Log In Sign up
The largest database of trusted experimental protocols

Pvdf fl membrane

Manufactured by Merck Group
Visit Supplier
Sourced in United States

PVDF-FL membranes are a type of laboratory equipment used for various filtration and separation applications. They are made of polyvinylidene fluoride (PVDF), a durable and chemically resistant material. PVDF-FL membranes are designed to provide efficient and reliable performance in a range of laboratory settings.

Automatically generated - may contain errors

Lab products found in correlation

Odyssey infrared imaging system, by LI COR (9 mentions) Odyssey blocking buffer, by LI COR (5 mentions) Protease inhibitor cocktail, by Merck Group (4 mentions) Odyssey imaging system, by LI COR (3 mentions) Irdye800 conjugated goat anti mouse, by LI COR (2 mentions) Odyssey imager, by LI COR (2 mentions) Novex nupage lds sample buffer, by Thermo Fisher Scientific (2 mentions) Nupage bis tris gel, by Thermo Fisher Scientific (2 mentions) Ab22553, by Abcam (2 mentions) Af 1655, by R&D Systems (2 mentions) Ab204448, by BD (2 mentions) A21057, by Thermo Fisher Scientific (2 mentions) Odyssey image studio software, by LI COR (2 mentions) Odyssey instrument, by LI COR (2 mentions) α actin, by Cell Signaling Technology (2 mentions) α rabbit hrp, by Cell Signaling Technology (2 mentions) αrabbit irdye 800cw, by LI COR (2 mentions) αmouse irdye 680lt, by LI COR (2 mentions) α mouse hrp, by Cell Signaling Technology (2 mentions) Irdye 800cw goat anti rabbit, by LI COR (2 mentions)

Close spelling variants of this product

Immobilon-FL transfer PVDF membranes (4 citations) Immobilon-FL 0.45 µm PVDF membranes (2 citations) Immobilon-FL polyvinylidene fluoride (PVDF) membranes (6 citations) Mobilon-FL PVDF membranes (5 citations) Immobilin-FL PVDF membranes (2 citations) Immobilion-FL PVDF membranes (5 citations) Immunobilon-FL PVDF membrane (2 citations) IMMOBILON-FL 26 PVDF membrane (2 citations) Immobilon-FL PVDF transfer membranes (9 citations) Immobilon‐FL polyvinylidene difluoride (PVDF) membrane (14 citations)

46 protocols using pvdf fl membrane

1

Western Blot Analysis of DPD Protein

Check if the same lab product or an alternative is used in the 5 most similar protocols
Protein lysates were separated by SDS-PAGE and transferred to polyvinylidene difluoride (PVDF)-FL membrane (EMD Millipore, Saint Charles, MO). Membranes were blocked using Odyssey blocking buffer (LI-COR Biosciences, Lincoln, NE). Blots were probed with primary antibodies against DPD and α-tubulin (ab6556 and ab186407, respectively; Abcam, Cambridge, MA). IRDye800-conjugated goat anti-mouse and IRDye680-conjugated goat anti-rabbit secondary antibodies (LI-COR) were used for detection. Blots were scanned on a LI-COR Odyssey imager.
Nie Q., Shrestha S., Tapper E.E., Trogstad-Isaacson C.S., Bouchonville K.J., Lee A.M., Wu R., Jerde C.R., Wang Z., Kubica P.A., Offer S.M, & Diasio R.B. (2017). Quantitative contribution of rs75017182 to dihydropyrimidine dehydrogenase mRNA splicing and enzyme activity. Clinical pharmacology and therapeutics, 102(4), 662-670.
+ Open protocol
+ Expand
2

Mitochondrial Protein Complex Analysis

Cited 1 time
Check if the same lab product or an alternative is used in the 5 most similar protocols
The mitochondrial fraction was solubilized with 5% (w/v) digitonin (Invitrogen)31 (link),32 (link) for 15 min on ice. After centrifugation at 20,000 × g for 30 min at 4 °C, the clear supernatant was loaded onto 3–12% BN gels (Invitrogen). For the immunoblot analysis, proteins in the gels were transferred onto a PVDF-FL membrane (Merck). The in-gel CcO activity of CIV was visualized by the precipitation of 3, 3′-diaminobenzidine oxides and indamine polymers in test buffer containing 50 mM sodium phosphate, 0.5 mg/ml 3, 3′-diaminobenzidine-tetrahydrochloride (D5905; Sigma), 0.25 mg/l cyt c (C2037; Sigma), and 75 mg/ml sucrose, pH 7.432 (link).
Takahashi K., Miura Y., Ohsawa I., Shirasawa T, & Takahashi M. (2018). In vitro rejuvenation of brain mitochondria by the inhibition of actin polymerization. Scientific Reports, 8, 15585.
+ Open protocol
+ Expand
3

Western Blot Protein Detection

Check if the same lab product or an alternative is used in the 5 most similar protocols
Protein lysates were separated on a 10% SDS-PAGE gel and transferred to PVDF-FL membrane (EMD Millipore). Membranes were blocked using Odyssey blocking buffer (LI-COR, Lincoln, NE). Blots were probed with primary antibodies against DPD and alpha-tubulin (both AbCam, Cambridge, MA) and subsequent secondary IRDye800 conjugated goat anti-mouse and IRDye 680 conjugated goat anti-rabbit (both LI-COR). Blots were scanned and band intensities quantified using the Image Studio 3.1 and LI-COR Odyssey Infrared Imaging system according to manufacturer’s instructions.
Offer S.M., Butterfield G.L., Jerde C.R., Fossum C.C., Wegner N.J, & Diasio R.B. (2014). MicroRNAs miR-27a and miR-27b directly regulate liver dihydropyrimidine dehydrogenase expression through two conserved binding sites. Molecular cancer therapeutics, 13(3), 742-751.
+ Open protocol
+ Expand
4

Western Blot Protein Expression Analysis

Check if the same lab product or an alternative is used in the 5 most similar protocols
Cells were seeded in 6-well plates 24 hours prior to transfection or drug treatment. Samples were harvested 48 hours after drug treatment and 72 hours after transfection. Cells were lysed in RIPA buffer containing protease and phosphatase inhibitor cocktails. Running samples were prepared with 4X Novex NuPAGE LDS sample buffer (Life Technologies) and supplemented with 10% beta-mercaptoethanol. Twenty micrograms of protein were loaded per lane of 4–12% Bis-Tris NuPAGE gels (Life Technologies) and ran in MOPS buffer. Proteins were then transferred to PVDF-FL membrane (Millipore) in Tris-Glycine transfer buffer. Primary antibody incubation was carried out overnight at 4°C on a rocking platform. Secondary antibody incubation was carried out at room temperature the next day. Proteins were detected using the LI-COR Odyssey imaging system (LI-COR Biosciences). Image analysis and protein quantitation were performed with ImageJ (NCI).
Gu S., Hu Z., Ngamcherdtrakul W., Castro D.J., Morry J., Reda M.M., Gray J.W, & Yantasee W. (2016). Therapeutic siRNA for drug-resistant HER2-positive breast cancer. Oncotarget, 7(12), 14727-14741.
+ Open protocol
+ Expand
5

Immunoblotting of Cell Signaling Proteins

Check if the same lab product or an alternative is used in the 5 most similar protocols
Cells were trypsinized and washed in cold PBS before being lysed in 2X Laemmli sample buffer containing 0.5% beta-mercaptoethanol, 1% each of Protease Inhibitor Cocktail, Phosphatase Inhibitor Cocktail 2, and Phosphatase Inhibitor Cocktail 3 (Sigma-Aldrich, P8340, P5726, P0044, respectively). Samples were run on 4-20% Tris-Glycine gels (Life Technologies) and transferred to 0.45 μm PVDF-FL membrane (Millipore). Membranes were blocked in 2% milk solution and probed with the following antibodies: NMIIB (Biolegend, rabbit pAb; 1:1000 cat #909901), GAPDH (Santa Cruz Biotechnology rabbit pAb; 1:5000). EGFR (Cell Signaling Technology, rabbit pAb; 1:1000). Blots were analyzed with LiCor Odyssey CL imaging system.
Thomas D., Thiagarajan P.S., Rai V., Reizes O., Lathia J, & Egelhoff T. (2016). Increased cancer stem cell invasion is mediated by myosin IIB and nuclear translocation. Oncotarget, 7(30), 47586-47592.
+ Open protocol
+ Expand
6

SDS-PAGE and Western Blotting Quantification

Check if the same lab product or an alternative is used in the 5 most similar protocols
Protein samples were resolved on 4–20% SDS-gradient gels (BioRad, Hercules, CA), then transferred to PVDF-FL membrane (Millipore Burlington, MA). Detection and quantification was carried out using a LI-COR Odyssey infrared scanning system using fluorescently labelled secondary antibodies. All western blots were scanned using the Odyssey Clx infrared imaging system from LICOR.
D'Souza R.S., Lim J.Y., Turgut A., Servage K., Zhang J., Orth K., Sosale N.G., Lazzara M.J., Allegood J, & Casanova J.E. (2020). Calcium-stimulated disassembly of focal adhesions mediated by an ORP3/IQSec1 complex. eLife, 9, e54113.
+ Open protocol
+ Expand
7

ATM Signaling Pathway Analysis

Check if the same lab product or an alternative is used in the 5 most similar protocols
Cells were lysed in cell lysis buffer (9803, Cell Signaling) and lysate (10 µg) was separated by SDS-PAGE and analyzed by western blotting. Proteins were transferred to PVDF-FL membrane (Millipore) and probed with antibodies directed against ATM (GRX70103, Genetex), phospho-ATM Ser-1981 (AF-1655, R&D Systems), p53 (GTX70214, Genetex), phospho-p53 Ser-15 (9286, Cell Signaling), Smc1 (4802, Cell Signaling), phospho-Smc1 Ser-957(4805S, Cell Signaling), Kap1 (ab22553, Abcam), phospho-Kap1 Ser-824 (A300-767A, Bethyl Laboratories), Nbs1 (GTX70224, Genetex), phospho-Nbs1 Ser-343 (ab47272, Abcam), Chk2 (GTX70295, Genetex), and phospho-Chk2 Thr-68 (2661S, Cell Signaling) followed by detection with IRdye 800 anti-mouse (Rockland, RL-610-132-121) or Alexa Fluor 680 anti-rabbit (Invitrogen, A21076) secondary antibodies. Western blots were analyzed and quantitated using a Licor Odyssey system.
Lee J.H., Guo Z., Myler L.R., Zheng S, & Paull T.T. (2014). Direct Activation of ATM by Resveratrol under Oxidizing Conditions. PLoS ONE, 9(6), e97969.
+ Open protocol
+ Expand
8

Western Blot Protein Detection Protocol

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, and protein was quantified using BCA assay. Samples were prepared with 4X Novex NuPAGE LDS sample buffer (Life Technologies) and supplemented with 10% beta-mercaptoethanol. Twenty micrograms of protein was loaded per lane of 4%–12% Bis-Tris NuPAGE gels (Life Technologies) and run in MOPS buffer. Following gel electrophoresis, proteins were transferred to a PVDF-FL membrane (Millipore) in tris-glycine transfer buffer. Primary antibody incubation was carried out overnight at 4 °C on a rocking platform. Incubation with IRDye-conjugated secondary antibodies was carried out at room temperature the next day. Proteins were detected using the LI-COR Odyssey imaging system (LI-COR Biosciences). Image analysis and protein quantitation (i.e., band densitometry) was performed with ImageJ (NIH, Washington, DC, USA).
Ngamcherdtrakul W., Sangvanich T., Goodyear S., Reda M., Gu S., Castro D.J., Punnakitikashem P, & Yantasee W. (2019). Lanthanide-Loaded Nanoparticles as Potential Fluorescent and Mass Probes for High-Content Protein Analysis. Bioengineering, 6(1), 23.
+ Open protocol
+ Expand
9

DDR Signaling Activation Assay

Check if the same lab product or an alternative is used in the 5 most similar protocols
For the experiment with IR, cells were irradiated with 10 Gy and incubated for 1hr before harvesting. For the experiments with CPT, H2O2, and arsenite, cells were incubated with media containing 10 μM CPT, 100 μM H2O2, or arsenite for 1hr before harvesting. Cells were lysed in 10× cell lysis buffer (9803, Cell Signaling) and lysate (20 μg) was separated by 8% SDS-PAGE gel and analyzed by western blotting. Proteins were transferred to PVDF-FL membrane (Millipore) and probed with antibodies directed against ATM (sc-135663, Santa Cruz), phospho-ATM Ser-1981 (AF-1655, R&D Systems), KAP1 (ab22553, Abcam), phospho-KAP1 Ser-824 (A300-767A, Bethyl Laboratories), Chk2 (GTX70295, Genetex), and phospho-Chk2 Thr-68 (2661S, Cell Signaling) followed by detection with IRdye 800 anti-mouse (RL-610-132-121, Rockland) or Alexa Fluor 680 anti-rabbit (A21076, Invitrogen) secondary antibodies. Western blots were analyzed and quantitated using a Licor Odyssey system.
Lee J.H., Mand M.R., Kao C.H., Zhou Y., Ryu S.W., Richards A.L., Coon J.J, & Paull T.T. (2018). ATM directs DNA damage responses and proteostasis via genetically separable pathways. Science signaling, 11(512), eaan5598.
+ Open protocol
+ Expand
10

Western Blotting of Phospho-Proteins

Cited 1 time
Check if the same lab product or an alternative is used in the 5 most similar protocols
Cellular lysates were separated by polyacrylamide gel electrophoresis (SDS-PAGE) using 4-15% Criterion gels (Biorad) and transferred to PVDF-FL membrane (Millipore). To analyze site-specific phosphorylation the membranes were blocked for 1 hr at room temperature in a blocking buffer that was 1:1 1x PBS:SEA Block (Thermo Scientific). The PVDF membranes were incubated with primary antibodies against LAT pY132 (Biosource), LAT pY226 (BD Pharmingen), ZAP-70 pY319 (Cell Signaling Technologies), SLP-76 pY128 (BD Pharmingen), PLC-γ1 pY783 (Cell Signaling Technologies), ERK1/ERK2 pT185/pY187 (Thermo Scientific) and GAPDH (Meridian Life Sciences). IRDye 800CW or IRDye 680-conjugated secondary antibodies were diluted in SEA Block as above and incubated with the PVDF membrane for 30 min at room temperature. The membranes were then imaged using the Licor Odyssey. The intensity of the immunoblotting bands was determined using Odyssey’s v3.0 software. Phosphorylated proteins were normalized to GAPDH and the relative amount of phosphorylation to the 2 minute (for LAT pY132, LAT pY226, ZAP-70, SLP-76 and PLC-γ1) or 5 minute (ERK1/ERK2) LAT WT bands were determined as described previously (35 (link)-37 (link)). The data from 4-5 independent replicates were compiled and then plotted using Prism (GraphPad Software).
Tremblay M.M., Ollinger T, & Houtman J.C. (2020). The Membrane Proximal Proline-Rich Region and Correct Order of C-Terminal Tyrosines on the Adaptor Protein LAT Are Required For TCR-Mediated Signaling and Downstream Functions. Cellular signalling, 76, 109790.
+ 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?

Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required

Sign up now

Revolutionizing how scientists
search and build protocols!