Transblot turbo mini size pvdf membrane

Manufactured by Bio-Rad

Sourced in United States

TransBlot Turbo Mini-size PVDF membranes are a type of laboratory equipment used for protein transfer in western blotting applications. They are made of polyvinylidene fluoride (PVDF) material and are designed for use with the TransBlot Turbo transfer system.

Automatically generated - may contain errors

Lab products found in correlation

Mini protean tgx stain free gels 4 20, by Bio-Rad (2 mentions) Odyssey fc system, by LI COR (1 mentions) Ab214428, by Abcam (1 mentions) Supersignal west pico plus chemiluminescent substrate, by Thermo Fisher Scientific (1 mentions) Donkey antisheep hrp, by Thermo Fisher Scientific (1 mentions) Af2557, by R&D Systems (1 mentions) Supersignal west femto maximum sensitivity substrate, by Thermo Fisher Scientific (1 mentions) Clarity western ecl blotting substrate, by Bio-Rad (1 mentions) Gradient polyacrylamide gel, by Bio-Rad (1 mentions) Blotting grade blocker, by Bio-Rad (1 mentions) Psd 95, by Thermo Fisher Scientific (1 mentions) Trans blot turbo system, by Bio-Rad (1 mentions) Anti collagen 1 and collagen 3, by Abcam (1 mentions) Anti mmp 12, by Santa Cruz Biotechnology (1 mentions) Anti elastin, by Santa Cruz Biotechnology (1 mentions) Bullet blocking one, by Nacalai Tesque (1 mentions) β actin monoclonal antibody, by Cell Signaling Technology (1 mentions) Chemidoc mp image lab, by Bio-Rad (1 mentions) Horseradish peroxidase hrp conjugated anti rabbit antibody, by GE Healthcare (1 mentions) Clarity western ecl substrate, by Bio-Rad (1 mentions)

5 protocols using transblot turbo mini size pvdf membrane

Autophagy Inhibition Impacts Protein Labeling

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

MEFs were plated in a 12-well plate at a density of 150,000 cells per well and incubated overnight at 37° C. The next day, two hours prior to adding the BMV109 (Vergent Bioscience 40200–200) probe, selected wells were treated with a final concentration of 40 nM Bafilomycin A1(Fermentek 88899-55-2) in complete media to inhibit autophagosome-lysosome fusion in control conditions. The probe was administered at a final concentration of 1 μM in media for experimental conditions and a final concentration of 1 μM BMV109 and 40 nM Bafilomycin A1 in control conditions and cells were incubated for one hour. Proteins were resolved by Mini-Protean TGX stain-free gels 4–20% (BioRad 4568093) and transferred to TransBlot Turbo mini-size PVDF membrane (BioRad). Membranes were fixed with 0.4% PFA then rinsed three times with water. BMV109 signal was detected using the Li-Cor Odyssey Fc system.

Anderson A, & Tansey M.G. (2023). Loss of Progranulin Results in Increased Pan-Cathepsin Activity and Reduced LAMP1 Lysosomal Protein. bioRxiv.

+ Open protocol

+ Expand

Western Blot Analysis of Lysosomal Proteins

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

Proteins were resolved by Mini-Protean TGX stain-free gels 4–20% (BioRad 4568093) and transferred to TransBlot Turbo mini-size PVDF membrane (BioRad). Membranes were fixed with 0.4% paraformaldehyde (PFA) then rinsed three times with ultrapure water using the Milli-Q EQ 7000 Ultrapure Water Purification System. The membrane was blocked in 5% powdered milk in 1X tris-buffered saline with Tween-20 (TBS-T), and primary antibodies and secondary antibodies were dissolved in this same buffer. Information on the antibodies used can be found in Table 1. Primary antibodies used were LAMP1 (rabbit, Abcam ab24170) diluted 1:1000, PGRN (sheep, R&D AF2557) diluted 1:400 and Cathepsin B (CTS B)(rabbit, Abcam ab214428) diluted 1:1000. Secondary antibodies Goat anti Rabbit HRP (Jackson Immunoresearch 111-035-144) and Donkey anti-Sheep HRP (Invitrogen A16041) were diluted 1:2000. The membrane was developed using SuperSignal West Pico PLUS Chemiluminescent Substrate (Thermo scientific 34577) and SuperSignal West Femto Maximum Sensitivity Substrate (Thermo scientific 34096) and visualized on the Li-cor Odyssey FX system.

Anderson A, & Tansey M.G. (2023). Loss of Progranulin Results in Increased Pan-Cathepsin Activity and Reduced LAMP1 Lysosomal Protein. bioRxiv.

+ Open protocol

+ Expand

Protein Quantification and Western Blot Analysis

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

Protein concentrations were measured with the BCA assay (Pierce, Thermo Scientific, 23225) before SDS-PAGE and Western blot analysis. All protein samples were subjected to SDS-PAGE using 4–15% polyacrylamide gradient gels (Bio-Rad, 4561086). For cell lysates, 20–30 μg was loaded, while for synaptosomal fraction samples 40 μg of protein for each sample was loaded on the gels. Separated proteins were then electroblotted using a TransBlot Turbo system to TransBlot Turbo Mini-size PVDF membranes (Bio-Rad, 1704272). Membranes were subsequently blocked with 1× TBS containing 0.1% Tween 20 (1× TBST) with 5% (w/v) blotting-grade blocker (Bio-Rad, 1706404) and probed with the primary antibody of interest diluted in 1× TBS containing 0.1% Tween 20 and 0.5% blocker at room temperature overnight. Primary antibodies for mouse Csmd2 were used at a dilution of 1:500, PSD-95 (ThermoFisher, RRID: AB_325399) at 1:1000, and DYKDDDK (FLAG; ThermoFisher, RRID: AB_2536846) at 1:500. Membranes were washed three times in 1× TBST for 10 min each before 1 h of incubation at room temperature with horseradish peroxidase (HRP)-conjugated secondary antibodies used at 1:10 000. Membranes were visualized using the Clarity Western ECL Blotting Substrates (Bio-Rad, 1705060) according to the manufacturer’s recommended instructions in a Bio-Rad Chemidoc Universal Hood III imaging system.

Gutierrez M.A., Dwyer B.E, & Franco S.J. (2019). Csmd2 Is a Synaptic Transmembrane Protein that Interacts with PSD-95 and Is Required for Neuronal Maturation. eNeuro, 6(2), ENEURO.0434-18.2019.

+ Open protocol

+ Expand

Quantifying Arterial Extracellular Matrix Proteins

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

Arterial tissue (carotid arteries and thoracic aorta) was collected and snap-frozen in liquid nitrogen before homogenization in modified RIPA lysis buffer containing 1% SDS and 1% Triton. For collagen extraction, tissue was first subjected to acid hydrolysis with 6N HCl at 100°C for 72 hours, then homogenized in RIPA lysis buffer containing 1% SDS and 1% Triton. Equal amounts of protein were separated by SDS-PAGE and transferred to Trans-Blot® Turbo^™ Mini-size PVDF membranes (Biorad, Hercules, CA). Primary anti-CYP4A1 (1:200, Santa Cruz, Dallas, TX), anti-MMP12 (1:250, Santa Cruz, Dallas, TX), anti-MMP2 and -MMP9 (1:1000, Millipore, Temecula, CA), anti-MMP3, -MMP7 and – MT1-MMP (1:1000, Abcam, Cambridge, MA), anti-TIMP1, -TIMP2, -TIMP3 and – TIMP4 (1:1000, Abcam), anti-collagen I and – collagen III (1:1,000, Abcam), and anti-elastin (1:200, Santa Cruz, sc-58756, recognizes intact elastin and degradation products) and secondary anti-rabbit or anti-mouse antibodies (BioRad), as appropriate, were used for Western blotting. Bands were visualized by the Odyssey Western blot detection system (Azure Biosystems, Dublin, CA). Bands were quantified using Un-Scan-It Image software (Silk Scientific Corporation, Orem, UT). Data are normalized to β-tubulin (loading control).

Soler A., Hunter I., Joseph G., Hutcheson R., Hutcheson B., Yang J., Zhang F.F., Joshi S.R., Bradford C., Gotlinger K.H., Maniyar R., Falck J.R., Proctor S., Schwartzman M.L., Gupte S.A, & Rocic P. (2018). Elevated 20-HETE in Metabolic Syndrome Regulates Arterial Stiffness and Systolic Hypertension via MMP12 Activation. Journal of molecular and cellular cardiology, 117, 88-99.

+ 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

We used an SDS buffer (25% of 0.125M Tris-HCl (pH 6.8), 20% of glycerol, 4% of SDS, and 10% of 2-mercaptoethanol, with bromophenol blue included) to pellet cell pellets, extract proteins, and degrade DNA by sonication in ice-cold conditions. The protein extracts were separated in Mini-PROTEAN TGX Stain-Free gels (BIO RAD, CA, USA) by electrophoresis (60 mA, 40 min) and transferred to TransBlot Turbo Mini-size PVDF membranes (BIO RAD). The membranes were blocked with Bullet Blocking One (Nacalai Tesque, Kyoto, Japan). The antibodies we used were as follows: primary antibodies—SHC4 polyclonal antibody (rabbit, 75 kDa) (Sigma-Aldrich, Mizulli, USA) and β-actin monoclonal antibody (rabbit; 45 kDa) (Cell signaling technology, #4790, MA, USA) diluted 1000×, reacted at 4 °C in overnight. The membranes were washed with TBS-T, and the horseradish peroxidase (HRP)-conjugated anti-rabbit antibody (GE Healthcare, Chicago, IL, USA) diluted 5000× was reacted with membrane for 1 h. The membranes were washed with TBS-T three times before being exposed to the ClarityTM Western ECL Substrate (Bio Rad Laboratories, Hercules, CA, USA) as a chromogenic substrate for HRP. The protein bands were detected using a ChemiDoc MP ImageLab (Bio Rad Laboratories, Hercules, CA, USA).

Koyanagi A., Onishi I., Muraoka K., Sato I., Sato S., Kimura T., Kishida A., Yamamoto K., Kitagawa M, & Kurata M. (2022). Identification of the Factor That Leads Human Mesenchymal Stem Cell Lines into Decellularized Bone. Bioengineering, 9(10), 490.

+ 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!