Immunobilon p membrane

Manufactured by Merck Group

Sourced in United States

Immunobilon-P membranes are a type of lab equipment produced by Merck Group. They are designed for use in various immunological and biochemical applications. The core function of these membranes is to facilitate the transfer and immobilization of biomolecules, such as proteins, nucleic acids, or antibodies, for further analysis and processing.

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Lab products found in correlation

Supersignal west femto maximum sensitivity substrate kit, by Thermo Fisher Scientific (4 mentions) Mouse antibody against β actin, by Abcam (2 mentions) Protease inhibitor cocktail, by Merck Group (2 mentions) Ripa lysis buffer, by Thermo Fisher Scientific (2 mentions) Lysis buffer, by Cell Signaling Technology (1 mentions) Sirt6, by Merck Group (1 mentions) Sirt2, by Merck Group (1 mentions) Precision plus protein standard, by Bio-Rad (1 mentions) Ab24607, by Abcam (1 mentions) Enhanced chemi luminescence (ecl), by Thermo Fisher Scientific (1 mentions) Dc protein assay, by Bio-Rad (1 mentions) Ab8224, by Abcam (1 mentions) Nrf2 primary antibody, by Cell Signaling Technology (1 mentions) P chk1, by Cell Signaling Technology (1 mentions) P chk2, by Cell Signaling Technology (1 mentions) P h2ax, by Cell Signaling Technology (1 mentions) β actin, by Abcam (1 mentions) Anti β actin ab, by Merck Group (1 mentions) Amersham imager 680, by GE Healthcare (1 mentions) Enhanced chemiluminescence detection system, by PerkinElmer (1 mentions)

Close spelling variants of this product

Immunobilon-P transfer membranes Immunobilon-P PES membrane

7 protocols using immunobilon p membrane

Western Blot Analysis of Histone Modifications

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Embryos or cells were lysed by sonication in lysis buffer (Cell Signaling Technology, #9803) with a Protease inhibitor cocktail (Sigma) (Wang et al. 2015e (link), Yang et al. 2013 (link), Gu et al. 2015a , Wang et al. 2015a (link), Wang et al. 2015d (link)). Equal amounts of protein from embryos or cells were resolved by SDS-PAGE gel electrophoresis and transferred onto Immunobilon-P membranes (Millipore, Billerica, MA). 2 μg Precision Plus Protein Standards (Bio-Rad, Hercules, CA) were loaded into one lane of the gel. Membranes were incubated in 5% nonfat milk for 45 minutes and then incubated for 18 hours at 4°C with the following primary antibodies at dilutions of 1:1000 to 1:2000 in 5% nonfat milk: SIRT2, SIRT6 from Sigma-Aldrich (St. Louis, MO), acetylated H3K56, H4K16, H4K9, and H3K27 from Cell Signal Technology (Danvers, MA). Membranes were then exposed to goat anti-rabbit or anti-mouse secondary antibodies. To ensure that equivalent amounts of protein were loaded among samples, membranes were stripped and probed with a mouse antibody against β-actin (Abcam, Cambridge, MA). Signals were detected using the SuperSignal West Femto Maximum Sensitivity Substrate kit (Thermo Scientific, Rockford, IL). All experiments were repeated three times.

Yu J., Wu Y, & Yang P. (2016). High glucose-induced oxidative stress represses sirtuin deacetylase expression and increases histone acetylation leading to neural tube defects. Journal of neurochemistry, 137(3), 371-383.

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Western Blot Analysis of Protein Expression

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Cells were lysed in TNT lysis buffer (50 mM Tris-HCl (pH 7.4), 150 mM NaCl, 1% Triton X-100, 1 mM EDTA, 1 mM EGTA, 50 mM NaF, 1 mM Na₃VO₄, 1 mM PMSF, and protease inhibitor cocktail (Sigma)) and protein amounts from cell lysates were measured using the DC protein assay (Bio-Rad). Equivalent amounts of proteins were subjected to SDS-PAGE, transferred to Immunobilon-P membranes (Millipore), immunoblotted with anti-GFP antibody (JL-8, 632381, Takara, 1:1000), anti-Cre recombinase antibody (ab24607, Abcam, 1:1000), anti-Hsp90 antibody (675402, Biolegend, 1:1000), and developed with ECL (Thermo fisher scientific). Immunoblotting with anti-HSP90 antibody was used as a loading control. Alternatively, dissected femurs and soft tissues were homogenized in RIPA lysis buffer (89900, Thermo fisher scientific) and tissue lysates were subjected to immunoblotting analysis.

Yang Y.S., Xie J., Wang D., Kim J.M., Tai P.W., Gravallese E., Gao G, & Shim J.H. (2019). Bone-targeting AAV-mediated silencing of Schnurri-3 prevents bone loss in osteoporosis. Nature Communications, 10, 2958.

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Western Blot Protein Analysis

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Equal amounts of protein (30 μg) from cultured cells or embryos were loaded on a sodium dodecyl sulfate–polyacrylamide (SDS-PAGE) gel and transferred onto Immunobilon-P membranes (Millipore, Billerica, MA) after electrophoresis. Membranes were blocked in 5% nonfat milk for 1 hour, and then incubated overnight at 4°C with the following primary antibodies diluted 1:1000 in 5% nonfat milk: FoxO3a (no. 2497), phosphor-FoxO3a (no. 9464), and LC3 (no. 2775) antibodies (Cell Signaling Technology, Danvers, MA). Membranes were then exposed to horseradish peroxidase (HPR)-conjugated goat antirabbit secondary antibodies. To calibrate protein amounts, membranes were stripped and probed with a mouse anti-β-actin antibody, 1:10,000 (ab8224; Abcam, Cambridge, United Kingdom). Signals were detected using the Super Signal West Femto maximum sensitivity substrate kit (Thermo Scientific, Waltham, MA). Quantification of blots was performed using VisionWorksLS software (UVP Company, Upland, CA).

Xu C., Chen X., Reece E.A., Lu W, & Yang P. (2018). The increased activity of a transcription factor inhibits autophagy in diabetic embryopathy. American journal of obstetrics and gynecology, 220(1), 108.e1-108.e12.

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Nrf2 Protein Expression Analysis

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Equal amounts of protein (30 or 50 μg) from cultured cells and embryos were resolved by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and transferred onto Immunobilon-P membranes (Millipore, Billerica, MA). Membranes were incubated in 5% nonfat milk for 1 hour, and then incubated for 18 hours at 4°C with Nrf2 primary antibodies (Cell Signaling Technology, Danvers, MA) at dilutions of 1:1000 in 5% nonfat milk. Membranes were then exposed to goat anti-rabbit secondary antibodies. To ensure that equivalent amounts of protein were loaded, membranes were stripped and probed with a mouse antibody against β-actin (1:5000; Abcam, Cambridge, UK). Signals were detected using the SuperSignal West Femto Maximum Sensitivity Substrate kit (Thermo Scientific, Waltham, MA). Quantification of blots was performed using VisionWorksLS software (UVP Company, Upland, CA). All experiments were repeated in triplicate.

Zhao Y., Dong D., Reece E.A., Wang A.R, & Yang P. (2017). Oxidative stress-induced miR-27a targets the redox gene Nrf2 in diabetic embryopathy. American journal of obstetrics and gynecology, 218(1), 136.e1-136.e10.

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Western Blot Analysis of DNA Damage Markers

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Equal amounts of protein from embryos or cells were resolved by the SDS-PAGE gel electrophoresis and transferred onto Immunobilon-P membranes (Millipore, Billerica, MA). Membranes were incubated in 5% nonfat milk for 45 minutes and then were incubated for 18 hours at 4°C with the following primary antibodies at dilutions of 1:1000 in 5% nonfat milk: p-Chk1, Chk1, p-Chk2, Chk2, p53, p-H2A.X, H2A.X and SOD1(Cell Signaling Technology). Membranes were then exposed to goat anti-rabbit or anti-mouse secondary antibodies. To confirm that equivalent amounts of protein were loaded among samples, membranes were stripped and probed with a mouse antibody against β-actin (Abcam). Signals were detected using the SuperSignal West Femto Maximum Sensitivity Substrate kit (Thermo Scientific). All experiments were repeated three times with the use of independently prepared tissue lysates.

Dong D., Yu J., Wu Y., Fu N., Villela N.A, & Yang P. (2015). Maternal diabetes triggers DNA damage and DNA damage response in neurulation stage embryos through oxidative stress. Biochemical and biophysical research communications, 467(2), 407-412.

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Immunoblotting of Alpha-Synuclein Protein

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Protein samples were loaded onto 10% SDS-polyacrylamide gels. Following electrophoresis, proteins were transferred to an immunobilon-P membrane (Millipore, Bedford, MA, USA).
Membranes were blocked for 1 hour with 5% nonfat milk in Tris-buffered saline containing 0.05% Tween 20 (TBST). Membranes were probed with the specified primary antibodies diluted in 5% nonfat milk in TBST. Anti α-Syn ab: α-Syn#10¹⁹, a human α-Syn specific; and anti α-Syn#3, recognizes mouse and human α-Syn, anti β-actin ab (Sigma-Aldrich). After washes with TBST, the blots were incubated with secondary antibodies conjugated to horseradish-peroxidase. Immunoreactive bands were detected by the enhanced chemiluminescence reagent (ECL) (Pierce, CT, USA).

Abd-Elhadi S., Honig A., Simhi-Haham D., Schechter M., Linetsky E., Ben-Hur T, & Sharon R. (2015). Total and Proteinase K-Resistant α-Synuclein Levels in Erythrocytes, Determined by their Ability to Bind Phospholipids, Associate with Parkinson’s Disease. Scientific Reports, 5, 11120.

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Western Blot Analysis of Protein Phosphorylation

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Cell lysates were separated by SDS-PAGE using 8~10% gels. The separated proteins were transferred onto an Immunobilon-P membrane (Millipore, Billerica, MA). Blots were blocked with 3% BSA in PBS containing 0.05% Tween 20. The membrane was first probed with primary antibodies. After being washed, the blots were incubated with anti-mouse or rabbit IgG conjugated with HRP. After washing, bound conjugates on the membrane were visualized with an Enhanced Chemiluminescence detection system (PerkinElmer Life Sciences, Waltham, MA) or ImmunoStar LD (Wako Pure Chemical, Osaka, Japan). Chemiluminescence was detected and analyzed by Amersham Imager 680 (GE Healthcare UK Ltd, Buckinghamshire, UK). Chemiluminescence for PY20 was detected by Super RX fuji medical X-ray film (Fuji film). Signal intensity was analyzed by Image J software [37 (link)].

Ohmi Y., Kambe M., Ohkawa Y., Hamamura K., Tajima O., Takeuchi R., Furukawa K, & Furukawa K. (2018). Differential roles of gangliosides in malignant properties of melanomas. PLoS ONE, 13(11), e0206881.

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