Immobilion fl pvdf membrane

Manufactured by Merck Group

Sourced in United States

Immobilion-FL PVDF membranes are a type of laboratory equipment used for protein detection and analysis. They are made of polyvinylidene fluoride (PVDF) material and designed for use in fluorescence-based applications.

Automatically generated - may contain errors

Lab products found in correlation

Odyssey blocking buffer, by LI COR (2 mentions) Infrared imaging system, by LI COR (1 mentions) Bradford assay method, by Merck Group (1 mentions) Enhanced chemiluminescence reagent, by Cytiva (1 mentions) X omat film, by Kodak (1 mentions) Irdye 680rd and 800cw secondary antibodies, by LI COR (1 mentions) Horseradish peroxidase conjugated secondary antibody, by Santa Cruz Biotechnology (1 mentions) Nitrocellulose membrane, by Bio-Rad (1 mentions) Phospho eif2α, by Cell Signaling Technology (1 mentions) Total eif2α, by Cell Signaling Technology (1 mentions) Atf4 sc 200, by Santa Cruz Biotechnology (1 mentions) Criterion tris hcl gel, by Bio-Rad (1 mentions) Alpha tubulin, by Merck Group (1 mentions) Gapdh, by Merck Group (1 mentions) Pierce ecl western blotting substrate, by Thermo Fisher Scientific (1 mentions) Bradford assay, by Bio-Rad (1 mentions) Cell extraction buffer, by Thermo Fisher Scientific (1 mentions) P chk2 thr68, by Cell Signaling Technology (1 mentions) Odyssey fc, by LI COR (1 mentions) Na3vo4, by Merck Group (1 mentions)

Close spelling variants of this product

PVDF membranes (20308 citations) PVDF-FL membranes (46 citations) Immobilion (11 citations) Immobilion PVDF membranes (8 citations) PVDF-FL (6 citations) Immobilion FL membrane (5 citations) Immobilion®-FL (2 citations)

5 protocols using immobilion fl pvdf membrane

Western Blot Protein Analysis Protocol

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

Whole cell lysates (WCL) were obtained using lysis buffer (50 mM Tris-HCl, pH 7.4; 1% Nonidet P-40; 0.25% sodium deoxycholate; 150 mM NaCl; 1mM EDTA with protease inhibitors) followed by centrifugation at 15,000 g for 15 minutes at 4°C. Protein concentrations of the lysates were measured by the Bradford assay method (Sigma, MO, USA). Equal amounts of protein were loaded onto SDS-PAGE gel, transferred to nitrocellulose membrane (Bio-Rad, CA, USA) or Immobilion-FL PVDF membrane (EMD Millipore) and incubated overnight with the primary antibody of interest. The horseradish peroxidase-conjugated secondary antibody (1:3000) (Santa Cruz Biotechnologies, TX, USA) or the IRDye 680RD and 800CW secondary antibodies (1:2000) (LI-COR, NE, USA) were used. Proteins of interest were detected using enhanced chemiluminescence reagent (Amersham, IL, USA) and Kodak X-OMAT film or with an infrared imaging system (LI-COR, NE, USA). Actin, GAPDH, α-tubulin and lamin B1 protein levels were used as loading controls.

Tomita K., Kabashima A., Freeman B.L., Bronk S.F., Hirsova P, & Ibrahim S.H. (2017). Mixed Lineage Kinase 3 Mediates the Induction of CXCL10 by a STAT1-Dependent Mechanism during Hepatocyte Lipotoxicity. Journal of cellular biochemistry, 118(10), 3249-3259.

+ Open protocol

+ Expand

Western Blot Analysis of Liver Proteins

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

Liver tissue was homogenized in Tissue Protein Extraction Reagent (T-PER®, Waltham, Massachusetts, USA). Equal amounts of protein were loaded onto Criterion Tris-HCl gels (Bio-Rad Lab, Hercules, CA) and electro-transferred to Immobilion® - FL PVDF membrane (EMD Millipore). The membranes were blocked in either 5% Non-Fat Dry Milk or 5% bovine serum albumin in TBST depending on the primary antibody manufacturer’s recommendation for 1 hour at room temperature. Membranes were then incubated with the primary antibody overnight followed by incubation with horseradish peroxidase-conjugated secondary antibodies for 1 hour. Pierce ECL Western Blotting Substrate (ThermoFisher Scientific, Waltham, Massachusetts, USA) was used to detect the HRP activity of the secondary antibodies and expose to films. Primary antibodies used were: phopsho-PERK (Product # 3179), total PERK (Product # 3192), phospho-eIF2α (9721), and total eIF2α (9722) from Cell Signaling Technology, ATF4 (sc-200) and CHOP (sc-7351) from Santa Cruz Biotechnology, GAPDH (Millipore, Mab374) and alpha-tubulin (Sigma, T9026).

Bandla H., Dasgupta D., Mauer A.S., Nozickova B., Kumar S., Hirsova P., Graham R.P, & Malhi H. (2018). Deletion of endoplasmic reticulum stress-responsive co-chaperone p58IPK protects mice from diet-induced steatohepatitis. Hepatology research : the official journal of the Japan Society of Hepatology, 48(6), 479-494.

+ Open protocol

+ Expand

Tumor Lysate Preparation and Western Blot

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

Tumor lysates were prepared from flash-frozen tumor pieces in RIPA buffer (150mM sodium chloride, 50mM Tris-HCl pH7.4, 2mM EDTA,10m NaF, 10mM Sodium pyrophosphate, 1% sodium deoxycholate, 1% NP-40) supplemented with 1mM sodium orthovanadate, 1mM PMSF, 10mM NaF, 10ug/ul leupeptin and 10ug/ul apotinin. Whole cell lysate was also prepared in RIPA buffer. Protein concentration was quantified by Bradford Assay (Bio-Rad) and 25ug of total protein was resolved by SDS-PAGE, and transferred to Immobilion FL PVDF membranes (Millipore), and block with Odyssey Blocking buffer (Li-COR Biosciences). Primary antibodies were diluted in Odyssey Blocking buffer overnight at 4°C. Secondary fluorophore-conjugated antibodies, IRDye 800 anti-rabbit, IRDye 680 anti-goat and IRDye 680 anti-mouse were incubated for 1hr at room temperature. Immunoblots were scanned with Odyssey CL-X imaging system, and images were processed with Image Studio Lite V4.0.

Xiao B., Zuo D., Hirukawa A., Cardiff R.D., Lamb R., Sonenberg N, & Muller W.J. (2020). Rheb1-independent Activation of mTORC1 in Mammary Tumors Occurs Through Activating Mutations in mTOR. Cell reports, 31(4), 107571.

+ Open protocol

+ Expand

Dose-dependent Chk1/2 and PARP Analysis

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

Prior to lysis, HeLa cells were treated with the IC₅₀ (for analysis of Chk1/2) or IC₈₅ (for analysis of cleaved PARP) concentrations of compounds 2 and 5-8, or DMSO vehicle control for 24 h or a 4, 8, 16, and 24 h time course. Cells were collected by scraping and lysed with cell extraction buffer (Invitrogen) supplemented with protease inhibitor cocktail (Sigma-Aldrich), 50 mM NaF, 200 μM Na₃VO₄ (Sigma-Aldrich), and 1 mM phenylmethylsulfonyl fluoride (PMSF) (Sigma-Aldrich). Protein concentration was determined by a Coomassie Plus assay kit (Life Technologies) and equal amounts of protein were resolved by SDS-PAGE on NuPage Bolt 12% Bis-Tris gels (Life Technologies) and transferred to Immobilion-FL PVDF membranes (Millipore). Membranes were probed with primary antibodies diluted in Odyssey blocking buffer (LI-COR Biosciences, Lincoln, NE, USA) for Chk1, p-Chk1 (Ser345), Chk2, cleaved PARP, and GAPDH at 1:1000 and p-Chk2 (Thr68) at 1:500 (Cell Signaling Technology). IRDye 680 or 800 secondary antibodies were used at 1:20000 (LI-COR Biosciences), and the fluorescence signals were imaged on an Odyssey FC (LI-COR Biosciences).

Jans P.E., Mfuh A.M., Arman H.D., Shaffer C.V., Larionov O.V, & Mooberry S.L. (2017). Cytotoxicity and Mechanism of Action of the Marine-Derived Fungal Metabolite Trichodermamide B and Synthetic Analogues. Journal of natural products, 80(3), 676-683.

+ Open protocol

+ Expand

Western Blot Protein Expression Analysis

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

Cells were placed on ice and the media were aspirated. The cells were washed once with cold phosphate-buffered saline (PBS) and harvested in cold 1 × lysis buffer [50 mM Tris–HCl (pH 7.4), 150 mM NaCl, 1% Triton-100, containing 40 μl/mL complete protease inhibitors (Roche Applied Science)]. Lysates were cleared by centrifugation at 12 000 × g for 10 min at 4°C and the protein content was determined. Equal amounts of protein were electrophoretically separated on 10% SDS/polyacrylamide gels and proteins were transferred onto Immobilion-FL PVDF membranes (Millipore). Membranes were blocked with 5% non-fat milk in PBST for 1 hour at room temperature followed by overnight incubation at 4°C with primary antibodies against Actin (1:40000 MP Biomedicals), α-tubulin (1:4000 Abcam), Lamin B (1:1000 Santa Cruz), Bcl-3 (1:500 Santa Cruz). Primary antibodies were detected with horseradish peroxidase-labelled secondary antibody (1:5000, DAKO). The chemoluminescence was detected with a charge-coupled device camera (Fujifilm).

Saamarthy K., Björner S., Johansson M., Landberg G., Massoumi R., Jirström K, & Masoumi K.C. (2015). Early diagnostic value of Bcl-3 localization in colorectal cancer. BMC Cancer, 15, 341.

+ 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!