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Biomax transcreen le

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The BioMax TranScreen LE is a laboratory equipment product designed for imaging and analysis. It provides efficient and reliable performance in various applications. The core function of the BioMax TranScreen LE is to capture and digitize images of biological samples, enabling researchers to analyze and document their findings.

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Lab products found in correlation

Nec290e050uc, by PerkinElmer (2 mentions) Qubit fluorometer, by Thermo Fisher Scientific (2 mentions) Tri reagent, by Thermo Fisher Scientific (1 mentions) Trizol reagent, by Thermo Fisher Scientific (1 mentions) Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (1 mentions) Thrombin, by Merck Group (1 mentions) 35s methionine cysteine, by MP Biomedicals (1 mentions) 3h palmitic acid, by PerkinElmer (1 mentions) Image lab software, by Bio-Rad (1 mentions) Biomax mr film, by Kodak (1 mentions) Biomax ms film, by Kodak (1 mentions)

6 protocols using biomax transcreen le

1

Methionine Pulse-Chase Assay in HeLa Cells

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HeLa cells were electro-transformed twice with siRNAs as described above and grown in 12-well plates. Cells were incubated in methionine-free DMEM (Invitrogen) for 30 min at 37°C, labeled for 15 min with L-methyl 3H methionine (50 mCi/ml) and rinsed twice with DMEM containing unlabeled methionine. They were then incubated for 0 min (i.e. immediately rinsed with ice-cold phosphate buffered saline (PBS)), 30 min, 1 h, 1 h 30 min, 2 h or 2 h 30 min in DMEM containing methionine. After the corresponding chase time, cells were rinsed twice with ice-cold PBS and immediately lysed with 1 ml Trizol reagent (Invitrogen). Total RNAs were extracted with Tri reagent as described below and their concentration was determined with a Qubit fluorometer (Thermo Fisher Scientific). RNA samples were separated on a 1.2% agarose gel and passively transferred to a nylon membrane as described below. The membrane was exposed for 4–6 days to Biomax KODAK TM MS films through a KODAK TM BioMax TranScreen LE.
Montellese C., Montel-Lehry N., Henras A.K., Kutay U., Gleizes P.E, & O’Donohue M.F. (2017). Poly(A)-specific ribonuclease is a nuclear ribosome biogenesis factor involved in human 18S rRNA maturation. Nucleic Acids Research, 45(11), 6822-6836.
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2

FVIII Activation by Thrombin Analysis

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Twenty hours after transfection, COS-1 cells transfected with PMT2-FVIII plasmids were metabolically labeled with [35S]-methionine/cysteine (250 μCi/mL in methionine/cysteine-free DMEM) (MP Biomedicals) for 45 minutes, followed by a 30-minute incubation in complete medium. Cell extracts were prepared by lysis in the NP-40 lysis buffer and immunoprecipitated with anti-FVIII antibody coupled to CL-4B sepharose. Immunoprecipitated proteins were washed with NP-40 lysis buffer and PBS, and resuspended in 50 mM Tris-Hcl pH 7.5, 150 mM NaCl, 2.5 mM CaCl2 and 5% glycerol (buffer A). Immunoprecipitated FVIII in buffer A were divided into two aliquots for incubation in the absence or presence of 5 U/ml thrombin (Sigma) at 37 °C for 30 minutes. The resulting samples were separated in a 12% SDS-PAGE gel and visualized by exposing to an X-ray film using a Kodak Biomax Transcreen LE.
Wei W., Zheng C., Zhu M., Zhu X., Yang R., Misra S, & Zhang B. (2017). Missense mutations near the N-glycosylation site of the A2 domain lead to various intracellular trafficking defects in coagulation factor VIII. Scientific Reports, 7, 45033.
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3

SidJ Glutamylase Activity Assay

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0.1 μg His6-SdeA, 1 μg GST-SidJ were incubated in a 25 μL reaction system containing 50 mM Tris-HCl (pH 7.5) and 1 mM DTT, 5 mM MgCl2, 1 mM L-glutamate. 1 mM ATP and 1 μM CaM for 2 h at 37°C. To measure the glutamylase activity of SidJ using 14C-glutamate, 2 μg His6-SdeA and 0.5 μg GST-SidJ were incubated in a 25 μL reaction system containing 50 mM Tris-HCl (pH 7.5), 1 mM DTT, 5 mM MgCl2, 1 μCi 14C-L-glutamate (Perkin Elmer cat# NEC290E050UC), 1 mM ATP and 1 μM CaM for 2 h at 37°C. Products were resolved by SDS-PAGE gel and stained with Coomassie brilliant blue. Gels were then dried and signals were detected with x-ray films with a BioMax TranScreen LE (Kodak) for 3 d in −80°C.
Gan N., Zhen X., Liu Y., Xu X., He C., Qiu J., Liu Y., Fujimoto G.M., Nakayasu E.S., Zhou B., Zhao L., Puvar K., Das C., Ouyang S, & Luo Z.Q. (2019). Regulation of phosphoribosyl ubiquitination by a calmodulin-dependent glutamylase. Nature, 572(7769), 387-391.
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4

Quantifying CFTR Palmitoylation in Cells

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Cells were grown in 6-well or 10 cm plates. Prior to study, tissue culture medium was replaced with DMEM supplemented with 2% dialyzed FBS and 1% non-essential amino acids for 1 h at 37°C. Following serum deprivation, cells were labeled with 500 μCi/ml 3H-palmitic acid (PerkinElmer, Waltham, MA) for 4 h in DMEM as above, rinsed, and collected in cold PBS with lysis in RIPA buffer. CFTR was immunoprecipitated using 24–1 or 10B6.2 antibody conjugated to protein A/G agarose (Thermo Scientific Pierce Protein Biology Products, Rockford, IL), followed with rinsing and release by 37°C incubation in 4X Laemmli sample buffer containing 10% β-mercaptoethanol. Protein was separated by SDS-PAGE (6% acrylamide) and CFTR detected using autoradiography (Biomax MR film (Kodak) for 3–6 months though a Kodak Biomax transcreen-LE intensifying screen at −80°C). Quantitation was performed with ImageLab software (Bio-Rad Laboratories, Hercules, CA).
McClure M.L., Wen H., Fortenberry J., Hong J.S, & Sorscher E.J. (2014). S-palmitoylation regulates biogenesis of core glycosylated wild-type and F508del CFTR in a post-ER compartment. The Biochemical journal, 459(2), 417-425.
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5

In vitro Hypusination Assay

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In vitro hypusination assay was performed using our previously published procedure [17 (link)]. Briefly, ovarian S10 fractions (40 g protein) were added to assay mixture containing 50 mM glycine, pH 8.3, 20% glycerol, 2 mM DTT, 150 mM KCl, 10 mM MgCl2, 0.1 mg/ml BSA, 0.1 mM NAD+, 20 ng of recombinant eIF5A protein and 2.0 µCi of [3H]-spermidine in a final volume of 25 µl. The reaction mixture was incubated at 37°C for 2h and terminated by the addition of 5µL of SDS-PAGE sample buffer. The proteins were separated on a 12% SDS-PAGE gel, transferred to nitro-cellulose membrane and subjected to flurography using Kodak BioMax TranScreen LE and BioMax MS film at −70°C for 72 hours. The images were then scanned and quantitated.
Gulappa T., Menon B, & Menon K.M. (2015). Hypusination of Eukaryotic Initiation Factor 5A via cAMP-PKA-ERK1/2 Pathway is required for Ligand-induced Downregulation of LH receptor mRNA Expression in the Ovary. Molecular and cellular endocrinology, 413, 90-95.
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6

SidJ Glutamylase Activity Assay

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0.1 μg His6-SdeA, 1 μg GST-SidJ were incubated in a 25 μL reaction system containing 50 mM Tris-HCl (pH 7.5) and 1 mM DTT, 5 mM MgCl2, 1 mM L-glutamate. 1 mM ATP and 1 μM CaM for 2 h at 37°C. To measure the glutamylase activity of SidJ using 14C-glutamate, 2 μg His6-SdeA and 0.5 μg GST-SidJ were incubated in a 25 μL reaction system containing 50 mM Tris-HCl (pH 7.5), 1 mM DTT, 5 mM MgCl2, 1 μCi 14C-L-glutamate (Perkin Elmer cat# NEC290E050UC), 1 mM ATP and 1 μM CaM for 2 h at 37°C. Products were resolved by SDS-PAGE gel and stained with Coomassie brilliant blue. Gels were then dried and signals were detected with x-ray films with a BioMax TranScreen LE (Kodak) for 3 d in −80°C.
Gan N., Zhen X., Liu Y., Xu X., He C., Qiu J., Liu Y., Fujimoto G.M., Nakayasu E.S., Zhou B., Zhao L., Puvar K., Das C., Ouyang S, & Luo Z.Q. (2019). Regulation of phosphoribosyl ubiquitination by a calmodulin-dependent glutamylase. Nature, 572(7769), 387-391.
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