Anti tim23

Manufactured by BD

Sourced in United States

Anti-Tim23 is a laboratory equipment product that functions as a tool for research and analysis. It is designed to perform specific tasks related to the study of cellular processes. The core function of Anti-Tim23 is to assist in the investigation of the Tim23 protein complex, which plays a role in the import and sorting of proteins into the mitochondria. The product provides researchers with the means to analyze and understand the mechanisms involved in this cellular process.

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Lab products found in correlation

Lipofectamine 2000, by Thermo Fisher Scientific (3 mentions) P38 inhibitor sb203580, by Merck Group (3 mentions) Anti chchd4, by Proteintech (3 mentions) Anti ha tag clone 3f10, by Roche (3 mentions) Anti β actin and anti alpha tubulin, by Merck Group (3 mentions) Restriction endonuclease, by New England Biolabs (3 mentions) Anti flag, by Merck Group (3 mentions) Anti tom20, by Santa Cruz Biotechnology (2 mentions) Pka inhibitor h89, by Merck Group (2 mentions) Anti pgc 1α, by Santa Cruz Biotechnology (2 mentions) Pi3k inhibitor ly294002, by Merck Group (2 mentions) Anti hsp60, by Enzo Life Sciences (2 mentions) Pkc inhibitor go6983, by Merck Group (2 mentions) Anti cytochrome c, by BD (2 mentions) Bca protein assay kit, by Thermo Fisher Scientific (2 mentions) Alexa fluor 488, by Thermo Fisher Scientific (2 mentions) Micu1, by Abcam (1 mentions) Horseradish peroxidase conjugated secondary antibody, by Jackson ImmunoResearch (1 mentions) Polyvinylidene fluoride membrane, by Bio-Rad (1 mentions) Puromycin, by Thermo Fisher Scientific (1 mentions)

Close spelling variants of this product

TIM23 (32 citations) Mouse anti-TIM23 (7 citations) Anti-TIM23 (611222) (2 citations)

19 protocols using anti tim23

Mitochondrial Protein Expression Analysis

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Brain mitochondria samples were prepared in Laemmli buffer with β-mercaptoethanol, then separated by SDS-PAGE on 12% Tris-acrylamide/bis-acrylamide gels and transferred to polyvinylidene fluoride membranes (BioRad). Membranes were probed overnight with the following primary antibodies: anti-Tim23 (BD Transduction Laboratories), anti-mitochondrial calcium uniporter (MCU, Sigma), anti-voltage dependent anion channel (VDAC, Abcam), anti-mitochondrial calcium uptake 1 (MICU1, Abcam), and anti-cyclophilin D (CypD Mitoscience). Membranes were incubated with horseradish peroxidase-conjugated secondary antibodies (Jackson ImmunoResearch) and detected via chemiluminescence.

Burstein S.R., Kim H.J., Fels J.A., Qian L., Zhang S., Zhou P., Starkov A.A., Iadecola C, & Manfredi G. (2018). Estrogen Receptor Beta Modulates Permeability Transition in Brain Mitochondria. Biochimica et biophysica acta, 1859(6), 423-433.

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Protocols for Lipid Droplet Protein Characterization

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Hoechst 33342, LipidTOX Red, Lipofectamine 2000, OptiPrep and Puromycin were purchased from Invitrogen. Anti-FLAG-M2 beads, DMSO, Sodium oleate, PMSF, Polybrene and Triton X-100 were from Sigma. DSP was from Thermo. cDNAs encoding Rab18, PLIN2, ACSL3 and Sec61β were amplified by PCR from cDNAs of C2C12 cells. Rab18 was cloned into pEGFP-C1. FLAG-tagged Rab18 was cloned into pQCXIP. Rab18-S22N and Rab18-Q67L mutations were constructed by a PCR-based site-directed mutagenesis. Sec61β was cloned into pmCherry-C1, which was modified from pEGFP-C1 by displacing GFP with mCherry. PLIN2 was cloned into pFLAG-CMV4. PLIN2 was cloned into pEGFP-N1, as well as its truncations. FLAG-tagged ACSL3 was cloned into pQCXIP.
Anti-Rab18 (Rabbit, A2812), anti-Tip47 (Rabbit, A6822) and anti-PDI (Rabbit, A0692) antibodies were purchased from ABclonal. Anti-PLIN2 antibody (Rabbit, ab108323) was from Abcam. Anti-LAMP1 antibody (Mouse, 3243s) was from Cell Signaling Technology. Anti-Bip (Mouse, 610979), anti-GM130 (Mouse, 610822) and anti-Tim23 (Mouse, 611223) antibodies were from BD Transduction. Anti-GAPDH antibody (Mouse, MAB374) was from Millipore. Anti-Actin antibody was from Huaxinbio (Mouse, HX1827). Anti-GFP antibody was from Santa Cruz (Mouse, sc-9996). Anti-ACSL3 antibody was from Proteintech (Rabbit, 20710-1-AP).

Deng Y., Zhou C., Mirza A.H., Bamigbade A.T., Xu S., Zhang S., & Liu P. (2020). Rab18 Binds PLIN2 and ACSL3 to Mediate Lipid Droplet Dynamics.

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Characterization of Protein Expression and Apoptosis

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We used the following antibodies: anti-HA tag (clone 3F10) (Roche Applied Science), anti-β-actin and anti-alpha-tubulin (Sigma-Aldrich), anti-GAPDH and anti-Vinculin (Santa Cruz), cleaved PARP, pro-caspase 3 and pro-caspase 8, phospho-H2AX, AIF1, p38, phospho-p38, phospho- Hsp27, phospho-MAPKAP2 (Cell Signaling Technologies, Boston, MA), anti-CHCHD4 (Protein Tech, IL), anti-Tim23 (BD Biosciences, San Diego, CA), cytochrome c (Thermofisher Scientific, MA), Smac (Upstate cell signaling, NY). The peroxidase-conjugated anti-rat, anti-rabbit, anti-mouse and anti-goat antibodies were from Vector Laboratories (Burlingame, CA). Rabbit polyclonal antibodies specific for human CHTM1 and CHCM1/Mic25 were produced via ProSci Inc. (Poway, CA) against full-length recombinant protein. For cell transfections, Polyjet and Lipojet (Signagen Laboratories, Rockville, MD) were used. Expression construct sub-cloning was performed using restriction endonucleases from New England BioLabs (Ipswich, MA). p38 inhibitor-SB203580 was from Sigma-Aldrich (St. Louis, MO) and pan-caspase inhibitor- Z-VAD-FMK was from BD Biosciences (San Jose, CA, USA). Other chemical reagents were obtained from Sigma-Aldrich and Thermo Fisher Scientific.

Babbar M., Huang Y., Curtiss C.M, & Sheikh M.S. (2019). CHTM1 regulates cancer cell sensitivity to metabolic stress via p38-AIF1 pathway. Journal of Experimental & Clinical Cancer Research : CR, 38, 271.

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Antibody Detection of Cellular Proteins

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Mouse monoclonal antibodies used in the study were anti-LC3 (MBL International, MA, USA), anti-translocase of the inner membrane (TIM23 or anti-TIM23, BD Transduction Lab, CA, USA), Parkin (Santa Cruz, TX, USA) and Drp1 (Santa Cruz, TX, USA), while rabbit polyclonal antibodies used were β-actin (Cell Signaling Technology, MA, USA) and cytochrome c (Santa Cruz, TX, USA). Secondary antibodies used were Licor fluorescent antibodies (Licor, NE, USA).

Banerjee K., Munshi S., Xu H., Frank D.E., Chen H.L., Chu C.T., Yang J., Cho S., Kagan V.E., Denton T.T., Tyurina Y.Y., Jiang J.F, & Gibson G.E. (2016). Mild mitochondrial metabolic deficits by α-ketoglutarate dehydrogenase inhibition cause prominent changes in intracellular autophagic signaling: Potential role in the pathobiology of Alzheimer’s disease. Neurochemistry international, 96, 32-45.

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Antibody Characterization for Cell Biology Research

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Antibodies used in this study include antiactin (MAB1501; EMD Millipore), anti–α-adaptin (sc-17771; Santa Cruz Biotechnology, Inc.), anti-CD63 (556019; BD), anti-DNA (61014; Progen), anti-Drp1 (611113; BD), anti-Eps15 (sc-534; Santa Cruz Biotechnology, Inc.), anti-Flag (anti-DDDDK, ab1257; Abcam), anti-Flag (F1804; Sigma-Aldrich), anti-GFP (ab6673; Abcam), anti-GFP (A6455; Invitrogen), anti-LAMP2 (sc-18822; Santa Cruz Biotechnology, Inc.), anti-LBPA (Z-SLBPA; Eschelon), anti-parkin (sc-32282; Santa Cruz Biotechnology, Inc.), anti-PDH E2/E3bp (ab110333; Abcam), anti-PINK1 (6946; Cell Signaling Technology), anti-PMP70 (SAB4200181; Sigma-Aldrich), anti-SDHA (ab14715; Abcam), anti-SNAP29 (ab138500; Abcam), anti-Stx17 (17815–1-AP; ProteinTech), anti-Stx17 (HPA001204; Sigma-Aldrich), anti-TIM23 (611222; BD), anti-TIP47 (Novus Biologicals), anti-TOM20 (sc-11414; Santa Cruz Biotechnology, Inc.), anti-UQCRFS1 (referred to herein as CIII-Rieske, ab14746; Abcam), anti-VAMP7 (sc-166394; Santa Cruz Biotechnology, Inc.), anti-VAMP8 (ab76021; Abcam), anti-VDAC1 (ab14734; Abcam), and anti-Vps41 (ab181078; Abcam). Unless otherwise specified, all reagents were purchased from Sigma-Aldrich. The TIP47 antibody was a gift from P. McPherson (McGill University, Montreal, Quebec, Canada).

McLelland G.L., Lee S.A., McBride H.M, & Fon E.A. (2016). Syntaxin-17 delivers PINK1/parkin-dependent mitochondrial vesicles to the endolysosomal system. The Journal of Cell Biology, 214(3), 275-291.

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Advanced Glycation End-Product Assay

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Chemicals used in the study are Glyoxal (Sigma,128465), MethylGlyoxal (Sigma, M0252), Methyl Methanesulfonate (TCI, M0369), Hydroxyurea (Sigma, H8627), Lysozyme (Sigma, 4403), and nitrocellulose membrane (BIO-RAD,1620112). Antibodies used in the study are anti-CML (Carboxymethyl lysine; MAB3247 R&D Systems, Minneapolis, MN, USA), anti-MAGE (Argpyrimidine specific; ab243074 Abcam), anti-Tim23 (BD Bioscience), anti-HA antibody (GT4810, Sigma), secondary mouse (GE-bioscience).

Susarla G., Kataria P., Kundu A, & D'Silva P. (2023). Saccharomyces cerevisiae DJ-1 paralogs maintain genome integrity through glycation repair of nucleic acids and proteins. eLife, 12, e88875.

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Immunoblot Analysis of Mitochondrial Proteins

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For immunoblots, between 7.5 and 25.0 μg of total protein as assessed by BCA assays (Pierce, Rockford, IL) were loaded onto SDS-PAGE gels and transferred by Transblot semi-dry blotting (BioRad, Hercules, CA). Antibodies used included Anti-SUCLA2 (GeneTex, Irvine, CA) at a dilution of 1:2,500, Anti-ABCB7 (GeneTex) at a dilution of 1:500, Anti-ABCB10 (GeneTex) at a dilution of 1:1,000, Anti-PPOX (generated in house by H.A.D. at U.G.A.) at a dilution of 1:2,000, Anti-FECH (generated in house by H.A.D. at U.G.A.) at a dilution of 1:30,000–50,000, Anti-HSP-60 (Sigma) at a dilution 1:50,000, Anti-Cytochrome C (BD Biosciences, San Jose, CA) at a dilution of 1:5,000, VDAC-1 (Millipore, Billerica, MA) at a dilution of 1:2,000 and Anti-TIM-23 (BD Bioscience) at a dilution of 1:2,000. Secondary antibodies used were Anti-Rabbit IgG (H+L) HRP Conjugate (Promega, Madison, WI) and Anti-mouse IgG (H+L) HRP conjugate at dilutions of 1:30,000–60,000 with SuperSignal West Pico Chemiluminescent substrate (Pierce) and X-ray film or ChemiDoc imaging system (BioRad) for detection.

Medlock A.E., Shiferaw M.T., Marcero J.R., Vashisht A.A., Wohlschlegel J.A., Phillips J.D, & Dailey H.A. (2015). Identification of the Mitochondrial Heme Metabolism Complex. PLoS ONE, 10(8), e0135896.

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Metabolic Modulation of Myoblast Differentiation

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Oligomycin, antimycin, FCCP, rotenone, AICAR, and Luperox (TBHP) were from Sigma. Antibodies against OXPHOS complexes (ab110413) and PGC-1alpha (ab106814) were from Abcam. Anti-GAPDH was from Cell Signaling (2118), anti-laminin from Dako (Z0097), anti-Tim23 (611223) and anti-RalA from BD (610221). Low glucose differentiation medium for immortalized human myoblasts was DMEM-F12 (US Biological) and for primary human skeletal muscle myoblasts (HSMM, Lonza) was DMEM (Gibco) supplemented with glucose (1 g/l) and 2% HS (Gibco). Galactose medium contained DMEM-F12 supplemented with galactose (1 g/l) and 2% HS.

Biesemann N., Ried J.S., Ding-Pfennigdorff D., Dietrich A., Rudolph C., Hahn S., Hennerici W., Asbrand C., Leeuw T, & Strübing C. (2018). High throughput screening of mitochondrial bioenergetics in human differentiated myotubes identifies novel enhancers of muscle performance in aged mice. Scientific Reports, 8, 9408.

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Antibody detection of mitochondrial proteins

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The antibody detecting Magmas-1 and Magmas-2 were raised against the N- and C-terminus unique peptide sequences, MRWGQSASGSSVKFTRLPACP and NSWPQAILLPWPPKTLGLHA, respectively (Abgenex). Human primary antibodies used in this study are previously described (26 (link)). Briefly, anti-Tim23 (BD Biosciences), anti-Tim44 (Sigma), anti-Tim17a (Epitomics), anti-Tim50 (Imgenex Biotech), anti-Magmas, anti-DnaJC19, and anti-DnaJC15 antibodies were raised against the respective J-like and J-domain, whereas anti-Tim17b antibody was generated against the C-terminus peptide CPKDGTPAPGYPSYQ (Imgenex Biotech). In addition, antibodies specific for yeast proteins, anti-Tim44, anti-Pam18, anti-Mge1, and anti-Pam16, were kindly gifted by Prof. Elizabeth A. Craig's laboratory. anti-Tim23 and anti-Tim50 were constructed against the N-terminus (1–98) amino acids and C-terminus (133–476) amino acids, respectively.

Waingankar T.P, & D'Silva P. (2021). Multiple variants of the human presequence translocase motor subunit Magmas govern the mitochondrial import. The Journal of Biological Chemistry, 297(6), 101349.

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Western Blot Analysis of Autophagy Proteins

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Cells were homogenized in RIPA buffer, then separated by SDS-PAGE under reducing conditions and transferred to nitrocellulose membranes (GE Healthcare), blocked with 5% skimmed milk in TBS/0.5% v/v Tween 20 for 1 h, and washed with TBS/Tween 20. Immunoblots were performed using the following primary antibodies: anti-VMP1 (Cell Signaling #12929), anti-ATG5 (Cell Signaling #2630), anti-TIM23 (BD Biosciences #611222) and anti-GAPDH (Enzo ADI-CSA-335). Horseradish peroxidase-conjugated secondary antibodies were purchased from Santa Cruz Biotechnology (goat anti-mouse and goat anti-rabbit). Blots were developed with the enhanced chemiluminescence method following the manufacturer's instructions (Amersham). For western-blot analysis of LC3 (Cell Signaling #2775S) total cell lysates were isolated using RIPA buffer from cells incubated in complete (DMEM) or starvation medium (EBSS) for 4 hours in the presence or absence of 50 μM chloroquine.

Tábara L.C, & Escalante R. (2016). VMP1 Establishes ER-Microdomains that Regulate Membrane Contact Sites and Autophagy. PLoS ONE, 11(11), e0166499.

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