Western blotting has previously been described (18 (link)). Anti-pEIF2αS51, anti-EIF2α, anti-ATF4, anti-GRP78, anti-IRE1α, anti-acetylated-α-tubulin, anti-BCL2, anti-BCLXL, anti-PUMA, anti-BID, anti-BIM (Cell Signaling Technology, Beverly, MA, USA) and anti-pIRE1αS724 (Abcam, Cambridge) were used in conjunction with a HRP-conjugated anti-rabbit secondary antibody (Amersham, Buckinghamshire, UK). Anti-caspase-8 (12F5; Alexis, San Diego, CA, USA), anti-CHOP (Cell Signaling Technology), anti-ATF6 (Abcam), anti-MCL1 (BD pharmingen, Oxford, UK) and anti-NOXA (Abcam) mouse monoclonal antibodies were used in conjunction with a horseradish peroxidase–conjugated anti-mouse secondary antibody (Amersham).
Anti noxa
Manufactured by Abcam
Sourced in United Kingdom, United States
Anti-NOXA is a laboratory reagent used for research purposes. It is an antibody that specifically binds to the NOXA protein, which is a pro-apoptotic member of the Bcl-2 family. This antibody can be used in various techniques, such as Western blotting, immunoprecipitation, and immunohistochemistry, to detect and study the expression and localization of NOXA in biological samples.
Automatically generated - may contain errors
Lab products found in correlation
Anti bim, by Abcam (2 mentions)
Anti chop, by Cell Signaling Technology (1 mentions)
Anti ire1α, by Cell Signaling Technology (1 mentions)
Anti atf6, by Abcam (1 mentions)
Anti mcl 1, by BD (1 mentions)
Anti bcl 2, by Cell Signaling Technology (1 mentions)
Anti atf4, by Cell Signaling Technology (1 mentions)
Anti grp78, by Cell Signaling Technology (1 mentions)
Anti puma, by Cell Signaling Technology (1 mentions)
Anti peif2αs51, by Cell Signaling Technology (1 mentions)
Anti acetylated α tubulin, by Cell Signaling Technology (1 mentions)
Hrp conjugated anti rabbit secondary antibody, by Cytiva (1 mentions)
Horseradish peroxidase conjugated anti mouse secondary antibody, by Cytiva (1 mentions)
Anti bim, by Cell Signaling Technology (1 mentions)
Anti bcl xl, by Cell Signaling Technology (1 mentions)
Anti bid, by Cell Signaling Technology (1 mentions)
Anti eif2α, by Cell Signaling Technology (1 mentions)
Anti actin, by Merck Group (1 mentions)
Anti active caspase 3, by Abcam (1 mentions)
Anti hdac6, by Santa Cruz Biotechnology (1 mentions)
7 protocols using anti noxa
1
Comprehensive Protein Expression Analysis
2
Protein Expression Analysis in Cell Lysates
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After treatment under the indicated conditions, cells were washed with PBS, suspended in radioimmunoprecipitation (RIPA) buffer, incubated on ice for 15 min, and centrifuged at 20380 g for 12 min. Whole cell lysate was subjected to Western blot analysis as described previously.7 To analyze the expression of ubiquitinated proteins in the detergent‐insoluble fractions (pellets obtained after the protein extraction using RIPA buffer), the pellets were washed with PBS, lysed using Extraction buffer 4 in the WSE‐7421 EzSubcell Extract kit (ATTO, Tokyo, Japan), and then subjected to Western blot analysis. The following antibodies were used: anti‐cyclin D1, anti‐cyclin‐dependent kinase (CDK) 4, anti‐glucose‐regulated protein (GRP) 78, anti‐ubiquitin, anti‐histone deacetylase (HDAC) 1, anti‐HDAC3, and anti‐HDAC6 from Santa Cruz Biotechnology (Santa Cruz, CA, USA); anti‐cleaved poly(ADP‐ribose) polymerase (PARP), anti‐HSP70, anti‐endoplasmic reticulum resident protein (ERP) 44, and anti‐endoplasmic oxidoreductin‐1‐like protein (Ero1‐L) from Cell Signaling Technology (Danvers, MA, USA); anti‐active caspase 3, anti‐NOXA, and anti‐acetylated histone from Abcam (Cambridge, UK); and anti‐actin from Millipore (Billerica, MA, USA).
3
Apoptosis Regulator Expression Analysis
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Cell lysates were prepared in RIPA buffer (Sigma) plus protease inhibitor cocktail (Roche). 20 μg of total protein was resolved on a 12 % SDS polyacrylamide gel and probed with anti-BCL-XL (Epitomics, Burlingame, CA), anti-MCL-1 (Epitomics), anti-BCL-2 (BD), anti-BIM (Epitomics), anti-actin and anti-NOXA (Abcam, Cambridge, MA). Antibody against tubulin (Santa Cruz Biotechnology, Inc., Santa Cruz, CA) was used as a loading control.
4
Immunofluorescence Staining of HIF-1α, Noxa, and Bid
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BE(2)-C cells were seeded in 48-well plates and labeled with the following primary antibodies: anti-HIF-1α (Abcam, 1:500), anti-noxa (Abcam, 1:500) and anti-bid (Abcam, 1:500). Primary antibodies were labeled using fluorescein isothiocyanate (FITC)-conjugated anti-rabbit secondary antibodies (Jackson, 1:1000). The cell nuclei were stained using 4′,6-diamidino-2-phenylindole (DAPI) (Beyotime, China).
5
Comprehensive Western Blotting Analysis
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Western blotting analysis was performed following a previously described method 20 (link). Briefly, cells were lysed in radioimmunoprecipitation assay (RIPA) buffer (Sigma-Aldrich, #R0278) containing protease inhibitor (Sigma-Aldrich, P8340). Equal amounts of total proteins were loaded and resolved by SDS-PAGE gels, followed by transferring to a PVDF membrane, blocking with 5% milk, and probing with primary antibodies. The following primary antibodies were used: anti-CtBP1 (BD Bioscience, USA, #612042), anti-CtBP1 (phospho Ser422) (GeneTex, USA, #GTX55356), anti-CtBP2 (BD Bioscience, #612044), anti-HIPK2 (Cell Signaling, USA, #5091S), anti-BIM (Abcam, China, #ab170589), anti-BIK (Abcam, #ab52182), anti-BAX (Abcam, #ab3191), anti-NOXA (Abcam, #114C307), anti-CASP3 (Sigma-Aldrich, #C9598), anti-CASP7 (Sigma-Aldrich, #C1104), anti-CASP9 (Abcam, #ab184786), anti-p300 (Sigma-Aldrich, #P2859), anti-FOXO3a (Sigma-Aldrich, #V38041), and anti-GAPDH (Thermo Fisher Scientific, #MA5-15738-BTIN). The protein signals were visualized using an ECL detection kit (Sigma-Aldrich, #GERPN2109).
6
Western Blot Analysis of Apoptosis Proteins
7
Histological Examination of Lung Tissue
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For histological examination, lung tissues were stained sequentially with haematoxylin and eosin, and Masson's trichrome (Sigma‐Aldrich, St. Louis, MO, USA) staining was carried out as previously described 15 . Immunohistochemical staining was carried out using anti‐Noxa (Abcam, Cambridge, MA, USA) and anti‐8‐OHdG (Genox, Baltimore, MD, USA) antibodies. A terminal deoxynucleotidyl transferase‐mediated deoxyuridine‐5‐triphosphate‐biotin nick end labelling (TUNEL) assay (ApopTag; Millipore, Billerica, MA, USA) was performed according to the manufacturer's instructions.
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