Paraffin-embedded kidney sections were also used for IF studies. The sections were deparaffinized and sequentially incubated with 0.1 M sodium citrate for antigen retrieval, and 3% H2O2 to block endogenous peroxidase activity. Then 2% normal goat serum blocking buffer was used to reduce nonspecific binding. The sections were then incubated with anti-CHOP (Proteintech, 1:200) and anti-GRP78 antibodies (1:500) at 4 °C. After being incubated overnight incubation at 4 °C, the samples were incubated with secondary antibodies for 1 h at 37 °C in the dark. The sections were counterstained using the antifade mounting medium with DAPI (P0131, Beyotime, Shanghai, China) and then observed and photographed under a fluorescence microscope (BX51, Olympus, Tokyo, Japan).
Anti chop
Manufactured by Proteintech
Sourced in United States
Anti-CHOP is a primary antibody that specifically binds to the CHOP protein. CHOP is a transcription factor that plays a role in the cellular stress response. This antibody can be used to detect and quantify CHOP expression in various experimental systems.
Automatically generated - may contain errors
Lab products found in correlation
Anti perk, by Proteintech (5 mentions)
Anti bip, by Proteintech (4 mentions)
Anti p eif2α ser51, by Cell Signaling Technology (3 mentions)
Anti ubiquitin, by Proteintech (3 mentions)
Anti cse, by Proteintech (3 mentions)
Anti eif2α, by Proteintech (3 mentions)
Anti p perk thr980, by Cell Signaling Technology (3 mentions)
P0131, by Beyotime (2 mentions)
Anti dgat1, by Proteintech (2 mentions)
Anti β tubulin, by Proteintech (2 mentions)
Anti caspase 3, by Proteintech (2 mentions)
Pierce bca protein assay kit, by Thermo Fisher Scientific (2 mentions)
Ecl kit, by Thermo Fisher Scientific (2 mentions)
Ab214821, by Abcam (1 mentions)
Ab109199, by Abcam (1 mentions)
Imager 600, by Cytiva (1 mentions)
Anti runx2, by Cell Signaling Technology (1 mentions)
Anti β actin, by Cell Signaling Technology (1 mentions)
Anti p53, by Wuhan Servicebio Technology (1 mentions)
Ecl reagent, by Beyotime (1 mentions)
13 protocols using anti chop
1
Immunodetection of ER Stress Markers
2
Western Blot Analysis of Vascular Smooth Muscle Cells
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Vascular smooth muscle cells and arterial tissues were homogenized with RIPA buffer containing proteinase and phosphatase inhibitors. The protein concentrations were quantified with Pierce™ BCA Protein Assay Kit (Thermo Fisher Scientific) according to the manufacturer's guidelines. Proteins separated by SDS‐PAGE were transferred to PVDF membranes, followed by being incubated with 5% milk overnight at 4°C. Membranes were immunoblotted using specific primary antibodies: anti‐BMP2 (1:1000, ab214821; Abcam), anti‐Runx2 (1:1000, 12556S; Cell Signaling Technology), anti‐SIRT1 (1:1000, 9475S; Cell Signaling Technology), anti‐CHOP (1:1000, 15204–1‐AP; Proteintech), anti‐ATF4 (1:1000, 10835–1‐AP; Proteintech), anti‐p21 (1:1000, ab109199; Abcam), and anti‐β‐actin (1:1000, 4970S; Cell Signaling Technology). Membranes were rinsed with PBST for three times, and then incubated with HRP‐conjugated secondary antibodies. The blots were visualized by the imaging system (Amersham Imager 600) and quantified using Image J software.
3
Western Blot Analysis of Protein Expression in NP Tissue
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Total proteins from human NP tissues or NP cells were lysed using RIPA lysis buffer (Beyotime, China) according to standard methods. Then, proteins were electrophoresed in 10% or 12% SDS‒PAGE gels and transferred to PVDF membranes. Membranes were blocked with 5% nonfat dried milk for 2 h and incubated with anti-GRP78 (Zenbio, China; 1:1000 dilution), anti-CHOP (Proteintech, China, 1:1000 dilution), anti-P53 (Servicebio, China; 1:1000 dilution), anti-P21 (Zenbio, China; 1:1000 dilution), anti-P16 (Cohesionbio, UK; 1:1000 dilution), anti-RB (CST, USA; 1:1000 dilution) and anti-SLC43A3 (Santa, USA, 1:1000 dilution) antibodies at 4 °C overnight. After washing with TBST, the membranes were incubated with horseradish peroxidase (HRP)-conjugated secondary antibodies and visualized using an ECL reagent (Beyotime Biotech, China). GAPDH expression was used for normalization.
4
Protein Expression Analysis Protocol
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All protein samples were quantified by using the BCA reagent (Beyotime). Anti‐CSE, anti‐DGAT1, anti‐SREBP1, anti‐AGPAT3, anti‐CHOP, anti‐eIF2α, anti‐PERK, anti‐BIP, anti‐β‐Tubulin, anti‐SYVN1 and anti‐Ubiquitin were from Proteintech. Anti‐P‐eIF2α (Ser51) and anti‐P‐PERK (Thr980) were from Cell Signalling Technology. Specific bands were recorded via a chemiluminescence detection system (Thermo). The band intensity was conducted by ImageJ tool.
5
Western Blot Analysis of ER Stress Proteins
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Cells were lysed with ice-cold cell lysis buffer. Protein concentrations were determined using a BCA protein assay kit (Pierce, number 23225). Equal aliquots (30 μg) of protein samples were applied to 10% SDS-PAGE gels, transferred to polyvinylidene fluoride (PVDF) membranes, and blocked with 5% skim milk TBST (Tris-buffered Saline Tween-20) buffer. The membranes were incubated with primary antibodies including anti-BIP, anti-PDI, anti-CHOP (1:1000; Proteintech, USA); anti-GAPDH (1:1000; Santa Cruz Biotechnology) at 4°C overnight. Then the membranes were incubated with anti-rabbit or anti-mouse antibodies at room temperature for 1 h. The protein bands were captured and documented through gel image analysis system (ChemiDox XRS, Bio-Rad, USA). The intensities of the protein bands were analyzed by Molecular Imaging Software Version 4.0, which was provided by Kodak 2000 MM System. GADPH protein was used as the internal control.
6
Immunofluorescent Labeling of CHOP, GFAP, and Iba1
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After the section preparation and antigen retrieval, the sections were washed with PBS
and blocked with normal goat serum (SL038, Solarbio Science & Technology) for 15 min
at room temperature. Then, the sections were incubated overnight at 4°C with the primary
rabbit anti-CHOP (1:200, Proteintech) antibody, mouse anti-GFAP (1:50, Santa Cruz
Biotechnology, Santa Cruz, CA, USA) antibody and mouse anti-Iba1 (1:200, Genetex)
antibody. After washing, the sections were incubated in Cy3-conjugated and FITC-conjugated
secondary antibodies (A0516, A5608, 1:200, Beyotime Biotechnology, Haimen, China) for 1 h
at room temperature in the dark. The sections were further treated with DAPI for another
20 min at room temperature. After the washing and sealing, the images were observed using
a fluorescence microscope (DP73, Olympus, Tokyo, Japan).
and blocked with normal goat serum (SL038, Solarbio Science & Technology) for 15 min
at room temperature. Then, the sections were incubated overnight at 4°C with the primary
rabbit anti-CHOP (1:200, Proteintech) antibody, mouse anti-GFAP (1:50, Santa Cruz
Biotechnology, Santa Cruz, CA, USA) antibody and mouse anti-Iba1 (1:200, Genetex)
antibody. After washing, the sections were incubated in Cy3-conjugated and FITC-conjugated
secondary antibodies (A0516, A5608, 1:200, Beyotime Biotechnology, Haimen, China) for 1 h
at room temperature in the dark. The sections were further treated with DAPI for another
20 min at room temperature. After the washing and sealing, the images were observed using
a fluorescence microscope (DP73, Olympus, Tokyo, Japan).
7
Protein Extraction and Western Blot Analysis
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Tissues were homogenized in ice-cold lysis buffer (50 mM Tris HCl, pH 7.5, 0.5% Nonidet P-40, 150 mM NaCl, 2 mM EGTA, 1 mM Na3VO4, 100 mM NaF, 10 mM Na4P2O7, 1 mM phenylmethylsulfonyl fluoride, 10 μg/ml aprotinin, 10 μg/ml leupeptin)11 (link). Tissue extracts were then immunoblotted with the following primary antibodies: anti-p-eIF2α, anti-EIF2α, anti-GCN2, anti-p-HSL, anti-HSL, anti-IRE1α, and anti-p-PKA substrates (1:1000, Cell Signaling Technology, MA, USA); anti-UCP1 and anti-ATF6 (1:1000, Abcam, Cambridge, UK); anti-TH (1:1000, Merck Millipore, Frankfurter, GER); anti-ATF4 and anti-TRB3 (1:500, Santa Cruz Biotechnology, CA, USA), anti-p-GCN2 (1:500, Biorbyt, Cambridge, UK); anti-CHOP, anti-ATF4, anti-PERK, and anti-XBP1s (1:1000, Proteintech, Hubei, P.R.C.); anti-p-IRE1α (1:1000, Epitomics, CA, USA); anti-p-PERK (1:1000, Signalway Antibody, MD, USA); anti-BIP and anti-β-actin (1:2000, Sigma, MO, USA).
8
Antibody-based Protein Expression Analysis
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Anti-HA antibody was from Huaxingbio (Beijing, China); anti-caspase3 antibody was from Cell Signaling Technology (Danvers, MA, USA); anti-caspase 8, anti-caspase 4, anti-cleaved-caspase 3, anti-Bcl-XL, anti-cleaved caspase 8, anti-cytochrome C, anti-CHOP, anti-PARP1, and anti-GRP78 antibodies were all purchased from Proteintech Group, Inc. (Rosemont, IL, USA); anti-caspase 10 antibody was from Bioworld Technology, Inc. (St. Louis Park, MN, USA); anti-β-actin antibody was from Transgen Biotechnology (Beijing, China); anti-GAPDH antibody was from Affinity Biosciences (Cincinnati, OH, USA). Human recombinant IFNalpha-1a was purchased from ProSpec-Tany TechnoGene (Ness Ziona, Israel). Cell counting kit-8 was from Dojindo (Kumamoto, Japan).
9
Analyzing Lung Tumor Protein Expression
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Lung tumor tissues and cells were lysed in RIPA lysis buffer and BCA kit was adopted to test the concentration of total protein (E-BC-K318-M, Elabscience, Wuhan, China). Then, protein was separated, and transferred into PVDF membrane. After blocking with skimmed milk, the membranes were incubated overnight anti-IRE1α (27,528-1-AP, 1:2000), anti-XBP1s (24,868-1-AP, 1:6000), anti-GRP78/BIP (GRP78/BIP, 1:10,000), anti-CHOP (15,204-1-AP, 1:3000), anti-PERK (24,390-1-AP, 1:2000), and anti-β-actin (81,115-1-RR, 1:20,000) (Proteintech, Wuhan, China); anti-E-cadherin (#3195, 1:1000), and anti-N-cadherin (#13,116, 1:1000) (CST, Boston, MA, USA); and Vimentin (sc-373717, 1:5000, Santa Cruz Biotechnology, Inc, Santa Cruz, CA, USA). Second antibody (anti-Rabbit IgG-HRP, SA00001-2, 1:10,000, Proteintech, Wuhan, China) was subsequently incubated for 2 h. Finally, immunoblots were performed by the enhanced chemiluminescence (ECL) kit (34,577, Thermo Fisher Scientific, Waltham, MA, USA), and visualized by Tanon 5200 (Tanon, Shanghai, China).
10
Protein Expression Analysis in Oocyte Development
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Total protein was extracted from GCs, oocytes, and ovaries using RIPA buffer (Solarbio, R0020, China). Protein concentration was assessed by BCA assay (Cellchip, China). Protein extracts separated by SDS-PAGE were blotted with the following primary antibodies: anti-IRE1α (1:1000, CST, 3294), anti-phospho-IRE1 (Ser724, 1:1000, Abcam, ab48187), anti-Bcl2 (1:500, Proteintech, 12789-1-AP), anti-Bax (1:500, Ruiying, RLT0456), anti-JNK (1:500, Proteintech, 51151-1-AP), anti-phospho-JNK (Thr183/Tyr185, 1:1000, CST, 4688), anti-caspase 9 (1:1000, CST, 9508), anti-cleaved caspase 9 (1:1000, CST, 9509) anti-caspase 3, (1:700, Proteintech, 19677-1-AP), anti-cleaved caspase 3 (1:1000, CST, 9661), anti-XBP1 (1:400, Santa Cruz, sc8015), anti-ATF6 (1:400, Santa Cruz, sc166659), anti-BiP/GRP78 (1:1000, Santa Cruz, sc376768), anti-phospho-PERK (Thr980, 1:1000, CST, 3179), anti-PERK (1:1000, CST, 3192), anti-phospho-eIF2α (Ser51, 1:1000, CST, 3398), anti-eIF2α (1:1000, CST, 9722), anti-ATF4 (1:1000, CST, 11815), anti-CHOP (1:500, Proteintech, 15204-1-AP), and TMCO1 antibody (1:1000) [13 (link)].
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