Log In Sign up
The largest database of trusted experimental protocols

Anti porin

Manufactured by Thermo Fisher Scientific
Visit Supplier
Sourced in Japan

Anti-Porin is a laboratory equipment product that functions as a protein. It is used in the study and analysis of bacterial cell membranes.

Automatically generated - may contain errors

Lab products found in correlation

Anti ha, by Abmart (3 mentions) Anti flag, by Merck Group (3 mentions) Anti gfp, by Roche (3 mentions) Anti phospho ser thr ampk substrate, by Cell Signaling Technology (2 mentions) Polyvinylidene difluoride membrane, by Merck Group (1 mentions) Anti myc, by Takara Bio (1 mentions) Enhanced chemi luminescence (ecl), by Thermo Fisher Scientific (1 mentions) Bca protein assay kit, by Sangon (1 mentions) Anti myc, by Roche (1 mentions) Anti pep12, by Thermo Fisher Scientific (1 mentions) Anti dpm1, by Thermo Fisher Scientific (1 mentions) Anti histone h4, by Abcam (1 mentions) Anti phospho ampka thr172, by Cell Signaling Technology (1 mentions) Anti actin, by Abmart (1 mentions) Anti atr, by Santa Cruz Biotechnology (1 mentions) Hrp linked anti mouse igg, by Cell Signaling Technology (1 mentions) Ecl prime western blotting detection reagent, by GE Healthcare (1 mentions) Ecl western blotting detection reagent, by GE Healthcare (1 mentions) Horseradish peroxidase conjugated secondary antibody, by Bio-Rad (1 mentions) 13c6 l arginine, by Cambridge Isotopes (1 mentions)

11 protocols using anti porin

1

Plasmid Manipulation and Protein Analysis

Cited 1 time
Check if the same lab product or an alternative is used in the 5 most similar protocols
Standard methods were used for plasmid manipulations and transformation of yeast (Schiestl and Gietz, 1989 (link)). Protein concentrations were determined by the method of Lowry et al (1951) (link). Antibodies against mS37, Atp6, Cor1 and Cox1 were kindly provided by Dr. A. Tzagoloff (Columbia University – New York) anti-Prx1 by Dr. Luis Netto (IB-USP, Sao Paulo/Brazil) (Gomes et al., 2017 (link)); anti-uL-29 by Dr. Martin Ott (Stockholm University - Sweden) (Gruschke et. al., 2010 (link)) anti-uL3, anti-uL16, anti-uL24, and anti-bL31 were described elsewhere (Zeng et al., 2018 (link)); anti-Porin was obtained from Invitrogen (Waltham, MA). All reagents, unless otherwise indicated, were obtained from Sigma-Aldrich (Saint Louis, MO).
Santos B., Zeng R., Jorge S.F., Ferreira-Junior J.R., Barrientos A, & Barros M.H. (2021). Functional Analyses of Mitoribosome 54S Subunit Devoid of Mitochondria-Specific Protein Sequences. Yeast (Chichester, England), 39(3), 208-229.
+ Open protocol
+ Expand
2

Subcellular Protein Localization Analysis

Check if the same lab product or an alternative is used in the 5 most similar protocols
To check the expression or positioning of tagged proteins, yeast cells were cultured in YPG medium, collected at OD 10.0 and analysed by western blot according to the method described by Sui et al.58 (link). The cytoplasmic and mitochondrial fractions of engineered strains were separated using a mitochondria extraction kit (Bioversion). Total protein in full cells were extracted using a yeast protein extraction kit (Sangon Biotech, China). For comparison of the protein expression in cytoplasm and mitochondria engineering strains, total protein in full cells and each subcellular fraction were measured using a BCA protein assay kit (Sangon Biotech, China) and 4 μg of protein was loaded in each lane. The protein samples were fractionated on 12% SDS–polyacrylamide gel electrophoresis gel and transferred to polyvinylidene difluoride membranes (Millipore). The membranes were incubated with the indicated primary and secondary antibodies (anti-myc, Clontech; anti-porin, Invitrogen; anti-flag, MBL). Proteins were ultimately visualized by enhanced chemiluminescence and autoradiography (ECL; Thermo Scientific, Waltham, MA, UK).
Lv X., Wang F., Zhou P., Ye L., Xie W., Xu H, & Yu H. (2016). Dual regulation of cytoplasmic and mitochondrial acetyl-CoA utilization for improved isoprene production in Saccharomyces cerevisiae. Nature Communications, 7, 12851.
+ Open protocol
+ Expand
3

Antibody Dilution Protocol for Western Blotting

Check if the same lab product or an alternative is used in the 5 most similar protocols
Antibodies were purchased and used at the indicated dilutions in this study as follows: anti-HA (Abmart, M20003L; 1:3,000), anti-FLAG (Sigma, F1804; 1:2,500), anti-GFP (Roche, 11814460001; 1:2,500), anti-Pgk1 (Nordic Immunology, NE130/7S; 1:10,000), anti-Porin (Invitrogen, 459500, 1:10,000); Anti-Atg17 antibody (gifted from Prof. Yoshinori Ohsumi, Tokyo Institute of Technology, Tokyo); goat anti-mouse IgG1, human ads-HRP (SouthernBiotech, 1070-05; 1:10,000), goat anti-rabbit, human ads-HRP (SouthernBiotech, 4010-05; 1:10,000).
Yao W., Li Y., Chen Y., Chen Y., Zhao P., Zhang Y., Jiang Q., Feng Y., Yang F., Wu C., Zhong H., Zhou Y., Sun Q., Zhang L., Liu W, & Yi C. (2022). Mec1 regulates PAS recruitment of Atg13 via direct binding with Atg13 during glucose starvation-induced autophagy. Proceedings of the National Academy of Sciences of the United States of America, 120(1), e2215126120.
+ Open protocol
+ Expand
4

Antibody Validation and Dilution Protocol

Check if the same lab product or an alternative is used in the 5 most similar protocols
Antibodies were purchased as follows and used at the indicated dilutions: anti-GFP (Roche, 11814460001, 1:2500), anti-MYC (Roche, 11667149001, 1:2500), anti-HA (Abmart, M20003L, 1:2500), anti-FLAG (Sigma, F1804, 1:2500), anti-phospho-AMPKa (Thr172) (Cell Signaling Technology, 2535S, 1:1000), anti-Phospho-(Ser/Thr) AMPK Substrate (Cell Signaling Technology, 5759S, 1: 1000), anti-PGK1 (Nordic Immunology, NE130/7S, 1:10000), anti-Dpm1 (Invitrogen, A6429, 1:2000), anti-ALP (Invitrogen, A6458, 1:2000), anti-Pep12 (Invitrogen, A21273, 1:2000), anti-Porin (Invitrogen, A6449, 1:20000), anti-Histone H4 (Abcam, Ab10158, 1:2500), anti-LC3 (MBL, PM036, 1:1000), anti-ATR (Santa Cruz, SC-1887, 1:250), anti-Actin (Abmart, P30002, 1:5000). Goat Anti-Mouse IgG1, Human ads-HRP (SouthernBiotech, 1070-05, 1:10000) and Goat Anti-Rabbit IgG, Human ads-HRP (SouthernBiotech, 4010-05, 1:10000).
Yi C., Tong J., Lu P., Wang Y., Zhang J., Sun C., Yuan K., Xue R., Zou B., Li N., Xiao S., Dai C., Huang Y., Xu L., Li L., Chen S., Miao D., Deng H., Li H, & Yu L. (2017). Formation of a Snf1-Mec1-Atg1 Module on Mitochondria Governs Energy Deprivation-Induced Autophagy by Regulating Mitochondrial Respiration. Developmental cell, 41(1).
+ Open protocol
+ Expand
5

Western Blot Detection of Cellular Proteins

Check if the same lab product or an alternative is used in the 5 most similar protocols
Proteins were separated by SDS-PAGE. Routinely, proteins were transferred to nitrocellulose membranes (Amersham Biosciences Protran Premium, GE Healthcare) and incubated with primary antibodies (anti-porin (Thermo Fisher Scientific—16G9E6BC4), anti-Pgk1 (Nordic Immunology—NE130/7S), and anti-Tsa1 antibody (from F.G.’s lab). After washing the membranes, the specific secondary antibodies (anti-rabbit IgG, HRP-linked (Cell Signaling Technology) and anti-mouse IgG HRP-linked (Cell Signaling Technology)) were added. The immunoblots were developed using the ECL prime Western blotting detection reagent (GE Healthcare). The normalized band densitometry measurements were performed with the aid of the ImageJ software [17 (link)].
Condeles A.L., Gomes F., de Oliveira M.A., Soares Netto L.E, & Toledo Junior J.C. (2020). Thiol Peroxidases as Major Regulators of Intracellular Levels of Peroxynitrite in Live Saccharomyces cerevisiae Cells. Antioxidants, 9(5), 434.
+ Open protocol
+ Expand
6

Mitochondrial Protein Quantification via Western Blot

Check if the same lab product or an alternative is used in the 5 most similar protocols
Intact mitochondria were isolated using a previously described method (24 (link)). Steady-state levels of mitochondrial proteins were resolved on SDS-PAGE, transferred to nitrocellulose membrane, probed using the indicated primary antibodies and visualized using Amersham enhanced chemiluminescence (ECL) Western Blotting Detection Reagent (GE life sciences, catalog #RPN2106) with horseradish peroxidase-conjugated secondary antibodies (BioRad, catalog #1662408EDU). We used previously described polyclonal rabbit antibodies for immunodetection of Sdh1 and Sdh2 (25 (link)). Anti-porin was purchased from ThermoFisher Scientific (catalog # 459500).
Bannon A.E., Kent J., Forquer I., Town A., Klug L.R., McCann K., Beadling C., Harismendy O., Sicklick J.K., Corless C., Shinde U, & Heinrich M.C. (2017). Biochemical, Molecular, and Clinical Characterization of Succinate Dehydrogenase Subunit A Variants of Unknown Significance. Clinical cancer research : an official journal of the American Association for Cancer Research, 23(21), 6733-6743.
+ Open protocol
+ Expand
7

Protein Detection and Quantification

Check if the same lab product or an alternative is used in the 5 most similar protocols
A mouse monoclonal anti-FLAG antibody, horseradish-peroxidase couple anti-rabbit and anti-mouse antibodies were purchased from Santa Cruz Biotech, TX. Anti-tubulin, anti-Vps10p and anti-PMA1 antibodies were obtained from Abcam, MA. Anti-ALP, anti-porin and anti-dpm1p antibodies were purchased from Thermo Fisher Scientific, MA. D-erythro-sphingosine, D-ribo-phytosphingosine and C16:0-phytoceramide were obtained from Avanti Polar Lipids, AL. L-Lysine (13C6) and L- Arginine (13C6) were obtained from Cambridge Isotope laboratories Inc, MA.
Yi J.K., Xu R., Jeong E., Mileva I., Truman J.P., Lin C.L., Wang K., Snider J., Wen S., Obeid L.M., Hannun Y.A, & Mao C. (2016). Aging-related elevation of sphingoid bases shortens yeast chronological life span by compromising mitochondrial function. Oncotarget, 7(16), 21124-21144.
+ Open protocol
+ Expand
8

Immunoblotting Antibody Reagents and Dilutions

Check if the same lab product or an alternative is used in the 5 most similar protocols
The company, product number and dilution ratio of antibody in this study are as follows: anti-GFP (Roche, 11814460001, 1:2500), anti-FLAG (Sigma, F1804, 1:2500), anti-HA (Abmart, M20003L, 1:3000), anti-phospho-AMPKα(Thr172) (Cell Signaling Technology, 2535S, 1:1000), anti-Phospho-(Ser/Thr) AMPK Substrate (Cell Signaling Technology, 5759, 1:1000), anti-Porin (Thermo Fisher Scientific, A6449, 1:10000), anti-PGK1 (Nordic Immunology, NE130/7S, 1:10000), Goat anti-Mouse IgG1, Human ads-HRP (SouthernBiotech, 1070-05, 1:10000), Goat anti-Rabbit, Human ads-HRP (SouthernBiotech, 4010-05, 1:10000).
Wu C., Yao W., Kai W., Liu W., Wang W., Li S., Chen Y., Wu X., Wang L., Li Y., Tong J., Qian J., Zhang L., Hong Z, & Yi C. (2020). Mitochondrial Fusion Machinery Specifically Involved in Energy Deprivation-Induced Autophagy. Frontiers in Cell and Developmental Biology, 8, 221.
+ Open protocol
+ Expand
9

Quantitative Immunoblotting Analysis of Mitochondrial Complexes

Check if the same lab product or an alternative is used in the 5 most similar protocols
Electrophoresed gels were blotted onto polyvinylidene fluoride (PVDF) membranes using an iBlot transfer system (Invitrogen) according to the manufacturer’s instructions. Blotted PVDF membranes were blocked at room temperature for 30 min. Primary antibody probing was performed at room temperature for 90 min. Secondary antibody probing was performed with chromogenic antibody detection kit (WesternBreeze; Invitrogen) according to the manufacturer’s instructions. Primary antibodies used for SDS-PAGE were as follows: 0.5 μg/mL anti-porin (Molecular Probes), 2.5 μg/mL anti-MT-CO1 (Molecular Probes), 2.5 μg/mL anti-MT-CO2 (Molecular Probes), 2.5 μg/mL anti-COX4 (Molecular Probes), 2.5 μg/mL anti-COX5B (Molecular Probes). Primary antibodies used for BN-PAGE were as follows: 0.5 μg/mL anti-NDUFA9 for CI (Molecular Probes), 0.5 μg/mL anti-SDHA for CII (Molecular Probes), 0.5 μg/mL anti-UQCRC2 for CIII (Molecular Probes), 2.5 μg/mL anti-MT-CO1 for CIV (Molecular Probes), 0.5 μg/mL anti-ATP5B for CV (Molecular Probes).
Hatakeyama H., Katayama A., Komaki H., Nishino I, & Goto Y.I. (2015). Molecular pathomechanisms and cell-type-specific disease phenotypes of MELAS caused by mutant mitochondrial tRNATrp. Acta Neuropathologica Communications, 3, 52.
+ Open protocol
+ Expand
10

Immunofluorescence Microscopy of Yeast Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols
Cells were fixed with 3.5% formaldehyde in the culture medium for 1 h at room temperature. After washing with SP buffer (0.8 M sorbitol, 25 mM potassium phosphate buffer, pH 7.5), cells were pretreated with 0.4 M β-mercaptoethanol for 30 min. After washing with SP buffer, cells were treated with Zymolyase 20T in SP buffer for 30 min to digest the cell wall. Immunofluorescence microscopy was performed according to a method described previously (Kilmartin and Adams, 1984 (link)). Primary antibodies used were anti-Ald4p antibody (rabbit polyclonal, 1:50 dilution, this study) and anti-porin (mouse, monoclonal, Molecular Probe Inc., 1:20 dilution). Alexa 546 goat anti-rabbit IgG (Molecular Probe) and Alexa Fluor 488 goat anti-mouse IgG (Molecular Probe) were used as secondary antibodies at 1:500 dilution. Immunostained cells were double-stained with DAPI (1 µg/ml). Preimmune serum was used in the same method as a control.
Misonou Y., Kikuchi M., Sato H., Inai T., Kuroiwa T., Tanaka K, & Miyakawa I. (2014). Aldehyde dehydrogenase, Ald4p, is a major component of mitochondrial fluorescent inclusion bodies in the yeast Saccharomyces cerevisiae. Biology Open, 3(5), 387-396.
+ 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!