Na9340

Manufactured by GE Healthcare

Sourced in Japan, United Kingdom, United States

The NA9340 is a high-performance liquid chromatography (HPLC) system designed for laboratory analysis. It is capable of separating and detecting a wide range of chemical compounds with precision and accuracy. The core function of the NA9340 is to provide reliable and efficient HPLC analysis for various applications.

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Lab products found in correlation

Na9310, by GE Healthcare (4 mentions) Halt phosphatase inhibitor cocktail, by Thermo Fisher Scientific (3 mentions) Can get signal, by Toyobo (2 mentions) Anti β actin, by Cell Signaling Technology (2 mentions) Trans blot turbo transfer system, by Bio-Rad (2 mentions) Western lightning plus ecl, by PerkinElmer (2 mentions) Ab32072, by Abcam (2 mentions) Ecl kit, by Cytiva (1 mentions) A9522, by Merck Group (1 mentions) Ab8580, by Abcam (1 mentions) Ab1791, by Abcam (1 mentions) Western lightning plus ecl enhanced chemiluminescence reagent, by PerkinElmer (1 mentions) Chemidoc touch imaging system, by Bio-Rad (1 mentions) Pierce ecl plus western blotting substrate, by Thermo Fisher Scientific (1 mentions) A2066, by Merck Group (1 mentions) Ab207298, by Abcam (1 mentions) Catalase activity assay kit, by Abcam (1 mentions) 8 ohdg elisa kit, by Abcam (1 mentions) Anti gapdh, by Cell Signaling Technology (1 mentions) Mitosox red mitochondrial superoxide indicator, by Thermo Fisher Scientific (1 mentions)

14 protocols using na9340

Antithrombin Western Blot Analysis

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Plasma antithrombin was evaluated by Western blot using different electrophoretic conditions described elsewhere.11 The blotting was performed by using rabbit anti‐human antithrombin polyclonal antibody (A9522, Sigma‐Aldrich) followed by donkey anti‐rabbit IgG‐horseradish peroxidase conjugate (NA9340, GE Healthcare, Barcelona, Spain) as secondary antibody.
Detection was carried out with an ECL kit (Amersham Biosciences, Piscataway, NJ, USA) in the ImageQuant LAS4000mini equipment (Exon Biotec, GE Healthcare).

de la Morena‐Barrio M.E., López‐Gálvez R., Martínez‐Martínez I., Asenjo S., Sevivas T.S., López M.F., Wypasek E., Entrena L., Vicente V, & Corral J. (2017). Defects of splicing in antithrombin deficiency. Research and Practice in Thrombosis and Haemostasis, 1(2), 216-222.

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Chromatin Remodeling in Stressed Worms

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Stressed P0 parents (at day 2 adulthood) and the unstressed F1 descendants (at day 1 adulthood) were washed by M9 buffer, and frozen in liquid nitrogen. Worm extracts were lysed in sample buffer (2% SDS, 10% glycerol, 5% β-mercaptoethanol, 62.5 mM Tris-HCl pH 6.8, 0.001% bromophenol blue), followed by sonication. After centrifugation, supernatants were collected and boiled for 10 min. Samples were resolved by SDS–polyacrylamide gel electrophoresis (14% polyacrylamide gels) and subjected to immunoblotting according to a standard protocol using primary antibodies (anti-Histone H3 (Abcam ab1791, 1:1,000), anti-Histone H3K4me3 (Abcam ab8580, 1:1,000) and an anti-rabbit secondary antibody (GE Healthcare NA9340, 1:10,000)). Signals were detected with Western Lightning Plus-ECL Enhanced Chemiluminescence reagent (PerkinElmer). Band intensities were measured using ImageJ software. All uncropped western blots can be found in Supplementary Fig. 8.

Kishimoto S., Uno M., Okabe E., Nono M, & Nishida E. (2017). Environmental stresses induce transgenerationally inheritable survival advantages via germline-to-soma communication in Caenorhabditis elegans. Nature Communications, 8, 14031.

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Western Blot Analysis of FoxM1 in Islet Samples

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Islet samples were boiled in Laemmli buffer containing 10 mM dithiothreitol at 100°C for 5 min, subjected to sodium dodecyl sulfate‐polyacrylamide gel electrophoresis and transferred onto nitrocellulose membranes, then blocked in Tris‐buffered saline with 3% fetal bovine serum. Nitrocellulose membranes were incubated with primary antibodies to FoxM1 (ab207298, Abcam) at a 1:2,000 dilution and Actin (A2066, Sigma) at a 1:5,000 dilution, and then incubated with a secondary horseradish peroxidase‐conjugated antibody (NA9340; GE Healthcare, Tokyo, Japan) at a 1:10,000 dilution. These antibodies were dissolved in Can Get Signal (Toyobo). As blottings of FoxM1 and actin were performed using the same membrane. Quantitative data were obtained by employing the Pierce ECL Plus Western Blotting Substrate (Thermo Fisher Scientific, Waltham, MA, USA) and the ChemiDoc Touch Imaging System (Bio‐rad Laboratories). Relative intensities were standardized employing actin intensity as the invariant control using ImageJ Fiji. Uncropped images are presented in Figure S1.

Kohata M., Imai J., Izumi T., Yamamoto J., Kawana Y., Endo A., Sugawara H., Seike J., Kubo H., Komamura H., Sato T., Hosaka S., Munakata Y., Asai Y., Kodama S., Takahashi K., Kaneko K, & Katagiri H. (2022). Roles of FoxM1‐driven basal β‐cell proliferation in maintenance of β‐cell mass and glucose tolerance during adulthood. Journal of Diabetes Investigation, 13(10), 1666-1676.

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Investigating H. pylori-Induced Cellular Stress

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Primary antibodies we used were anti-H. pylori (ab7788, Abcam), anti-HSPA8 (10654-1-AP, Proteintech), anti-HSPA1A (10995-1-AP, Proteintech), anti-HSPA1B (25405-1-AP, Proteintech), anti-BRCA1 (22362-1-AP, Proteintech), anti-SUZ12 (20366-1-AP, Proteintech), anti-FANCM (12954-1-AP, Proteintech), anti-ZO-1 (21773-1-AP, Proteintech), anti-Claudin 1 (13050-1-AP, Proteintech), anti-Occludin (66378-1-Ig, Proteintech), anti-β-Actin (4967S, Cell Signaling Technology), and anti-GAPDH (2118S, Cell Signaling Technology). Secondary antibodies used included anti-mouse conjugated to horseradish peroxidase (A2304, Sigma-Aldrich) and anti-rabbit conjugated to horseradish peroxidase (NA9340, GE Healthcare) for western blotting; anti-rabbit conjugated to Alexa Fluor 568 (A11011) and anti-mouse conjugated to Alexa Fluor 488 (A21202) were from Life Technology for confocal microscopy.
The Catalase Activity Assay Kit was purchased from Abcam (ab83464). DSS (160110) was from MP Biomedicals (Shanghai) Co. Ltd. MitoSox Red mitochondrial superoxide indicator was from Thermo Fisher Scientific (M3608). The 8-OHdG ELISA Kit was from Abcam (ab201734).

Hu W., Zhai Z.Y., Huang Z.Y., Chen Z.M., Zhou P., Li X.X., Yang G.H., Bao C.J., You L.J., Cui X.B., Xia G.L., Ou Yang M.P., Zhang L., Wu W.K., Li L.F., Zhang Y.X., Xiao Z.G, & Gong W. (2024). Dual RNA sequencing of Helicobacter pylori and host cell transcriptomes reveals ontologically distinct host-pathogen interaction. mSystems, 9(4), e00206-24.

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Analyzing Cohesin Complex Protein Levels

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Protein was extracted from equal cell numbers in a high urea buffer (48% urea, 15 mM Tris-HCl, pH 7.5, 8.7% glycerin, 1% SDS, 0.004% bromophenol blue, and 143 mM β-mercaptoethanol). Protein levels were measured using the Bradford assay. Equal amounts of protein were loaded on NuPAGE Novex 4–12% Bis-Tris protein gel (NP0321; Life Technologies) and run with NuPAGE MOPS buffer (NP0001; Life Technologies). Proteins were transferred to Immobilon P (IPVH00010; EMD Millipore). Membrane was blocked in 5% nonfat milk in TBST and incubated with antibodies for Rad21 (H-210; Santa Cruz Biotechnology, Inc.), Smc1a (A300-055A; Bethyl Laboratories, Inc.), Smc3 (ab9263; Abcam), Stag2 (J-12; Santa Cruz Biotechnology, Inc.) and Actin B (C4; EMD Millipore). Secondary antibodies used were HRP conjugated (NA9340 and NA931; GE Healthcare). Blot was visualized using ECL (PI34077 and PI34095; Thermo Fisher Scientific).

Mullenders J., Aranda-Orgilles B., Lhoumaud P., Keller M., Pae J., Wang K., Kayembe C., Rocha P.P., Raviram R., Gong Y., Premsrirut P.K., Tsirigos A., Bonneau R., Skok J.A., Cimmino L., Hoehn D, & Aifantis I. (2015). Cohesin loss alters adult hematopoietic stem cell homeostasis, leading to myeloproliferative neoplasms. The Journal of Experimental Medicine, 212(11), 1833-1850.

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Exosomal Protein Extraction and Analysis

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Total proteins were extracted from purified exosomes using RIPA buffer, sonication and vortexing. The exosomal proteins (10 μg) were run on a Mini-Protean TGX precast gel (Any kD, Bio-Rad Laboratories, Hercules, CA, USA) and blotted to Trans-Blot Mini PVDF membranes using the Trans-Blot Turbo™ Transfer system (Bio-Rad). Blots were incubated with blocking buffer (5% non-fat milk/PBST) overnight at 4°C. OVA was detected using an anti-OVA antibody (1:1,000; 0220-1682, AbD Serotec, Kidlington, UK), together with a donkey-anti-rabbit secondary antibody (1:10,000; NA9340, GE Healthcare, Little Chalfont, UK). Alix was detected using an anti-Alix antibody (1:1000; 2171 (3A9), Cell Signaling technology, Danvers MA, US), together with a sheep-anti-mouse secondary antibody (1:2000; NA9310V, GE Healthcare, Little Chalfont, UK) and visualized by enhanced chemiluminescence (GE Healthcare), the ChemiDoc™ MP Imaging System and Image Lab™ software version 4.1 (both from Bio-Rad).

Hiltbrunner S., Larssen P., Eldh M., Martinez-Bravo M.J., Wagner A.K., Karlsson M.C, & Gabrielsson S. (2016). Exosomal cancer immunotherapy is independent of MHC molecules on exosomes. Oncotarget, 7(25), 38707-38717.

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Western Blot Analysis of ALDH1/2, cMYC, and Cleaved Caspase-3

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Western blot analyses were carried out as described previously (Zhu et al. 2019 (link)). After washing with phosphate buffered saline (PBS), cultured cells treated with vehicle or DAC were homogenized in a solution with 50 mM tris buffer, 150 mM NaCl, 1 mM ethylenediaminetetraacetic acid, and 1% NP40 and proteinase (Roche Diagnostics, GmbH, Mannheim, Germany)/phosphatase inhibitors (Halt Phosphatase Inhibitor Cocktail, Thermo Scientific). After removal of cell debris, cell lysates were used for Western blot analysis. The antibodies against ALDH1/2 (sc-166362) were purchased from Santa Cruz Biotechnology, Inc. The antibody against cMYC (ab32072) was from Abcam. Antibody against cleaved caspase-3 (#9664) was from Cell Signaling. After being washed, the membranes were incubated with horseradish peroxidase-conjugated anti-mouse immunoglobulin G (NA310 GE Healthcare) or anti-rabbit IgG (NA9340 GE Healthcare) as the secondary antibody and subsequently detected by means of an ECL system (PerkinElmer, Waltham, MA, USA). Band intensities were quantified by the ImageJ software (ImageJ 1.53a; Wayne Rasband, NIH).

Runyon G.T., Bear D.G, & Lohman T.M. (1990). Escherichia coli helicase II (UvrD) protein initiates DNA unwinding at nicks and blunt ends.. Proceedings of the National Academy of Sciences of the United States of America, 87(16), 6383-6387.

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Western Blot Analysis of Proteins

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Total protein lysates were extracted from whole-cell lines in phosphate-buffered saline. The Pierce BCA Protein Assay Kit (Cat#23225, Thermo Fisher) was used to determine concentration. All proteins were separated by sodium dodecyl sulfate–polyacrylamide gel electrophoresis, and the proteins in the gel were then transferred to polyvinylidene difluoride blotting membrane (A29532146, P 0.45 GE Healthcare Life Science, Chicago, IL, USA). Trans-Blot Turbo Cassette (Bio-Rad, Hercules, CA, USA) was used for blotting. For blocking, 5% skimmed milk was used. Primary antibodies including anti-YTHDF1 (1:1000 dilution, 17479-1-AP, Proteintech), YTHDF2 (1:1000 dilution, ab170118, Abcam), PD-L1 (1:1000 dilution, A1935; ABclonal, Tokyo, Japan), and GAPDH (1:1000 dilution, Ab8245; Abcam) were incubated overnight at 4°C. Secondary antibodies for rabbit (1:20,000 dilution, NA9340; GE Healthcare Life Science) or mouse (1:20,000 dilution, NA9310; GE Healthcare Life Science) were incubated at room temperature for 1 hour. To visualize the band, we used enhanced chemiluminescence (Pierce ECL Plus Substrate, Thermo Scientific). The photo was detected by a chemi-doc (Bio-Rad). Signal intensity was quantified using Image Labo Software (Bio-Rad).

Tsuchiya K., Yoshimura K., Inoue Y., Iwashita Y., Yamada H., Kawase A., Watanabe T., Tanahashi M., Ogawa H., Funai K., Shinmura K., Suda T, & Sugimura H. (2021). YTHDF1 and YTHDF2 are associated with better patient survival and an inflamed tumor-immune microenvironment in non–small-cell lung cancer. Oncoimmunology, 10(1), 1962656.

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Western Blot Analysis of Tumor Proteins

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Western blot analyses were carried out as described previously (Zhu et al. 2014 (link)). Tumor tissues or cultured cells were washed with phosphate-buffered saline and homogenized in a solution with 50 mM Tris buffer, 150 mM NaCl, 1 mm ethylenediaminetetraacetic acid, 1% NP40 and proteinase (Roche Diagnostics, GmbH, Mannheim, Germany)/phosphatase inhibitors (Halt Phosphatase inhibitor cocktail, Thermo Scientific). After centrifuge at 14000 rpm for 5 minutes, lysates were used for Western blot analysis. The antibodies against MYC were used (ab32072) from Abcam, phosphorylated RB (S780) (#9307), RB (#9309), Bim (#2933), p-Erk½ (#9101), total-ERK (#9102), p27 (#3686), PARP (#9542) and GAPDH (#2118) from Cell Signaling. After being washed, the membranes were incubated with horseradish peroxidase-conjugated anti-mouse immunoglobulin G (Gillings, et al.) (NA9310 GE Healthcare) or anti-rabbit IgG (NA9340 GE Healthcare) as the secondary antibody and subsequently detected by means of an ECL system (Western Lightning® Plus-ECL, PerkinElmer, Waltham, MA). Band intensities were quantified by the ImageJ software (ImageJ 1.48v; Wayne Rasband, NIH).

Zhu X., Park S., Lee W.K, & Cheng S.Y. (2019). Potentiated Anti-tumor Effects of BETi by MEKi in Anaplastic Thyroid Cancer. Endocrine-related cancer, 26(9), 739-750.

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Exosome Protein Extraction and Analysis

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Purified exosomes were treated with lysis buffer (20 mM Tris HCL/1% SDS) and subjected to sonication and vortexing, with protein concentration determined using a BCA protein assay kit (Thermo Scientific Pierce, Rockford, IL, USA) according to manufacturer’s recommendations. The exosomal proteins (50 μg) were run on a Mini-Protean TGX precast gel (Any kD, Bio-Rad Laboratories, Hercules, CA, USA) and blotted to Trans-Blot Mini PVDF membranes using the Trans-Blot® Turbo™ Transfer system (Bio-Rad). Blots were processed using the Snap i.d. protein detection system (Millipore, Billerica, MA, USA) according to manufacturer’s recommendations. Melan-A was detected using an anti-Melan-A antibody (1:250; FL-118, sc-28871, Santa Cruz Biotechnology, Santa Cruz, CA, USA) together with a donkey-anti-rabbit secondary antibody (1:10000; NA9340, GE Healthcare, Little Chalfont, UK) and visualised by enhanced chemiluminescence (GE Healthcare), the VersaDoc™ 4000 MP Imaging System and Quantity One® software (both from Bio-Rad).

Eldh M., Olofsson Bagge R., Lässer C., Svanvik J., Sjöstrand M., Mattsson J., Lindnér P., Choi D.S., Gho Y.S, & Lötvall J. (2014). MicroRNA in exosomes isolated directly from the liver circulation in patients with metastatic uveal melanoma. BMC Cancer, 14, 962.

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