfer system or semidry Transferblot Turbo Transfer System (Bio-Rad, Hercules, CA, USA). The transferred proteins were membrane blocked in 5% skimmed milk for 30 min. The membrane was incubated with primary antibody at a 1:2000 dilution in 5% BSA overnight at 4 °C. The next day, membranes were washed for 3 × 10 min and then incubated with HRP-conjugated secondary antibody for 1 h at room temperature. After the ECL-reaction (cat. no. 170-5061, BioRad), protein bands were quantified using a ChemiDoc MP instrument (Bio-Rad). All of the antigens and molecular weights in kDa are labelled next to the blots on the figures.

Siddiqui F.A., Vukic V., Salminen T.A, & Abankwa D. (2021). Elaiophylin Is a Potent Hsp90/ Cdc37 Protein Interface Inhibitor with K-Ras Nanocluster Selectivity. Biomolecules, 11(6), 836.

+ Open protocol

+ Expand

Western Blot Protein Detection Protocol

Check if the same lab product or an alternative is used in the 5 most similar protocols

The proteins were transferred onto a 0.2 μm PVDF membrane for 10 min at 25 V and 1.3 A using the Transfer-Blot Turbo Transfer System (BioRad). For Western blot analysis the iBind Western System (Novex) was used. The membrane was incubated with the monoclonal mouse anti-His-Tag antibody (Novagen) and WesternSure HRP-conjugated goat anti-mouse IgG (Li-COR) for 2.5 h at room temperature. The detection was performed by incubating the membrane for 5 min in the WesternSure Premium Chemiluminescent Substrate (Li-COR) and scanning the blot using C-DiGit Blot Scanner (Li-COR).

Kutta R.J., Archipowa N., Johannissen L.O., Jones A.R, & Scrutton N.S. (2017). Vertebrate Cryptochromes are Vestigial Flavoproteins. Scientific Reports, 7, 44906.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!