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Bluelf prestained protein ladder

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The BLUelf prestained protein ladder is a molecular weight standard used for determining the approximate size of proteins in gel electrophoresis. It contains a mixture of pre-stained proteins with defined molecular weights, allowing for easy identification of protein bands during the electrophoresis process.

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Lab products found in correlation

Anti mouse igg hrp, by Promega (1 mentions) Supersignal west femto reagent, by Thermo Fisher Scientific (1 mentions) Invitrolon pvdf membrane, by Thermo Fisher Scientific (1 mentions) Pierce ecl plus western blotting substrate, by Thermo Fisher Scientific (1 mentions) Himark pre stained protein standard, by Thermo Fisher Scientific (1 mentions) Gotaq dna polymerase, by Promega (1 mentions) Benzonase nuclease, by Merck Group (1 mentions) Wizard sv gel and pcr clean up system, by Promega (1 mentions) Nitrocellulose membrane, by Bio-Rad (1 mentions) Puromycin, by Thermo Fisher Scientific (1 mentions) Fluo 4, by Thermo Fisher Scientific (1 mentions) Blasticidin s hcl, by Thermo Fisher Scientific (1 mentions) Calcium green 5n, by Thermo Fisher Scientific (1 mentions) Safranine o, by Merck Group (1 mentions) Bca protein assay kit, by Thermo Fisher Scientific (1 mentions) Platinum taq dna polymerase high fidelity, by Thermo Fisher Scientific (1 mentions) Q5 high fidelity dna polymerase, by New England Biolabs (1 mentions) Carbonyl cyanide p trifluoromethoxyphenylhydrazone fccp, by Merck Group (1 mentions) Fluoromount g, by Southern Biotech (1 mentions) T4 dna ligase, by Promega (1 mentions)

5 protocols using bluelf prestained protein ladder

1

Grifolin Compound Protein Expression

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Cells were seeded in 6-well plates at a density of 2.5 x 105 cells/well and treated with grifolin, neogrifolin and confluentin or with 2% DMSO for 24 and 48 hours. Cell lysates were prepared as previously described [23 (link)]. Protein samples were resolved on a 10% SDS-PAGE and transferred onto a nitrocellulose membrane. BLUelf prestained protein ladder (FroggaBio) was used. Anti-mouse IgG-HRP (1:4,000, Promega) was used as secondary antibody. All blots were visualized using chemiluminescent detection system with the FluorChem Q Imager (ProteinSimple, CA, USA) and analyzed using the AlphaView Q software (ProteinSimple).
Yaqoob A., Li W.M., Liu V., Wang C., Mackedenski S., Tackaberry L.E., Massicotte H.B., Egger K.N., Reimer K, & Lee C.H. (2020). Grifolin, neogrifolin and confluentin from the terricolous polypore Albatrellus flettii suppress KRAS expression in human colon cancer cells. PLoS ONE, 15(5), e0231948.
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2

Western Blot Analysis of NBCe1 Protein

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Protein extracts were run on NuPAGE 3%–8% Tris-acetate gels alongside 5 µL of BLUelf prestained protein ladder (FroggaBio, Concord, ON, Canada) or HiMark prestained protein standard (Thermo Fisher Scientific). Blank lanes were loaded with 20 µL NuPAGE LDS sample buffer. Protein bands were transferred onto Invitrolon PVDF membranes (Invitrogen, Carlsbad, CA, USA) and stored in blocking buffer (5% milk powder and 0.1% Tween 20 in TBS) overnight. NBCe1 protein was probed using a 1:1000 dilution of rabbit anti-SLC4A4 polyclonal antibody (#E-AB-14348, Elabscience, Houston, TX, USA; see [38 (link)] and Supplemental Figure S2 for validation) or 1:1000 of our novel rabbit anti-pSer982 antibody (preabsorbed for 30 min with 16 µg/10 mL blocking buffer of peptide (Pep1); see manuscript results for details of validation). Exceptions were the data in Figure 9, for which we incidentally used a different anti-NBCe1-A/B antibody [39 (link)] at a 1:1000 dilution. Immunoreactivity was disclosed using 1:2000 of HRP-conjugated goat-anti-rabbit secondary antibody (MP Biomedicals, Solon, OH, USA) with either Pierce ECL Plus Western Blotting Substrate or SuperSignal West Femto reagent (Thermo Fisher Scientific, Waltham, MA, USA).
Alsufayan T.A., Myers E.J., Quade B.N., Brady C.T., Marshall A., Haque N., Duffey M.E, & Parker M.D. (2021). Revisiting the Role of Ser982 Phosphorylation in Stoichiometry Shift of the Electrogenic Na+/qHCO3− Cotransporter NBCe1. International Journal of Molecular Sciences, 22(23), 12817.
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3

Fluorescent Protein Assays and Molecular Cloning

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Platinum Taq DNA Polymerase High Fidelity, Calcium Green-5N, Fluo-4 and BCA Protein Assay Kit were from Thermo Fisher Scientific (Waltham, MA). BLUelf prestained protein ladder was from FroggaBio (Wheatfield, NY). Puromycin was from Acros Organics (Fair Lawn, NJ). Blasticidin S HCl was purchased from Life Technologies (Grand Island, NY). Wizard® plus SV miniprep Purification System, Wizard® SV Gel and PCR Clean-up System, GoTaq DNA polymerase and T4 DNA ligase were from Promega (Madison, WI). Antarctic phosphatase, restriction enzymes and Q5® High-Fidelity DNA Polymerase were from New England Biolabs (Ipswich, MA). Fluoromount-G® was from SouthernBiotech (Birmingham, AL). The primers were purchased from Integrated DNA Technologies. The Nitrocellulose Membranes was from Bio-Rad (Hercules, CA). G418 sulfate was from KSE Scientific (Durham, NC). Anti-Cas9 rabbit polyclonal antibody (Cat. No. TA190309) was from OriGene (Rockville, MD). Anti-HA mouse monoclonal antibody (Cat. No. 901501) was from BioLegend (San Diego, CA). Anti-tubulin monoclonal antibody (Cat. No. T6199), mammalian cell protease inhibitor mixture (Sigma P8340), other protease inhibitors, safranine O, carbonylcyanide p-trifluoromethoxyphenylhydrazone (FCCP), Benzonase® Nuclease and all other reagents of analytical grade were from Sigma (St. Louis, MO).
Koo E.H., Lansbury PT J.r, & Kelly J.W. (1999). Amyloid diseases: Abnormal protein aggregation in neurodegeneration. Proceedings of the National Academy of Sciences of the United States of America, 96(18), 9989-9990.
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4

Reagents and Materials for Biochemical Assays

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Kanamycin sulfate, isopropyl β-D-1-thiogalactopyranoside (IPTG), 100X MEM vitamin mixture (for bacterial culture work), bovine serum albumin (BSA), Bromophenol Blue, 5-bromouracil (BrU), 5-bromouridine (BrUrd), 3-(trimethylsilyl)-1-propanesulfonic acid-d6 sodium salt (DSS-D6), 5-fluorouracil (FU), 5-fluorouridine (FUrd), nitroblue tetrazolium (NBT), riboflavin, uracil (U) and uridine (Urd) were purchased from Sigma (Oakville, Canada). 5-chlorouracil (ClU) was obtained from Ark Pharm Incorporated (Arlington Heights, USA). The trifluridine (F3TDR) was purchased from Oxchem Corporations (Wood Dale, USA). The BLUelf prestained protein ladder was purchased from FroggaBio (Concord, Canada). The Luria Broth (LB, Invitrogen), Coomassie G-250, HPLC grade H2O and the Spectra/Por 6–8 kDa molecular weight cut-off (MWCO) dialysis tubing were purchased from Fisher Scientific (Ottawa, Canada). The 3.5 kDa MWCO Snakeskin dialysis tubing was purchased from Thermo Fisher Scientific (Whitby, Canada). The 15NH4Cl was purchased from Cambridge Isotope Laboratories (Andover, USA). The MAXYMum Recovery 1.5 mL centrifuge tubes were purchased from Corning (Fairport, USA). The 3 mM SampleJet NMR tubes were obtained from Bruker (Milton, Canada).
LeVatte M., Lipfert M., Roy D., Kovalenko A, & Wishart D.S. (2021). Cloning and high-level expression of monomeric human superoxide dismutase 1 (SOD1) and its interaction with pyrimidine analogs. PLoS ONE, 16(2), e0247684.
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5

SDS-PAGE Protein Separation Protocol

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The cell lysates were pelleted in the microcentrifuge for 10 minutes at 12,000 rpm to spin down debris. Appropriate volumes of each sample were loaded such that all lanes contain the same amount of protein. FroggaBio BLUelf Prestained Protein ladder was used at a volume of 5µl. The gel was run for 45 minutes at 70V (constant voltage) to move the proteins through the stacking gel. It was made sure that all samples go through the stacking gel before increasing the voltage for the resolving gel. The voltage was then changed to 120V and the gel was allowed to run for 2 hours 20 minutes until the tracking dye has begun to leak out into the outer chamber.
Cojocari V. (2021). Assessing the impact of select intestinal tract conditions on expression of lee encoded genes in enterohemorrhagic escherichia.
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