Dsred rab5

Manufactured by Addgene

DsRed-Rab5 is a fluorescent protein construct that consists of the DsRed protein fused to the Rab5 protein. Rab5 is a small GTPase that regulates early endosome fusion and trafficking. The DsRed fluorescent protein provides a red fluorescent signal that can be used to visualize the localization and dynamics of Rab5 within cells.

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Lab products found in correlation

Dsred rab7, by Addgene (4 mentions) Lipofectamine 2000, by Thermo Fisher Scientific (3 mentions) Flag rubicon, by Addgene (2 mentions) Pepstatin a, by Merck Group (1 mentions) Ecl western blotting detection reagent, by Thermo Fisher Scientific (1 mentions) Streptavidin beads, by Thermo Fisher Scientific (1 mentions) Ammonium chloride, by Thermo Fisher Scientific (1 mentions) Polyethylenimine (pei), by Polysciences (1 mentions) Lamp1 rfp, by Addgene (1 mentions) Pierce universal nuclease, by Thermo Fisher Scientific (1 mentions) Protease inhibitor cocktail, by Merck Group (1 mentions) Mcherry rab11, by Addgene (1 mentions)

4 protocols using dsred rab5

Plasmid Transfection in Cells

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DsRed-Rab5, DsRed-Rab7 and Flag-Rubicon plasmids were from Addgene. DsRed-LC3 plasmids were prepared in our laboratory. KSHV Flag-vBcl-2 plasmid was kindly provided by Professor Beth Levine, University of Texas Southwestern Medical Center. All plasmid constructs were confirmed by DNA sequencing. Cells were transiently transfected with the plasmids using lipofectamine 2000 (Invitrogen, Carlsbad, CA, USA) according to the manufacturer's instructions.

Zhang X., Yang Y., Liang X., Zeng X., Liu Z., Tao W., Xiao X., Chen H., Huang L, & Mei L. (2014). Enhancing Therapeutic Effects of Docetaxel-Loaded Dendritic Copolymer Nanoparticles by Co-Treatment with Autophagy Inhibitor on Breast Cancer. Theranostics, 4(11), 1085-1095.

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Reagents and Antibodies for Cell Biology

Cited 1 time

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The following reagents
and antibodies were purchased from commercial sources: Antibodies
against β-actin HRP (sc-4777), CTSL (sc-32320), GAPDH (sc-47724
HRP), normal mouse IgG (sc-2025), normal rabbit IgG (sc-2027) along
with Protein A/G PLUS-Agarose beads (sc-2003) were purchased from
Santa Cruz. Antibodies against LC3 (Cat. #12741) and Streptavidin-HRP
(Cat. #3999) were purchased from Cell Signaling Technology. Antibodies
against PSAP (A1819) and NPC2 (A5413) were purchased from Abclonal.
Protease inhibitor cocktail was purchased from Sigma-Aldrich (Cat.
P8340). Streptavidin beads, ECL Western blotting detection reagent,
and Pierce Universal nuclease were purchased from ThermoScientific.
ClarityMax Western blotting detection reagent was purchased from BioRad
(Cat. 1705062). Polyethylenimine (PEI) was purchased from Polysciences
(Cat. 4765). Inhibitors used were all purchased as follows: Bafilomycin
from CST (Cat. 54645), chloroquine diphosphate from TCI (C2301), ammonium
chloride from Fisher Scientific (A661), E64d from SeleckChem (S7393),
and Pepstatin A from Sigma (P5318). BCA assay was used for protein
concentrations. Expression plasmids for organelle markers were all
purchased from Addgene: mCherry-Sec61 (#49155), dsRed-Rab5 (#13050),
mCherry-Rab11 (#55124), dsRed-Rab7 (#12661), and Lamp1-RFP (#1817).^{40 (link)−43 (link)}

Smith M.R., Zhang L., Jin Y., Yang M., Bade A., Gillis K.D., Jana S., Bypaneni R.N., Glass T.E, & Lin H. (2023). A Turn-On Fluorescent Amino Acid Sensor Reveals Chloroquine’s Effect on Cellular Amino Acids via Inhibiting Cathepsin L. ACS Central Science, 9(5), 980-991.

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Fluorescent Plasmid-based Autophagy Assay

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The EGFP-LC3 and DsRed-LC3 plasmids were created in our laboratory. The DsRed-Rab5, DsRed-Rab7 and Flag-Rubicon plasmids were purchased from Addgene. The KSHV Flag-vBcl-2 plasmid was kindly provided by Professor Beth Levine from the University of Texas Department of Medicine. The Rab1-37 genes in the T Vector were from Professor Jiahuai Han's Lab and sub-cloned into EGFP-C1 and DsRed-C1, respectively. All plasmids were confirmed by automated DNA sequencing. Cells were transiently transfected with the plasmids using Lipofectamine 2000 (Invitrogen) according to the manufacturer's instructions.

Zhang J., Zhang X., Liu G., Chang D., Liang X., Zhu X., Tao W, & Mei L. (2016). Intracellular Trafficking Network of Protein Nanocapsules: Endocytosis, Exocytosis and Autophagy. Theranostics, 6(12), 2099-2113.

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Dual-Fluorescent Protein Plasmid Transfection

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DsRed-Rab5 and DsRed-Rab7 were obtained from Addgene. KSHV Flag-vBcl-2 plasmid was kindly provided by Professor Beth Levine from Department of Medicine, University of Texas. Rab family genes in the T Vector and sub-cloned into EGFP-C1 and DsRed-C1 were kindly provided by Professor Jiahuai Han’s Lab. All the plasmids were confirmed by automated DNA sequencing. And cells were transfected with the plasmids by Lipofectamine 2000 (Invitrogen).

Sun X., Cheng C., Zhang J., Jin X., Sun S., Mei L, & Huang L. (2019). Intracellular Trafficking Network and Autophagy of PHBHHx Nanoparticles and their Implications for Drug Delivery. Scientific Reports, 9, 9585.

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