Sigfp

Manufactured by Santa Cruz Biotechnology

Sourced in United States

SiGFP is a protein-based fluorescent marker. It is derived from the green fluorescent protein (GFP) found in the jellyfish Aequorea victoria. SiGFP emits green fluorescence when exposed to blue or ultraviolet light, allowing for the visualization and tracking of cellular and subcellular processes.

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Lab products found in correlation

Si sirt1, by Santa Cruz Biotechnology (1 mentions) Sisirt6, by Santa Cruz Biotechnology (1 mentions) Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (1 mentions) Jetpei, by Polyplus Transfection (1 mentions) Interferin, by Polyplus Transfection (1 mentions)

3 protocols using sigfp

Ectopic Expression of TRF2 Isoforms

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Human cervix carcinoma HeLa cells were purchased by the ATCC repository and maintained according to the manufacturer's instructions. Human immortilized BJ-hTERT and transformed BJ-EHLT/RasV12 fibroblasts were obtained and maintained as described (57 (link),58 (link)). The wild-type and p53-deficient colon carcinoma HCT116 cells were obtained by Dr Vogelstein, Johns Hopkins University. All the cell lines were grown in Dulbecco modified eagle medium (DMEM; Invitrogen, Carlsbad, CA, USA) containing 10% fetal calf serum.
Empty-, TRF2^wt- or TRF2^cT-overexpressing cells were obtained by infecting with amphotrophic retroviruses generated by transient transfection of retroviral vectors (pBabe-puro-Empty, pBabe-puro-mycTRF2 and pLPC-Myc-TRF^cT; the last one was a gift from Eros Lazzerini Denchi, Addgene plasmid #44573; 7 (link)) into Phoenix amphotropic packaging cells with JetPEI (Polyplus, New York, NY, USA), according to the manufacturer's instructions. For transient RNA interference experiments, siTRF2 and siPARP1 were purchased from Dharmacon Inc. (Chicago, USA), siSIRT6, siSIRT1 and siGFP were purchased from Santa Cruz Biotechnology Inc. (Santa Cruz, CA, USA) and transfected into HCT116 or HeLa cells with Interferin (Polyplus) according to the manufacturer's instructions.

Rizzo A., Iachettini S., Salvati E., Zizza P., Maresca C., D'Angelo C., Benarroch-Popivker D., Capolupo A., del Gaudio F., Cosconati S., Di Maro S., Merlino F., Novellino E., Amoreo C.A., Mottolese M., Sperduti I., Gilson E, & Biroccio A. (2016). SIRT6 interacts with TRF2 and promotes its degradation in response to DNA damage. Nucleic Acids Research, 45(4), 1820-1834.

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NS2 Knockdown in A549 Cells

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A549 cells were transduced with Lenti-NS2 or Lenti-NS2^H126R and 48 hr post transduction, medium containing 1 mg/ml of hygromycin was added, after 72 hr selection the cells were plated into a new flask and 24 hr later cloned by limited dilution by previous methods (Li et al., 2016 (link)). For NS2 knockdown, 3 × 10⁵ cells were plated in one well of a 24-well plate, and transfected with 100 pmol siRNA in Lipofectatimine 2000; cells were harvested for western blotting (72 hr post transfection) to determine the expression level of NS2 or fixed with 4% PFA (96 hr post transfection) and stained with crystal violet. siNS2-1 (5' rArUrGrArArGrArUrArUrUrGrUrUrGrArArArUrUrCrUrCCG 3') and siNS2-2 (5' rGrGrArArUrGrCrArArUrUrCrUrArUrCrArUrArArArGrAAC 3') targetting NS2 were designed and synthesized by IDT and the siGFP was purchased from SantaCruz.

Li Y., Banerjee S., Goldstein S.A., Dong B., Gaughan C., Rath S., Donovan J., Korennykh A., Silverman R.H, & Weiss S.R. (2017). Ribonuclease L mediates the cell-lethal phenotype of double-stranded RNA editing enzyme ADAR1 deficiency in a human cell line. eLife, 6, e25687.

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Stable and Transient Transfection of CSN7A/B in Adipogenesis

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For stable transfection, CSN7A or CSN7B cDNA were obtained by PCR and cloned into pCMV‐3Tag‐1a vector (Stratagene, La Jolla, CA, USA) encoding 3 N‐terminal Flag‐tags. These plasmids were transfected into LiSa‐2 cells. Single clones were used to obtain resistant cell lines with 0.25 mg·mL⁻¹ of neomycin.
For transient transfection, human siCSN7A, siCSN7B and siGFP (Cell Signaling, Denver, CO, USA) were transfected in LiSa‐2 cells or mouse siCSN7A, siCSN7B and siGFP (Santa Cruz, Santa Cruz, CA, USA) were transfected in MEF cells. After 48 h, the adipogenic differentiation of LiSa‐2 cells was induced by changing the culture medium (see above). LiSa‐2 cells were then harvested for western blot analysis and for ORO staining at day 1, 8, and 15 of differentiation. For MEFs, adipogenic differentiation was induced by a change in culture medium (see above) for 4 days, after which cells were cultured in normal medium for another 3 days (total of 7 days) 43 and harvested for western blotting and for ORO staining.

Huang X., Ordemann J., Pratschke J, & Dubiel W. (2016). Overexpression of COP9 signalosome subunits, CSN7A and CSN7B, exerts different effects on adipogenic differentiation. FEBS Open Bio, 6(11), 1102-1112.

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