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Ad gal

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Ad-gal is a recombinant adenovirus vector that expresses the bacterial beta-galactosidase (lacZ) gene. It can be used for gene delivery and expression studies in various cell types.

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Lab products found in correlation

Ad capn1, by SignaGen (3 mentions) Pcdna3 egfp, by Addgene (1 mentions) Jetprime dna transfection reagent, by Polyplus Transfection (1 mentions) Transmessenger transfection reagent, by Qiagen (1 mentions) H9c2 cell, by American Type Culture Collection (1 mentions) Huvecs, by Lonza (1 mentions) Endothelial cell medium (ecm), by ScienCell (1 mentions)

Spelling variants of this product

Ad-CMV-b-Gal (4 citations) Ad-β-Gal (2 citations)

9 protocols using ad gal

1

Cardiac Microvascular Endothelial Cell Manipulation

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Cardiac microvascular endothelial cells (CMVECs) (within six passages) (CELLutions Biosystems, Toronto, ON, Canada) were infected with adenoviral vector containing rat calpastatin ([Ad-CAST] University of Buffalo, Buffalo, NW, USA) or β-galactosidase ([Ad-gal] Vector Biolabs) at a multiplicity of infection of 100 plaque-forming units/cell. All experiments were performed after 24 h of adenoviral infection.
CMVECs were transfected with pcDNA3-eGFP/β-catenin (Addgene), pcDNA3-eGFP, β-catenin CRISPR/Cas9 knockout (KO) plasmid (β-catenin Double Nickase Plasmid) or a control CRISPR/Cas9 KO plasmid from Santa Cruz Biotechnology, Dallas, TX, USA, using the jetPRIME DNA transfection reagent (Polyplus-Transfection, Illkirch, France) according to the manufacturer’s instructions.
Teng X., Ji C., Zhong H., Zheng D., Ni R., Hill D.J., Xiong S., Fan G.C., Greer P.A., Shen Z, & Peng T. (2019). Selective deletion of endothelial cell calpain in mice reduces diabetic cardiomyopathy by improving angiogenesis. Diabetologia, 62(5), 860-872.
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2

Adenovirus-Mediated Calpain Gene Transfer

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Cultured PMECs were infected with recombinant adenoviruses containing mouse capn1 (Ad-capn1, SignaGen Laboratories), capn2 (Ad-capn2, Applied Biological Materials Inc.), or beta-gal (Ad-gal, Vector Bio-labs) as a control at a multiplicity of infection of 100 PFU/cell. Adenovirus-mediated gene transfer was implemented as previously described [29 (link)].
Liu Z.F., Zheng D., Fan G.C., Peng T, & Su L. (2016). Heat stress prevents lipopolysaccharide-induced apoptosis in pulmonary microvascular endothelial cells by blocking calpain/p38 MAPK signalling. Apoptosis : an international journal on programmed cell death, 21(8), 896-904.
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3

Cardiomyocyte Gene Modulation

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Cultured neonatal cardiomyocytes were infected with adenoviral vectors containing capn1, capn2, pten and mkp-1 gene (Ad-Capn2, Ad-Capn1, Ad-Pten, Ad-MKP-1, SignaGen, USA) or β-gal (Ad-gal, Vector Biolabs, USA) as a control at a multiplicity of infection of 100 plaque forming units/cell (Peng et al. 2003 (link)).
Zheng D., Su Z., Zhang Y., Ni R., Fan G.C., Robbins J., Song L.S., Li J, & Peng T. (2019). Calpain-2 promotes MKP-1 expression protecting cardiomyocytes in both in vitro and in vivo mouse models of doxorubicin-induced cardiotoxicity. Archives of toxicology, 93(4), 1051-1065.
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4

Calpastatin Overexpression in HUVECs

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Human umbilical vein endothelial cells (HUVECs) were isolated from umbilical cord veins and cultured as we previously described [29 (link)]. The isolation of the HUVECs was performed conforming the declaration of Helsinki and approved by the ethics review board at the University of Western Ontario. Cells at passage 1–5 were used in this study.
HUVECs were infected with adenoviral vectors containing rat calpastatin gene (Ad-CAST, University of Buffalo, USA) or beta-gal (Ad-gal, Vector Biolabs) as a control at a multiplicity of infection (MOI) of 10 PFU/cell. Adenovirus-mediated gene transfer was implemented as previously described [30 (link)]. All experiments were performed after 24 hours of adenoviral infection.
Chen B., Zhao Q., Ni R., Tang F., Shan L., Cepinskas I., Cepinskas G., Wang W., Schiller P.W, & Peng T. (2014). Inhibition of calpain reduces oxidative stress and attenuates endothelial dysfunction in diabetes. Cardiovascular Diabetology, 13, 88.
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5

Adenovirus-Mediated ABAT Overexpression

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For in vitro ABAT over-expression, neonatal cardiomyocytes were infected with an adenoviral vector containing human Abat (Ad-ABAT, Vector Biolabs, Philadelphia, PA, USA) at a multiplicity of infection of 100 PFU/cell. An adenoviral vector containing beta-gal (Ad-gal, Vector Biolabs, Philadelphia, PA, USA) served as a control. Adenovirus-mediated gene transfer was implemented as previously described [14 (link)].
Zhang M., Zhong H., Cao T., Huang Y., Ji X., Fan G.C, & Peng T. (2022). Gamma-Aminobutyrate Transaminase Protects against Lipid Overload-Triggered Cardiac Injury in Mice. International Journal of Molecular Sciences, 23(4), 2182.
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6

Adenoviral miR-195 Overexpression in Cardiomyocytes

Cited 1 time
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Cardiomyocytes and endothelial cells were infected with adenoviral vectors containing miR-195 (Ad-miR-195; Vigene Biosciences, Rockville, MD, USA) or β-gal (Ad-gal; Vector Biolabs, Philadelphia, PA, USA) as a control at a multiplicity of infection (MOI) of 50–100 PFU/cell, as we previously described [28 (link)].
Zheng D., Ma J., Yu Y., Li M., Ni R., Wang G., Chen R., Li J., Fan G.C., Lacefield J.C, & Peng T. (2015). Silencing of miR-195 reduces diabetic cardiomyopathy in C57BL/6 mice. Diabetologia, 58(8), 1949-1958.
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7

Modulating Calpain Signaling in Cardiac Cells

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The neonatal mouse cardiomyocytes were prepared and cultured according to methods we described previously [30 (link)].
The rat cardiomyocyte-like H9c2 cells were purchased from the American Type Culture Collection (ATCC) and cultured H9c2 cells were employed within 10 generations.
Cells were infected with adenoviral vectors containing human capn1 gene (Ad-capn1, SignaGen Laboratories), human capn2 gene (Ad-capn2), rat calpastatin gene (Ad-CAST), or beta-gal (Ad-gal, Vector Biolabs) as a control at a multiplicity of infection (MOI) of 100 PFU/cell. Adenovirus-mediated gene transfer was implemented as previously described [10 (link)]. All experiments were performed after 24 h of adenoviral infection.
Cells were transfected with siRNA specific for capn1 and capn2 (Santa Cruz Biotechnology, Inc.) using TransMessenger Transfection Reagent (Qiagen) as we previously described [11 (link)]. A scrambled siRNA served as a control.
Zheng D., Wang G., Li S., Fan G.C, & Peng T. (2015). Calpain-1 induces endoplasmic reticulum stress in promoting cardiomyocyte apoptosis following hypoxia/reoxygenation. Biochimica et biophysica acta, 1852(5), 882-892.
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8

HUVEC Adenoviral Transduction Assay

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Human umbilical vein endothelial cells (HUVECs) were purchased from Lonza (Verviers, Belgium) and cultured in ECM (ScienCell Research Laboratories, USA) containing 10% fetal bovine serum (FBS) at 5% CO2, and 37 °C. Cells were used for experiments up to passage number 5. HUVECs were infected with adenoviral vectors containing rat calpastatin ([Ad-CAST] University of Buffalo, Buffalo, USA) or β-galactosidase ([Ad-gal] Vector Biolabs) at a multiplicity of infection of 100 plaque-forming units/cell. All experiments were performed after 24 h of adenoviral infection.
Yi C., Wu W., Zheng D., Peng G., Huang H., Shen Z, & Teng X. (2020). Targeted inhibition of endothelial calpain delays wound healing by reducing inflammation and angiogenesis. Cell Death & Disease, 11(7), 533.
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9

Adenoviral Vectors for Calpastatin Expression

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Cardiomyocytes were infected with Ad-ATP5A1, adenoviral vectors containing mitochondria-targeted rat calpastatin (Ad-mtCAST; SignaGen Laboratories), or β-gal (Ad-gal; Vector Biolabs, Philadelphia, PA) as a control at a multiplicity of infection of 100 plaque-forming units/cell as previously described (27 (link)).
Ni R., Zheng D., Xiong S., Hill D.J., Sun T., Gardiner R.B., Fan G.C., Lu Y., Abel E.D., Greer P.A, & Peng T. (2015). Mitochondrial Calpain-1 Disrupts ATP Synthase and Induces Superoxide Generation in Type 1 Diabetic Hearts: A Novel Mechanism Contributing to Diabetic Cardiomyopathy. Diabetes, 65(1), 255-268.
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