LNCaP (clone FGC) (RRID:CVCL_1379), VCaP (RRID:CVCL_2235), PC3 (RRID:CVCL_0035), RWPE-1 (RRID:CVCL_3791), HEK293T (RRID:CVCL_0063), DU145 (RRID:CVCL_0105), 22Rv1 (RRID:CVCL_1045) and C4–2 (RRID:CVCL_4782) cells were purchased from American Type Culture Collection (ATCC, Manassas, VA). HEK293T and VCaP cells were cultured in DMEM media supplemented with 10% fetal bovine serum, 100 units/ml penicillin and 100 g/ml streptomycin, and 2 mM L-alanyl-L-glutamine (Corning Glutagro™) and 1: 10000 plasmocin (Invivogen, San Diego, CA). LNCaP and 22Rv1 cells were cultured in RPMI 1640 and PC3 cells were cultured in F12K media with the same supplements as above. DU145 and C4–2 cells were cultured based on ATCC suggestions. RWPE-1 was cultured in Keratinocyte SFM (Gibco 17005–042, Thermo Scientific). All other media and supplements were obtained from Corning Mediatech, Inc. For androgen deprivation studies, charcoal-dextran stripped FBS was used at 10% with phenol red-free DMEM or RPMI 1640. All cultures were used up to passage number 20 and tested monthly for Mycoplasma contamination with MycoAlertTM Mycoplasma Detection Kit (Lonza, Switzerland).
L alanyl l glutamine glutagro
Manufactured by Corning
Sourced in United States
L-alanyl-L-glutamine (Corning Glutagro™) is a dipeptide compound composed of the amino acids L-alanine and L-glutamine. It is a cell culture supplement designed to provide a stable source of glutamine for cell growth and proliferation.
Automatically generated - may contain errors
Lab products found in correlation
Plasmocin, by InvivoGen (2 mentions)
Hek293t, by American Type Culture Collection (2 mentions)
Fetal bovine serum (fbs), by Atlanta Biologicals (2 mentions)
Dmem media, by Corning (1 mentions)
Du145, by American Type Culture Collection (1 mentions)
Rwpe 1, by Thermo Fisher Scientific (1 mentions)
Rwpe 1, by American Type Culture Collection (1 mentions)
Keratinocyte sfm, by Thermo Fisher Scientific (1 mentions)
Lncap clone fgc, by American Type Culture Collection (1 mentions)
Hek293t, by Corning (1 mentions)
Mycoalert mycoplasma detection kit, by Lonza (1 mentions)
Mccoy s 5a medium, by Corning (1 mentions)
Mycoalert, by Lonza (1 mentions)
L glutamine, by Corning (1 mentions)
Sodium pyruvate, by Corning (1 mentions)
Fetal bovine serum (fbs), by Corning (1 mentions)
Streptomycin, by Corning (1 mentions)
Penicillin, by Corning (1 mentions)
Minimum essential medium nonessential amino acids, by Corning (1 mentions)
1640 medium, by Corning (1 mentions)
4 protocols using l alanyl l glutamine glutagro
1
Comprehensive Characterization of Prostate Cell Lines
2
Cell Culture and Maintenance Protocols
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All cells were purchased from American Type Culture Collection (ATCC) and only used if cultured for fewer than 30 passages. All cells were screened for mycoplasma (Lonza, MycoAlert) on a weekly basis. Caki-2 cells (ATCC) were cultured in McCoy’s 5A medium (Corning Mediatech, Inc) supplemented with 10% fetal bovine serum (Corning Mediatech, Inc), 1% Minimum Essential Medium nonessential amino acids (Corning Mediatech, Inc), 1% antibiotics (100 units/ml penicillin and 100 μg/ml streptomycin (Corning Mediatech, Inc), 2 mM L-alanyl-L-glutamine (Corning glutagro; Corning Mediatech, Inc), and 2.5 μg/ml plasmocin (InvivoGen, Inc). HEK-293T (ATCC) were cultured in DMEM (Corning Mediatech, Inc) supplemented with 10% fetal bovine serum (Corning Mediatech, Inc), 1% antibiotics (100 units/ml penicillin and 100 μg/ml streptomycin; Corning Mediatech, Inc), 1% sodium pyruvate (Corning Mediatech, Inc), 1% L-glutamine (Corning Mediatech, Inc), and 2.5 μg/ml plasmocin (InvivoGen, Inc). All cells were grown in a humidified atmosphere in a 5% CO2 incubator.
3
Optimized Infection of Mammalian Cells with Coxiella burnetii
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Human cervical epithelial cells (HeLa; ATCC CCL-2; ATCC, Manassas, VA, USA) and mouse alveolar macrophages (MH-S; ATCC CRL-2019) were incubated at 37°C in 5% CO2 and maintained in RPMI (Roswell Park Memorial Institute) 1640 medium (Corning, New York, NY, USA) with 10% fetal bovine serum (FBS; Atlanta Biologicals, Norcross, GA, USA) and 2 mM l -alanyl-l -glutamine (Glutagro; Corning). Human embryonic kidney cells (HEK 293T; ATCC CRL-3216) were incubated at 37°C in 5% CO2 and maintained in Dulbecco’s modified Eagle medium (DMEM; Corning) with 10% FBS. All mammalian cell cultures were passaged every 3 days, and no cells older than 20 passages were used in experiments. Coxiella burnetii Nine Mile phase II (NMII; clone 4, RSA 439) expressing mCherry was grown for 28 days in Vero cells, washed with phosphate-buffered saline (PBS), and stored as previously described (27 (link)). For each mammalian cell type, bacterial multiplicity of infection (MOI) was optimized so that no more than one bacterium was internalized in each infected cell.
4
Cultivation and Infection of Coxiella burnetii
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Coxiella burnetii Nine Mile Phase II (NMII) (clone 4, RSA 439), ΔdotA (T4BSS mutant) C. burnetii, and mCherry-expressing wild-type (WT) C. burnetii [27 (link)] were grown for 4 days in ACCM-2, washed twice with phosphate buffered saline (PBS) and stored as previously described [67 (link)]. A mCherry-expressing ΔdotA C. burnetii was generated by electroporating pJB-CAT-1169-mCherry into ΔdotA C. burnetii as described previously [90 (link)]. The multiplicity of infection of each bacteria stock was optimized for each cell type and culture vessel to ~1 internalized bacterium per cell at 37°C and 5% CO2. Human cervical epithelial cells (HeLa, ATCC CCL-2) and mouse alveolar macrophages (MH-S; ATCC CRL-2019) were maintained in RPMI (Roswell Park Memorial Institute) 1640 medium (Corning, New York, NY, USA) containing 10% fetal bovine serum (FBS; Atlanta Biologicals, Norcross, GA, USA) and 2 mM L-alanyl-L-glutamine (glutagro; Cat. 25-015-CI, Corning, New York, NY) at 37°C and 5% CO2. The wild type (parental) and TFEB-GFP expressing HeLa cells (generously provided by Richard J. Youle) [61 (link)] were maintained in DMEM (Dulbecco’s Modified Eagle Medium; Corning) containing 10% FBS at 37°C and 5% CO2.
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