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Immobilon

Immobilon is an AI-powered research platform developed by Millipore Sigma that enables scientists to optimize their experimental protocols.
PubCompare.ai, the flagship tool of Immobilon, leverages advanced natural language processing and machine learning algorithms to analyze data from scientific literature, preprints, and patents.
This empowers researchers to discover the most effective protocols and products, enhancing reproducibility and accuracy in their work.
PubCompare.ai's powerful AI-driven analysis helps users make informed decisions and streamline their research processes.

Most cited protocols related to «Immobilon»

1
Mitochondrial Protein SDD-AGE Protocol
Cited 24 times

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Publication 2011
Bromphenol Blue Buffers Electrophoresis Glycerin Immobilon Mitochondria Proteins Sepharose Tissue, Membrane
2
Western Blot Protein Quantification
Cited 15 times
For gel electrophoresis, tissue samples were adjusted to a concentration of 0.8 μg/μl with sterile water, reducing buffer (Invitrogen), and denaturing buffer (Invitrogen). Samples were then incubated at 70°C for 10 min.
The Novex Mini Cell NuPAGE system (Invitrogen) with 4–12% Bis–Tris gradient polyacrylamide gels (Invitrogen) was used. A 8 μg of denatured protein homogenate was run in each lane. Samples were loaded in duplicate in adjacent lanes, and a molecular weight standard was run on each gel. A lane containing 8 μg of homogenized macaque cortex was also loaded onto each gel to control for interblot variability. Gels were suspended in a bath of NuPAGE MES SDS running buffer (Invitrogen) with 500 μl NuPAGE antioxidant (Invitrogen) during electrophoresis.
Following electrophoresis, proteins were transferred onto Immobilon-FL PVDF membranes (Millipore) using a semi-dry transfer apparatus (Bio-Rad). After electroblot transfer of the protein, membranes were washed twice and incubated with Odyssey Blocking Buffer (Li-Cor Biosciences) for 1 h at room temperature with rocking to block non-specific antibody binding. Membranes were exposed to the primary polyclonal antibody diluted 1:10,000 for actin (Chemicon MAB150R), VCP (Abcam ab11433), and β-tubulin (Upstate 05-661), and 1:20,000 for GAPDH (Sigma G9545) in blocking buffer with 0.1% tween overnight at 4°C with rocking. Next, the membranes were washed three times for 10 min in tris-buffered saline with 0.1% tween (TBST), then rocked for 1 h with anti-mouse IR-Dye 680 or 800 CW secondary antibody (Li-Cor Biosciences) diluted 1:10,000 in blocking buffer with 0.1% tween. Membranes were washed three times for 10 min in TBST then washed five times in high purity water and allowed to dry for 3–5 min before scanning (infrared imaging system; Li-Cor Biosciences). We pre-tested the β-tubulin, actin, GAPDH, and VCP Western blot assays using varying concentrations of protein from a human cortical tissue homogenate sample. These experiments demonstrated that each assay was linear with protein concentrations found in this study (VCP: R = 0.99, P < 0.01; β-tubulin: R = 0.99, P < 0.01; actin: R = 0.95, P = 0.01; GAPDH: R = 0.98, P < 0.01).
Bauer D.E., Haroutunian V., McCullumsmith R.E, & Meador-Woodruff J.H. (2009). Expression of four housekeeping proteins in elderly patients with schizophrenia. Journal of neural transmission (Vienna, Austria : 1996), 116(4), 487-491.
Publication 2009
Actins Antioxidants Bath Biological Assay Bistris Cardiac Arrest Cells Cortex, Cerebral Electrophoresis GAPDH protein, human Gels Homo sapiens Immobilon Immunoglobulins Kidney Cortex Macaca Mus polyacrylamide gels polyvinylidene fluoride Proteins Saline Solution Staphylococcal Protein A Sterility, Reproductive Tissue, Membrane Tissues Tubulin Tweens Western Blot
3
Western Blot Analysis of Neurological Markers
Cited 12 times
Cerebral cortex samples were collected, dissolved and proteins separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis in 10% polyacrylamide gels. Proteins were transferred onto Immobilon nitrocellulose membranes. The membranes were incubated with primary antibodies for 1 hour at room temperature and washed with Tris-buffered saline + 5% Tween-20 (TBST). The following primary antibodies were used: a rabbit polyclonal antibody for anti-LC-3 (1:2000, ab48394; Abcam), a rabbit polyclonal antibody for anti caspase-3 (cell apoptosis marker; 1:300, ab4051; Abcam), and a rabbit polyclonal antibody for anti Beclin-1 (1:3000, ab62557; Abcam). After washing in TBST again, the blots were incubated with horseradish peroxidase-conjugated goat anti-rabbit secondary antibodies (1:1000; Santa Cruz Biotechnology Inc., Santa Cruz, CA, USA) for 1.5 hours at room temperature. Following two rinses and four washes with TBST, the optical density results were quantified by Quantity 4.5 Software (Bio-Rad Laboratories, Hercules, CA, USA) and ImageJ (NIH, Bethesda, MD, USA). GAPDH (1:1000, ab8245; Abcam) was used as internal standards.
Chen J.H., Wu T., Xia W.Y., Shi Z.H., Zhang C.L., Chen L., Chen Q.X, & Wang Y.H. (2020). An early neuroprotective effect of atorvastatin against subarachnoid hemorrhage. Neural Regeneration Research, 15(10), 1947-1954.
Publication 2020
Anti-Antibodies Antibodies Antibodies, Anti-Idiotypic Apoptosis BECN1 protein, human Caspase 3 Cells Cortex, Cerebral GAPDH protein, human Goat Horseradish Peroxidase Immobilon Nitrocellulose polyacrylamide gels Proteins Rabbits Saline Solution SDS-PAGE Tissue, Membrane Tween 20 Vision
4
Western Blotting of p62, Tubulin, and GFP
Cited 10 times
The polyclonal affinity-purified p62 antibody was raised in rabbits using the peptide antigene PRTEDPVTTPRSTQ corresponding to amino acids 297–311 (Genscript). Polyclonal anti-GFP antibodies were raised using standard procedures by immunizing rats with bacterially expressed His-tagged eGFP purified on Ni affinity columns (Sigma). Samples were separated by SDS-PAGE on an 8% acrylamide gel and transferred to Immobilon-P PVDF membrane (Millipore). Membranes were blocked in 3% milk/TBS for 1 h at room temperature and washed three times for 10 min each in TBST (TBS +0.1% Tween-20). Blots were incubated with primary antibodies: rabbit polyclonal anti-p62 [1∶8,000], mouse monoclonal anti-tubulin AA4.3 (DSHB) [1∶200], rat polyclonal anti-GFP [1∶10,000] in 1.5% milk/TBST for 1 h at room temperature, followed by three 10-min washes in TBST. Blots were incubated in AP-conjugated goat anti-rat (Sigma), anti-rabbit or anti-mouse secondary antibody (Millipore) diluted 1∶10,000 in 1.5% milk/TBST for 1 h at room temperature. Blots were washed for 3×10 min in TBST and then incubated with Immobilon Western Chemiluminescent AP Substrate (Millipore), followed by exposure to Super RX film (Fuji).
Pircs K., Nagy P., Varga A., Venkei Z., Erdi B., Hegedus K, & Juhasz G. (2012). Advantages and Limitations of Different p62-Based Assays for Estimating Autophagic Activity in Drosophila. PLoS ONE, 7(8), e44214.
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Publication 2012
Acrylamide Amino Acids Anti-Antibodies Antibodies Antibodies, Anti-Idiotypic Goat Immobilon Immobilon P Immunoglobulins Mice, House Milk, Cow's Oryctolagus cuniculus Peptides polyvinylidene fluoride Rabbits SDS-PAGE Tissue, Membrane Tubulin Tween 20
5
Western Blot Analysis of MEFs
Cited 9 times

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Publication 2013
Antibodies Binding Sites Buffers Cells Cold Temperature Complex, Immune Densitometry Electrophoresis Immobilon Milk, Cow's polyacrylamide gels Powder Proteins Pulse Rate Radioimmunoprecipitation Assay Saline Solution Tissue, Membrane Tweens

Most recents protocols related to «Immobilon»

1
Western Blot Analysis of ER Stress Markers
Total protein was fractionated via 8–12% SDS–PAGE, transferred to an immobilon polyvinylidene difluoride membrane (Amersham Biosciences, Amersham, UK), and hybridized with specific primary antibodies against AGR2 (GeneTex, Hsinchu, Taiwan), ATF6 (GeneTex), p-IRE1α (GeneTex), IRE1α (Cell Signaling Technology, Danvers, MA), p-PERK (GeneTex), PERK (Cell Signaling Technology) and β-actin (GeneTex) overnight at 4 °C. Subsequently, the membrane was probed with the appropriate HRP-conjugated secondary antibody for 1 h at room temperature. Finally, immune complexes were visualized via the chemiluminescence method using an ECL detection kit (Merck, Darmstadt, Germany).
Tsai H.W., Chen Y.L., Wang C.I., Hsieh C., Lin Y.H., Chu P.M., Wu Y.H., Huang Y.C, & Chen C.Y. (2023). Anterior gradient 2 induces resistance to sorafenib via endoplasmic reticulum stress regulation in hepatocellular carcinoma. Cancer Cell International, 23, 42.
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Publication 2023
Actins activating transcription factor 6, human Antibodies Chemiluminescence Complex, Immune ERN1 protein, human Immobilon Immunoglobulins polyvinylidene fluoride Proteins SDS-PAGE Tissue, Membrane
2
Western Blot Analysis of Liver Tissues
Liver tissues were lysed using RIPA lysis buffer and centrifuged at 16,000g for 15 min. LX-2 cells were lysed in RIPA buffer (Cell Signaling Technology, Beverly, MA, USA) containing a cocktail of protease inhibitors (Calbiochem part of Merck KGaA, Darmstadt, Germany) on ice. The supernatants were then obtained after centrifugation at 16,200g for 15 min at 4℃. The protein concentrations of the cell lysates or liver tissues were measured by Bradford assay (Pro-Measure, iNtRON Biotechnology, Seoul, Korea) and the proteins were dissolved in the SDS sample buffer. Western blot analysis was conducted as previously described (Song et al. 2016 (link)). Samples were separated using sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS-PAGE). The separated proteins were transferred to nitrocellulose membranes (GE Healthcare, Madison, WI, USA) and blocked with 5% skim milk. The proteins on the membrane were incubated with the primary antibodies (Supplementary Table 1) overnight. Horseradish peroxidase-conjugated IgG antibodies (Cell Signaling Technology, Beverly, MA, USA) were used as the secondary antibodies. After 2 h incubation with the secondary antibody, immune complexes were detected using the Immobilon Western Chemiluminescent HRP Substrate (Merck Millipore, Billerica, MA, USA).
Kim H.Y., Otgontenger U., Kim J.W., Lee Y.J., Kim S.B., Lim S.C., Kim Y.M, & Kang K.W. (2023). Anti-fibrotic effect of aurocyanide, the active metabolite of auranofin. Archives of Pharmacal Research, 46(3), 149-159.
Publication 2023
Antibodies Biological Assay Buffers Centrifugation Complex, Immune Immobilon Immunoglobulins Introns Liver Membrane Proteins Milk, Cow's Nitrocellulose Protease Inhibitors Proteins Radioimmunoprecipitation Assay SDS-PAGE Tissue, Membrane Tissues Western Blot
3
Hypoxia-Induced Protein Expression Analysis
Following the induction of hypoxia, the cells were quickly collected and lysed using RIPA protein extraction reagent (Epizyme, Inc.; Ipsen) supplemented with protease inhibitor and phosphatase inhibitor cocktails (both Epizyme. Inc.; Ipsen). The concentrations of the protein samples were detected using a BCA Protein Assay kit (Thermo Fisher Scientific, Inc.). Protein extracts (15 µg/lane) were separated by 10% SDS-PAGE and transferred to polyvinylidene fluoride membranes. The membranes were blocked for 25 min at room temperature in protein-free blocking buffer (Epizyme, Inc.; Ipsen) and incubated with primary antibodies at 4°C for 12 h. Antibodies targeting CDK2 (cat. no. A0094), cyclin D1 (cat. no. A19038), AKT1 (cat. no. A20799), phosphorylated-AKT (cat. no. AP1172) and β-actin (cat. no. AC026) were used. All primary antibodies were diluted 1:1,000. All the primary antibodies were acquired from ABclonal Biotech Co., Ltd. Membranes were incubated with Anti-rabbit IgG, HRP-linked Antibody (dilution, 1:5,000; cat. no. 7074; Cell Signaling Technology, Inc.) at room temperature for 60 min. Immobilon Western HRP Substrate (WBKLS0050) was purchased from MilliporeSigma. The membrane was exposed to an autoradiography film and autoradiograms were quantified by densitometry using Quantity One software 4.4.6 (Bio-Rad Laboratories, Inc.).
Sang W., Zhu R., Liu D, & Gong M. (2023). LncRNA SPRY4‑IT1 is upregulated and promotes the proliferation of prostate cancer cells under hypoxia in vitro. Oncology Letters, 25(4), 138.
Publication 2023
Actins AKT1 protein, human anti-IgG Antibodies Autoradiography Biological Assay Buffers Cardiac Arrest CDK2 protein, human Cells Cyclin D1 Densitometry Hypoxia Immobilon Immunoglobulins Phosphoric Monoester Hydrolases polyvinylidene fluoride Protease Inhibitors Proteins Rabbits Radioimmunoprecipitation Assay SDS-PAGE Technique, Dilution Tissue, Membrane
4
Western Blotting of Protein Samples
Protein samples in 1× LDS sample buffer containing 4% BME were separated by size on 4 to 12% Novex NuPage Bis-Tris SPS-PAGE (Invitrogen) gels (2 h, 100 V). Gels were transferred to a Immobilon-FL PVDF Membranes (Millipore) using the Mini Blot Module western blotting system (Invitrogen) for 1 h in transfer buffer (192 mM glycine, 25 mM Tris, 20% methanol [v/v], 4 °C). Membranes were dried for 15 min in the fume hood, rehydrated with a 15 s methanol wash, and submerged in TBST (500 mM NaCl, 20 mM Tris pH 7.4, 0.05% Tween 20 [v/v]). Membranes were blocked in 5% nonfat dry milk (NFDM) in TBST for 1 h on a rocker at RT. Membranes were incubated with primary antibodies diluted (Table S2) in 1% NFDM in TBST with 0.02% (w/v) sodium azide overnight on a rocker at 4 °C. After the primary incubation, membranes were washed in TBST (3×, 5 min, RT) on a rocker. Membranes were incubated on a rocker with fluorophore-conjugated secondary antibodies diluted (Table S2) in 5% NFDM in TBST (1 h, RT). After the secondary incubation, membranes were washed in TBST (3×, 5 min, RT) on a rocker and imaged on LI-COR Odyssey CLx using the Image Studio v5.2 software (LI-COR). Special care was made to ensure that membranes were exposed to the light as little as possible to protect the fluorescent signal of the secondary antibody.
Schmitz J.M., Wolters J.F., Murray N.H., Guerra R.M., Bingman C.A., Hittinger C.T, & Pagliarini D.J. (2023). Aim18p and Aim46p are chalcone isomerase domain–containing mitochondrial hemoproteins in Saccharomyces cerevisiae. The Journal of Biological Chemistry, 299(3), 102981.
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Publication 2023
Antibodies Bistris Buffers Fluorescent Antibody Technique Gels Glycine Immobilon Light Methanol Milk, Cow's polyvinylidene fluoride Proteins Sodium Azide Sodium Chloride Tissue, Membrane Tromethamine Tween 20 Western Blot
5
Cellular Protein Expression Analysis
Pre-cooled H9c2 cells at 4˚C and left ventricular tissue from mice were lysed on ice for 30 min using RIPA Lysis Buffer (MedChemExpress), which contained 20 mmol/l Tris (pH 7.5), 150 mmol/l NaCl, 1% Triton X-100 and 1% Phosphatase Inhibitor Cocktail I and III (MedChemExpress). Next, the cell lysis products were centrifuged for 10 min at 12,700 x g in a 4˚C refrigerated centrifuge and the supernatants were collected. The protein concentration was measured using a BCA protein kit (Thermo Fisher Scientific, Inc.) and the samples were boiled at 100˚C for 5 min. A total of 30 µg cellular protein/lane were electrophoresed using 10% SDS-PAGE, and transferred onto a PVDF membrane. The membrane was blocked for 2 h at room temperature with 5% BSA (Beyotime Institute of Biotechnology), incubated overnight at 4˚C with primary antibodies of GAPDH (dilution, 1:10,000; ab181602; Abcam), cGAS (dilution, 1:1,000; ab252416; Abcam), STING (dilution, 1:1,000; ab288157; Abcam), Bcl-2 (dilution, 1:1,000; ab196495; Abcam), Bax (dilution, 1:1,000; ab32503; Abcam) and cleaved Caspase-3 (dilution, 1:1,000; ab184787; Abcam), washed with TBS containing 0.1% Tween 20, and incubated for 2 h with HRP Anti-Rabbit IgG antibody (dilution, 1:10,000; ab184787; Abcam) at room temperature. The immunoreactivity of the proteins was visualized by chemiluminescence with immobilon western chemilum HRP substrate (MilliporeSigma). Signals were detected and analyzed with ChemiDoc™XRS + with Image Lab™ Software Gel Imaging System (version 2.0; Bio-Rad Laboratories, Inc.).
Li J.K., Song Z.P, & Hou X.Z. (2023). Scutellarin ameliorates ischemia/reperfusion injury‑induced cardiomyocyte apoptosis and cardiac dysfunction via inhibition of the cGAS‑STING pathway. Experimental and Therapeutic Medicine, 25(4), 155.
Publication 2023
anti-IgG Antibodies BCL2 protein, human Buffers Caspase 3 Cells Chemiluminescence Chromogranin A GAPDH protein, human Immobilon Immunoglobulins Left Ventricles Mus Phosphoric Monoester Hydrolases polyvinylidene fluoride Proteins Rabbits Radioimmunoprecipitation Assay SDS-PAGE Sodium Chloride Staphylococcal Protein A Technique, Dilution Tissue, Membrane Tissues Triton X-100 Tromethamine Tween 20

Top products related to «Immobilon»

1
Immobilon western chemiluminescent hrp substrate by Merck Group
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Immobilon Western Chemiluminescent HRP Substrate is a laboratory reagent used for the detection of proteins in Western blot analysis. It is a chemiluminescent substrate that reacts with horseradish peroxidase (HRP) enzyme, enabling the visualization of target proteins on a membrane.
2
Pvdf membrane by Merck Group
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PVDF membranes are a type of laboratory equipment used for a variety of applications. They are made from polyvinylidene fluoride (PVDF), a durable and chemically resistant material. PVDF membranes are known for their high mechanical strength, thermal stability, and resistance to a wide range of chemicals. They are commonly used in various filtration, separation, and analysis processes in scientific and research settings.
3
Immobilon by Merck Group
Sourced in United States, Germany, United Kingdom, Japan, Ireland, France
Immobilon is a line of microporous membrane products used in various laboratory applications. These membranes are designed for effective protein and nucleic acid transfer and immobilization, supporting a wide range of analytical techniques.
4
Immobilon fl pvdf membrane by Merck Group
Sourced in United States, United Kingdom, Germany, Japan, France
Immobilon-FL PVDF membrane is a fluorescence-compatible polyvinylidene difluoride (PVDF) membrane used for protein transfer and detection. It is designed to provide high protein-binding capacity and low background fluorescence for sensitive protein analysis applications.
5
Immobilon pvdf membrane by Merck Group
Sourced in United States, Germany, United Kingdom, China, Ireland
Immobilon PVDF membrane is a laboratory product manufactured by Merck Group. It is a polyvinylidene fluoride (PVDF) membrane material commonly used in various analytical and research applications that require protein or nucleic acid transfer and detection.
6
Immobilon fl by Merck Group
Sourced in United States, Germany, Ireland
Immobilon-FL is a membrane-based system designed for the immobilization and detection of biomolecules, such as proteins and nucleic acids. It provides a versatile platform for various analytical and diagnostic applications.
7
Immobilon western by Merck Group
Sourced in United States, Germany, Japan, United Kingdom, Morocco
Immobilon Western is a laboratory product manufactured by Merck Group. It is a membrane used for protein detection and analysis in Western blotting procedures.
8
Odyssey infrared imaging system by LI COR
Sourced in United States, Germany, United Kingdom, China, Italy, Niger, France, Netherlands, Austria, Australia, Liechtenstein
The Odyssey Infrared Imaging System is a versatile laboratory equipment designed for high-sensitivity detection and quantification of fluorescent and luminescent signals. The system utilizes infrared technology to capture and analyze various molecular targets, such as proteins, nucleic acids, and small molecules, in a range of sample types.
9
Immobilon western hrp substrate by Merck Group
Sourced in United States, Germany, Canada, United Kingdom, China, France, Japan, Czechia
Immobilon Western HRP Substrate is a chemiluminescent substrate used for the detection of horseradish peroxidase (HRP) in Western blotting applications. It is designed to produce a stable, high-intensity signal that can be detected using a chemiluminescence imager or film.
10
Immobilon membrane by Merck Group
Sourced in United States, Germany, United Kingdom, Japan
Immobilon membrane is a microporous membrane made of PVDF (polyvinylidene fluoride) material. It is designed for use in various laboratory applications, including Western blotting, dot blotting, and other immunodetection techniques. The membrane provides a stable and efficient platform for the immobilization of proteins, nucleic acids, and other biomolecules.

More about "Immobilon"

Immobilon is an AI-powered research platform developed by Millipore Sigma that enables scientists to optimize their experimental protocols.
PubCompare.ai, the flagship tool of Immobilon, leverages advanced natural language processing and machine learning algorithms to analyze data from scientific literature, preprints, and patents.
This empowers researchers to discover the most effective protocols and products, enhancing reproducibility and accuracy in their work.
PubCompare.ai's powerful AI-driven analysis helps users make informed decisions and streamline their research processes.
The platform can be used in conjuction with Immobilon Western Chemiluminescent HRP Substrate, PVDF membranes, Immobilon-FL PVDF membrane, Immobilon PVDF membrane, Immobilon-FL, Immobilon Western, and the Odyssey Infrared Imaging System to further optimize experimental workflows and boost research productivity.
Discover how Immobilon's PubCompare.ai can leverage artificial intelligence to take your research to the next level.
Find the best protocols and products by comparing data from literature, pre-prints, and patents.
Enhance reproducibility, accuracy, and efficiency with PubCompare.ai's powerful AI-driven analysis toolset.
Streamline your research process and make more informed decisions with this cutting-edge platform from Millipore Signa.