Vitrobot filter paper

Manufactured by Ted Pella

Sourced in United States

The Vitrobot filter paper is a specialized laboratory equipment used for sample preparation in cryo-electron microscopy. It is designed to absorb excess liquid from samples, facilitating the formation of thin, uniform samples suitable for vitrification and imaging. The filter paper functions by wicking away excess liquid from the sample, helping to control the thickness of the sample and improve the quality of the resulting cryo-EM images.

Automatically generated - may contain errors

Lab products found in correlation

Vitrobot mark 4, by Thermo Fisher Scientific (5 mentions) Lacey carbon grid, by Ted Pella (2 mentions) Vitrobot plunge freezer, by Thermo Fisher Scientific (2 mentions) Talos arctica 200 kv transmission electron microscope, by Thermo Fisher Scientific (2 mentions) Gatan k3 camera, by Ametek (2 mentions) Superdex 200 increase 10 300 column, by GE Healthcare (1 mentions) Ultrascan 1000xp ccd camera, by Ametek (1 mentions) Titan krios g3 electron microscope, by Thermo Fisher Scientific (1 mentions) Ultraufoil, by Quantifoil (1 mentions) Igepal ca 630, by Merck Group (1 mentions) Atpγs, by Merck Group (1 mentions)

Spelling variants of this product

Filter paper (18 citations) Vitrobot filter paper (grade 595, (4 citations) Standard Vitrobot Filter Paper (3 citations)

8 protocols using vitrobot filter paper

Cryo-EM Imaging of Extracellular Vesicles

Cited 1 time

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

Three to four microliters of the EV solution (1:20 dilution) was added to Lacey carbon grids (300-mesh; Ted Pella, Inc., Redding, CA, USA) that were negatively glow-discharged for 80 s at 30 mA. Excess sample was removed by blotting once for 3 s with Vitrobot filter paper (Ted Pella, Inc, Redding, CA, USA) and then the grid was plunge-frozen in liquid ethane cooled by liquid nitrogen using a Vitrobot plunge-freezer (ThermoFisher Scientific, Hillsboro, OR, USA).
The vitrified vesicle samples were imaged using a Talos Arctica 200 kV transmission electron microscope (ThermoFisher Scientific, Hillsboro, OR, USA) equipped with a Gatan K3 camera (Gatan, Inc., Pleasanton, CA, USA) The SerialEM software was used to collect images under low-dose conditions at 36,000 × magnification corresponding to a pixel size of 1.14 Å/pixel. For each image, 50 frames were recorded over 2.5 s exposure time at a dose rate of 35 electrons/pixel/s. The movie frames were aligned using MotionCorr2 (2) under Relion (Zivanov et al. 2018 (link)).

Adduri R.S., Vasireddy R., Mroz M.M., Bhakta A., Li Y., Chen Z., Miller J.W., Velasco-Alzate K.Y., Gopalakrishnan V., Maier L.A., Li L, & Konduru N.V. (2022). Realistic biomarkers from plasma extracellular vesicles for detection of beryllium exposure. International Archives of Occupational and Environmental Health, 95(8), 1785-1796.

+ Open protocol

+ Expand

Cryo-EM Imaging of Extracellular Vesicles

Cited 1 time

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

+ Open protocol

+ Expand

AsCas12f Protein-RNA-DNA Complex Preparation

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

The AsCas12f-sgRNA-DNA complex was assembled by mixing purified AsCas12f (D225A), the 194 nt sgRNA, the 42 nt target DNA, and the 42 nt non-target DNA, at a molar ratio of 1:0.5:1.2:1.2. Sequences of the sgRNA and the target DNA are provided in Supplementary Table 1. The mixture was incubated on ice for 30 min before being loaded onto a Superdex 200 Increase 10/300 column (GE Healthcare) equilibrated with buffer D (50 mM Tris-HCl, pH 8.0, 50 mM NaCl, 5 mM MgCl₂, and 0.5 mM TCEP). Fractions that contain the pure AsCas12f-sgRNA-DNA complex were pooled and concentrated to roughly 2.5 mg/mL.
Sample vitrification was performed using a Vitrobot Mark IV (Thermo Fisher) operating at 8 °C and 100% humidity. 3.5 μL sample was applied to holey carbon grids (Quantifoil 200 mesh Cu 1.2/1.3) that had been glow-discharged for 30 seconds. The grids were blotted for 4 seconds at a “blotting force” 0 by standard Vitrobot filter paper (Ted Pella, 47000–100) and were then plunge-frozen in liquid ethane.

Delphin C., Guan T., Melchior F, & Gerace L. (1997). RanGTP Targets p97 to RanBP2, a Filamentous Protein Localized at the Cytoplasmic Periphery of the Nuclear Pore Complex. Molecular Biology of the Cell, 8(12), 2379-2390.

+ Open protocol

+ Expand

Cryo-EM Grid Preparation for CA

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

Cryo-EM grids of CA were prepared using Vitrobot Mark IV (Thermo Fisher Scientific). Three microliters of the purified CA sample was applied to each Quantifoil holy carbon grid (R2/1, 300 mesh gold). All grids were incubated for 5 seconds in the Vitrobot chamber at 8 °C and 100% humidity, blotted with standard Vitrobot filter paper (Ted Pella), and then plunged into liquid ethane at approximately −170 °C.

Han L., Rao Q., Yang R., Wang Y., Chai P., Xiong Y, & Zhang K. (2022). Cryo-EM structure of an active central apparatus. Nature Structural & Molecular Biology, 29(5), 472-482.

+ Open protocol

+ Expand

Cryo-EM Preparation of LDL Nanoparticles

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

LDL nanoparticle samples were prepared for cryo-electron microscopy (cryo-EM) by loading 2.5 μL of sample onto a carbon coated glow-discharged copper grid (Quantifoil grids R2/2, Electron Microscopy Science, Hatfield, PA). After a 60 second incubation the gird was mounted inside the Mark III Vitrobot chamber (FEI USA, Hillsboro, Oregon, USA), which was maintained at 4°C with greater than 90% humidity. The grid was blotted for 7 seconds using a standard Vitrobot Filter paper (Ø55/20 mm, Grade 595, Ted Pella, Inc, Redding, CA), immediately plunged into liquid ethane and stored in liquid nitrogen until EM examination. For cryoEM imaging, the grids were transferred into a JEOL JEM 2200FS Transmission Electron Microscope (TEM; JEOL USA, Inc., Peabody, MA) operating at 200 kV. A 30 eV energy filter was used for zero-loss imaging. Images were exposed with an electron dose of ~20 electrons per Å² and at defocus levels varying from −1.5 to −2.5 μm. Lastly, images were digitized directly with a Gatan UltraScan 1000XP CCD Camera at 25K magnification (Gatan, Inc., Pleasanton, CA).

Mulik R.S., Zheng H., Pichumani K., Ratnakar J., Jiang Q.X, & Corbin I.R. (2017). Elucidating the Structural Organization of a Novel Low-Density Lipoprotein Nanoparticle Reconstituted with Docosahexaenoic Acid. Chemistry and physics of lipids, 204, 65-75.

+ Open protocol

+ Expand

Cryo-EM Structure of PhuR Complex

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

APol-reconstituted native PhuR complexes with sAB11 and the nanobody were concentrated to 2–5 mg/mL using a 100-kDa molecular weight cutoff concentration (Amicon). Prior to vitrification, 0.25% CHAPSO was added to improve the quality of vitreous ice during freezing and reduce denaturation and aggregation at the air-water interface. 3.5 µL of sample was applied to plasma-cleaned (Gatan Solarus) 1.2/1.3-µm UltrAuFoil grids (Quantifoil) and blotted using standard Vitrobot filter paper (Ted Pella, 47000–100) on both sides for 4 seconds using a Vitrobot Mark IV (Thermo Fisher Scientific) operating at 8°C and 100% humidity. The grids were immediately plunged after blotting into liquid ethane for vitrification. Cryo-EM images were collected at the University of Chicago Advanced Electron Microscopy Facility using a Titan Krios G3 electron microscope (Thermo Fisher Scientific) operating at 300 kV and equipped with a Gatan K3 direct detection camera and Energy Filter (BioQuantum). Automated data collection was done in CDS mode using the EPU software package. A total of 5,031 movies consisting of 50 fractions with a 60 e^-/Å total exposure were collected with a super-resolution pixel size of 0.534 Å, a calibrated pixel size of 1.068 Å, and a defocus range of −1.0 to −2.5 µm.

Knejski P.P., Erramilli S.K, & Kossiakoff A.A. (2023). Chaperone-assisted cryo-EM structure of P. aeruginosa PhuR reveals molecular basis for heme uptake. bioRxiv.

+ Open protocol

+ Expand

Cryo-EM Sample Preparation for p97-Npl4/Ufd1 Complex

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

For p97-Npl4/Ufd1 complex, before preparing grids for cryo-EM, the complex was concentrated to ~20 mg/mL, and incubated with 5 mM ATPγS (90% pure, Sigma) for 30 min at room temperature. For p97 (A232E/E578Q)-Npl4/Ufd1-Ub-Eos complex, the sample was concentrated to 20 mg/mL without adding additional nucleotide. For p97-Npl4/Ufd1 complex in the presence of cupric ion, the complex was concentrated to ~20 mg/mL, and incubated with 5 mM ATPγS (90% pure, Sigma) and 100 μM cupric ion for 30 min at room temperature. To relief the preferred orientations, IGEPAL CA-630 (Sigma) was added to the samples to a final concentration of 0.05% immediately before grid freezing. The freezing was performed using a Vitrobot mark IV (Thermo Fisher) operating at 8 °C and 100% humidity. Samples (3.5 μL) were applied to a non-glow-discharged Quantifoil Au 1.2/1.3 grid. The grid was blotted for 1 s using standard Vitrobot filter paper (Ted Pella, 47000-100) and then plunge frozen in liquid ethane.

Pan M., Zheng Q., Yu Y., Ai H., Xie Y., Zeng X., Wang C., Liu L, & Zhao M. (2021). Seesaw conformations of Npl4 in the human p97 complex and the inhibitory mechanism of a disulfiram derivative. Nature Communications, 12, 121.

+ Open protocol

+ Expand

Cryo-EM Grid Preparation for CA

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

Cryo-EM grids of CA were prepared using Vitrobot Mark IV (Thermo Fisher Scientific). Three microliters of the purified CA sample were applied to each Quantifoil holy carbon grid (R2/1, 300 mesh gold). All grids were incubated for 5 s in the Vitrobot chamber at 8 °C and 100% humidity, blotted with standard Vitrobot filter paper (Ted Pella, Inc.), and then plunged into liquid ethane at approximately -170 °C.

Han L., Rao Q., Yang R., Wang Y., Chai P., Xiong Y., & Zhang K. (2022). Cryo-EM structure of an active central apparatus.

+ 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!