Primary antibodies used for Western blotting and immunostaining were chicken anti-HMGCS2 (Genway, San Diego, CA), rabbit anti-HMGCS2 (AVIVA, San Diego, CA; epitope different from the Genway antibody), chicken anti-MCT1 (Chemicon, Temecula, CA), rabbit anti-MCT1, goat anti-PPARα (Santa Cruz Biotechnology, Dallas, TX), rabbit anti–α-smooth muscle actin, rabbit anti-prohibitin (Abcam, Cambridge, UK), rabbit anti–cytochrome c, rabbit anti-Glut4, rabbit anti–COX IV, rabbit anti-ACC, rabbit anti–phospho-ACC, rabbit anti-AMPKα, rabbit anti–P-AMPKα, rabbit anti-AMPKβ1, rabbit anti–P-AMPKβ1 (Cell Signaling Technology, Danvers, MA), rat anti-Hsc70 (Enzo Life Sciences, Helsinki, Finland), rat anti-K8 (Troma I Developmental Studies Hybridoma Bank, Iowa City, IA), and mouse anti-tubulin (Sigma-Aldrich, Munich, Germany). Secondary antibodies used for Western blotting were anti-mouse immunoglobulin G (IgG)–horseradish peroxidase (HRP), anti-rat IgG-HRP (GE Healthcare, Buckinghamshire, UK), anti-rabbit IgG-HRP (Cell Signaling Technology), anti-chicken IgG-HRP (Genway), and anti-goat IgG-HRP (Cell Signaling Technology). Secondary antibodies used for immunostaining were anti-rabbit Alexa Fluor 488 and anti-rat Alexa Fluor 546 (Life Technologies, Carlsbad, CA). Nuclei were stained with Toto-3 (Life Technologies) or DRAQ5 (Cell Signaling Technology).
Rat anti hsc70
Manufactured by Enzo Life Sciences
Sourced in United States
The Rat anti-Hsc70 is a primary antibody that recognizes the Hsc70 (heat shock cognate 70 kDa protein) protein. Hsc70 is a constitutively expressed member of the heat shock protein 70 family and functions as a molecular chaperone involved in the folding and trafficking of cellular proteins.
Automatically generated - may contain errors
Lab products found in correlation
Draq5, by Cell Signaling Technology (2 mentions)
Rabbit anti acc, by Cell Signaling Technology (1 mentions)
Toto 3, by Thermo Fisher Scientific (1 mentions)
Rabbit anti ampkα, by Cell Signaling Technology (1 mentions)
Rabbit anti α smooth muscle actin, by Abcam (1 mentions)
Anti rat alexa fluor 546, by Thermo Fisher Scientific (1 mentions)
Rabbit anti mct1, by Santa Cruz Biotechnology (1 mentions)
Rabbit anti p ampkα, by Cell Signaling Technology (1 mentions)
Mouse anti tubulin, by Merck Group (1 mentions)
Alexa fluor 488 anti rabbit, by Thermo Fisher Scientific (1 mentions)
Anti rabbit igg hrp, by Cell Signaling Technology (1 mentions)
Rabbit anti iκbα, by Santa Cruz Biotechnology (1 mentions)
Mouse anti k7 rck 105, by Progen Biotechnik (1 mentions)
Rat anti k8, by Developmental Studies Hybridoma Bank (1 mentions)
Alexa 488 anti rat, by Thermo Fisher Scientific (1 mentions)
Rabbit anti ki67, by Abcam (1 mentions)
Poly l lysine (pll), by Fujifilm (1 mentions)
Alexa fluor 488 or 594, by Thermo Fisher Scientific (1 mentions)
Paraphenylenediamine, by Roche (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Roche (1 mentions)
4 protocols using rat anti hsc70
1
Comprehensive Antibody Characterization for Western Blotting and Immunostaining
2
Immunodetection of Cytoskeletal Proteins
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Primary antibodies used for Western blotting and immunofluorescence staining were mouse anti-K7 (RCK-105; Progen, Heidelberg, Germany), rat anti-K8 and rat anti-K19 (Troma I and Troma III, respectively; Developmental Studies Hybridoma Bank, Iowa, IA, USA), rabbit anti-K8 (273) and rabbit anti-K18 (275; kind gifts from J.E. Eriksson), rabbit anti-K20 (It-Ks 20.10; Epitomics, Burlingame, CA, USA), rat anti-Hsc70 (Enzo Life sciences; Farmingdale, NY, USA), mouse anti-K8 pS74 (LJ4; kind gift from M.B. Omary), rabbit anti-Ki67 (Abcam, Cambridge, MA, USA), rat anti-HSF2 (Abcam) and rabbit anti-IκB-α (Santa Cruz Biotechnology; Dallas, TX, USA). Secondary antibodies used for Western blotting were HRP-conjugated anti-mouse (GE healthcare, Little Chalfont, UK), anti-rat (GE healthcare and Cell Signaling Technology, Danvers, MA, USA) and anti-rabbit (Cell Signaling Technology) IgG antibodies. Secondary antibodies used for immunofluorescence staining were Alexa 488/Alexa 546 anti-mouse, Alexa 488 anti-rat and Alexa 488 anti-rabbit antibodies (Invitrogen, Carlsbad, CA, USA). Nuclei were stained with DRAQ5 (Cell Signaling Technology).
3
Immunocytochemistry Protocol for Heat Shock and Proteasome Inhibition
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Cells were grown on coverslips coated with poly-L-lysine (Wako). After treatment with or without heat shock or MG132 as indicated in each figure caption, cells were washed with PBS (137 mM NaCl, 2.7 mM KCl, 8 mM Na2HPO4, and 1 mM KH2PO4), fixed in 2% formaldehyde/PBS (Polysciences) for 15 min at 37°C and then incubated with 50 mM glycine/HMK (20 mM HEPES [pH 7.5], 1 mM MgCl2, and 100 mM KCl) for 5 min at room temperature. After permeabilization with 0.5% Triton X-100/HMK and blocking with 3% skim milk/PBS, cells were incubated with primary antibodies for 1 h at room temperature and detected with secondary antibodies conjugated with Alexa Fluor 488 or 594 (Thermo Fisher Scientific). Coverslips were mounted in PPDI (80% glycerol in PBS and 1 mg/ml paraphenylenediamine [11873580001; Roche]). DNA was counterstained with DAPI (Roche) or DRAQ5 (DR50050; Biostatus). Images were captured with an Olympus BX51 microscope (Figs 1A and B , 3A , and 5A ) or FV1200 confocal microscope (Fig 1C ).
The following antibodies were used as primary antibodies at the indicated dilutions: rat anti-Hsc70 (1:500; 1B5, Enzo Life Sciences) and mouse anti-Hsc70/Hsp70 (1:3,000; 1H5–1, Kose et al, 2012 (link)).
The following antibodies were used as primary antibodies at the indicated dilutions: rat anti-Hsc70 (1:500; 1B5, Enzo Life Sciences) and mouse anti-Hsc70/Hsp70 (1:3,000; 1H5–1, Kose et al, 2012 (link)).
4
Western Blot Analysis of Protein Expression
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Cells were harvested 48 h after transfection and lysed in RIPA (0.1% SDS, 1% Triton X-100, and 0.5% Na deoxycholate in PBS) with Complete EDTA-free protease inhibitor (Roche). Samples were run on tris-glycine gels and transferred to nitrocellulose membranes, then probed with one of the following antibodies: mouse anti-HA (1:1000, Covance), rat anti-Hsc70 (1:5000, Enzo Life Sciences), or rabbit anti-VR1-N (1:1000, Neuromics), followed by an ECL-optimized HRP-conjugated secondary antibody (GE Life Sciences).
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