P62 gp62 c

Manufactured by Progen Biotechnik

The P62 (GP62-C) is a high-performance laboratory centrifuge designed for a wide range of applications. It features a maximum speed of 6,000 rpm and a maximum relative centrifugal force of 4,400 x g. The centrifuge is equipped with a microprocessor-controlled system that ensures precise speed and time settings. The P62 (GP62-C) can accommodate various rotor types to accommodate different sample volumes and tube sizes.

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Lab products found in correlation

Neun abn78, by Merck Group (1 mentions) Goldenhance, by Nanoprobes (1 mentions) Ubiquitin, by Agilent Technologies (1 mentions) Ab57602, by Abcam (1 mentions) Ab110333, by Abcam (1 mentions) Ab56889, by Abcam (1 mentions) Keap1, by Proteintech (1 mentions) Tom20, by Santa Cruz Biotechnology (1 mentions) Alexa 488 anti rabbit igg, by Thermo Fisher Scientific (1 mentions) Anti mouse igg alexa 488, by Thermo Fisher Scientific (1 mentions) Alexa 568 anti mouse igg, by Thermo Fisher Scientific (1 mentions) Alexa 647 anti mouse igg, by Thermo Fisher Scientific (1 mentions) Tim23, by BD (1 mentions) Tim 44, by BD (1 mentions) Hsp60, by Cell Signaling Technology (1 mentions) Gapdh, by Thermo Fisher Scientific (1 mentions) E cadherin, by Cell Signaling Technology (1 mentions) Lamin a c 2032, by Cell Signaling Technology (1 mentions) Lamp1, by Cell Signaling Technology (1 mentions) Gabarapl1, by Cell Signaling Technology (1 mentions)

5 protocols using p62 gp62 c

Immunohistochemical Analysis of Mouse Brain

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Mouse brains were fixed and dissected as described above. Thick sections were washed with HEPES buffered saline (HBS) pH 7.4 before being permeabilised in HBS containing 0.05 % Triton x-100 and 10 % BSA. Tissue was incubated with primary antibodies overnight at 4 °C in HBS with 0.005 % Triton x-100 and 1 % BSA. Primary antibodies used were as follows: LAMP1 (1D4B, University of Iowa Hybridoma Bank), LAMP2 (Abl-93, University of Iowa Hybridoma Bank), p62 (GP62-C, PROGEN), β-III tubulin (D71G9, cell signalling), NeuN (ABN78, Millipore), GFAP (Z0334, Dako) and Iba1 (ab5076, Abcam). Tissue was then washed before being incubated with either anti-mouse or anti-rabbit nanogold secondaries (Nanoprobes, USA). The tissue was washed before being refixed with 2 % PFA/2.5 % glutaraldehyde in 0.1 M cacodylate buffer. Nanogold particles were then enhanced using Goldenhance (Nanoprobes, USA). Tissue was then post-fixed with 1 % osmium tetroxide and processed for resin embedding as described above.

Clayton E.L., Mizielinska S., Edgar J.R., Nielsen T.T., Marshall S., Norona F.E., Robbins M., Damirji H., Holm I.E., Johannsen P., Nielsen J.E., Asante E.A., Collinge J, & Isaacs A.M. (2015). Frontotemporal dementia caused by CHMP2B mutation is characterised by neuronal lysosomal storage pathology. Acta Neuropathologica, 130(4), 511-523.

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Mitochondrial Dynamics and Protein Quantification

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The following primary antibodies were used: Drp1 (611113; BD Biosciences), Opa1 (612607; BD Biosciences), PDH (ab110333; Abcam), GAPDH (MA5-15738; Thermo), Tim44 (612582; BD Transduction Laboratories), Tim23 (611223; BD Transduction Laboratories), Tom20 (sc-11415; Santa Cruz Biotechnology), mitofusin 1 (ab57602; Abcam), mitofusin 2 (ab56889; Abcam), HA (600-401-384; Rockland), VDAC (4866; Cell Signaling Technology), HSP60 (12165; Cell Signaling Technology), E-cadherin (14472; Cell Signaling Technology), p62 (GP-62C; Progen), ubiquitin (z0458; DAKO), phospho-p62 (serine 351) (PM074; MBL International), LC3 (PM036; MBL) and Keap1 (10503-2-AP; Proteintech). The following secondary antibodies were purchased from Invitrogen: Alexa 488 anti-Rabbit IgG (A21206), Alexa 488 anti-Mouse IgG (A21202), Alexa 488 anti-guinia pig IgG (A11073), Alexa 568 anti-mouse IgG (A10037), Alexa 647 anti-mouse IgG (A31571) and Alexa 647 anti-guinia pig IgG (A21450).

Yamada T., Murata D., Adachi Y., Itoh K., Kameoka S., Igarashi A., Kato T., Araki Y., Huganir R.L., Dawson T.M., Yanagawa T., Okamoto K., Iijima M, & Sesaki H. (2018). Mitochondrial Stasis Reveals p62-mediated Ubiquitination in Parkin-independent Mitophagy and Mitigates Nonalcoholic Fatty Liver Disease. Cell metabolism, 28(4), 588-604.e5.

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Comprehensive Antibody Mapping for Cell Biology

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Primary antibodies for Grp78 (610978) and CPE (610758) were from BD (Transduction); GFAP (Z0334) and Ub (Z0458) from DAKO; Grp94 (SPA-850), PDI (SPA-891) and Hsp40 (Hdj1) (SPA-400) from Enzo; KDEL (PM059) and p62 (PM045) from MBL; APP (MAB348), ChAT (AB144P), Sec61β (07-205) and Ub (MAB1510) from MILLIPORE; HSP 60 (K-19) (sc-1722), NF-YA (sc-10779), Hsp90α (sc-8262) and Hsp70 (W27) (sc-24) from Santa Cruz; GFP (ab13970) and LacZ (ab9361) from abcam; Iba1 (019-19741) from WAKO; LacZ (200-4136) from Rockland; p62 (GP62-C) from PROGEN; Hsp40 (Hdj2) (MS-255-P) form Neomarkers; Lamin A/C (2032) from Cell Signaling.

Yamanaka T., Tosaki A., Miyazaki H., Kurosawa M., Koike M., Uchiyama Y., Maity S.N., Misawa H., Takahashi R., Shimogori T., Hattori N, & Nukina N. (2016). Differential roles of NF-Y transcription factor in ER chaperone expression and neuronal maintenance in the CNS. Scientific Reports, 6, 34575.

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Antibody Panel for Autophagy Signaling

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Antibodies used in this study were as follows: ATG16L1 (8089, human), phospho-ATG14 S29 (92340), ATG14 (96752), phospho-Beclin S30 (54101), FIP200 (12436), FLCN (3697), GABARAPL1 (26632), GABARAPL2 (14256), glyceraldehyde-3-phosphate dehydrogenase (GAPDH) [5174, 1:10,000 for WB (Western blot)], DYKDDDDK tag (14793), HA tag (3724), myc tag (2278), LC3A/B (12741), LC3B (3868), LAMTOR1 (8975), LAMP1 [15665, 1:1000 for immunofluorescence (IF)], NFAT1 (5861, 1:250 for IF), NPRL2 (37344), phospho-S6K (9234), S6K (2708), phospho-S6 S235/236 (4858, 1:3000 for WB), S6 (2217, 1:5000 for WB), TAX1BP1 (5105), TFEB (4240), TFEB (37785, 1:200 for IF), and phospho-ULK S757 (14202) were from Cell Signaling Technology. Mouse monoclonal anti–S. Typhimurium lipopolysaccharide (clone 1E6, ab8274) and FNIP1 (ab134969) were from Abcam. TFE3 (HPA023881) was from MilliporeSigma. p62 (GP62-C) was from Progen. Galectin-3 (sc-23938) was from Santa Cruz Biotechnology. TFEB (A303-673A, 1:200 for IF in murine cells) was from Bethyl Laboratories. All antibodies were used at a 1:1000 dilution for Western blotting unless otherwise noted.

Goodwin J.M., Walkup WG I.V., Hooper K., Li T., Kishi-Itakura C., Ng A., Lehmberg T., Jha A., Kommineni S., Fletcher K., Garcia-Fortanet J., Fan Y., Tang Q., Wei M., Agrawal A., Budhe S.R., Rouduri S.R., Baird D., Saunders J., Kiselar J., Chance M.R., Ballabio A., Appleton B.A., Brumell J.H., Florey O, & Murphy L.O. (2021). GABARAP sequesters the FLCN-FNIP tumor suppressor complex to couple autophagy with lysosomal biogenesis. Science Advances, 7(40), eabj2485.

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Quantifying Autophagy Markers by RT-qPCR and Western Blot

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SYBR Green real-time quantitative polymerase chain reaction (RT-qPCR) and Western blot were performed as described previously (40 (link)). Primer sequences are available upon request. The following antibodies were used for Western blots: Vps34 (4263; Cell Signaling), p62 (GP62-C; Progen), LC3B (2775; Cell Signaling), phospho-STAT1 (Tyr⁷⁰¹) (7649; Cell Signaling), STAT1 (9172; Cell Signaling), IRF1 (8478; Cell Signaling), IRF7 (696402; Cell Signaling), and actin (A5441; Sigma-Aldrich).

Noman M.Z., Parpal S., Van Moer K., Xiao M., Yu Y., Viklund J., De Milito A., Hasmim M., Andersson M., Amaravadi R.K., Martinsson J., Berchem G, & Janji B. (2020). Inhibition of Vps34 reprograms cold into hot inflamed tumors and improves anti–PD-1/PD-L1 immunotherapy. Science Advances, 6(18), eaax7881.

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