Sc 108080

Manufactured by Santa Cruz Biotechnology

Sourced in United States, China

Sc-108080 is a laboratory tool designed for scientific research purposes. It serves as a general-purpose device for various experimental applications. Further details about its core function and intended use are not available at this time.

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Lab products found in correlation

Close spelling variants of this product

Control shRNA lentiviral particles (sc-108080) (9 citations) Sc-108080-V (7 citations) Control shRNA lentiviral particles-A (sc-108080) (7 citations) Control shRNA (sc-108080) (5 citations) Control shRNA lentivirus (sc-108080) (4 citations) Control (sc‑108080) short hairpin RNA (2 citations) LV-control-sh; sc-108080 (2 citations) Scrambled shRNA control (sc-108080) lentiviral particles (2 citations) Control (sc‐108080) short hairpin RNA (shRNA) lentiviral particles (2 citations)

144 protocols using sc 108080

Stable Knockdown of Proteins in Huh7 Cells

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We generated stably transduced Huh7 cells with lentivirus vectors encoding shRNA targeting FBP1, p53, or BCCIP (Santa Cruz, CA) or with empty vector alone (SC-108080) following the manufacturer’s protocol. As a control, Huh7 cells were infected with control lentiviral particles with empty vector (Santa Cruz, SC-108080). Stable clones were selected after several passages via puromycin selection in DMEM medium containing three μg/mL of puromycin (Santa Cruz, CA).Stable knockdown of expression of targeted protein was confirmed by Western blot analysis as compared to cells transformed with a vector alone.

Dixit U., Liu Z., Pandey A.K., Kothari R, & Pandey V.N. (2014). Fuse binding protein antagonizes the transcription activity of tumor suppressor protein p53. BMC Cancer, 14, 925.

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Stable Knockdown of AMPK and Nrf2 in SH-SY5Y Cells

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Lentiviral particles, encoding AMPKα1 short hairpin RNA (shRNA;sc-44281-V), Nrf2 shRNA (sc-44332-V) or control shRNA (sc-108080) were purchased from Santa Cruz Biotechnology. SH-SY5Y cells were plated onto six-well plates at a density of 2 × 10⁵ cells/mL in the polybrene-containing complete medium. Lentivirus particles were added to SH-SY5Y cells for 12h. Puromycin (3.0 μg/mL) was then added to select stable cells for a total of six passages (10-12 days). AMPKα1or Nrf2 silencing in stable cells was tested by western blotting.

Mo Y., Zhu J.L., Jiang A., Zhao J., Ye L, & Han B. (2019). Compound 13 activates AMPK-Nrf2 signaling to protect neuronal cells from oxygen glucose deprivation-reoxygenation. Aging (Albany NY), 11(24), 12032-12042.

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Detailed Molecular Signaling Pathway Protocol

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FGF9 (catalogue # 273-F9-025) was purchased from R&D Systems. METAFECTENE® PRO (#T040) was purchased from Biontex. TRPA1 antibodies were purchased from Alomone (#ACC-037) and Santa-Cruz (#sc-166469). Total FGFR2 antibody (#sc-122) was purchased from Santa-Cruz. pFGFR2 (#3471), pERK (#4370), Total ERK (#9102) β-actin (#4970), Flag (#8146), Ki-67 (#9449), β-tubulin (#2146), Zona Occludin-1 (#5406), Claudin-5 (#4933), IgG antibody (#2729), anti-GFP (#2956) antibodies along with U0126 (#9903) were purchased from Cell Signalling. U73122 (#1268) was purchased from TOCRIS. NEB 5-alpha competent E.Coli (high efficiency) (#C2987H) cells purchased from New England Biolabs, were used for transformation. TRPA1 inhibitor HC030031 (#2896) was purchased from Tocris bioscience. ATP (#9804) and kinase buffer (#9802) were purchased from Cell Signalling. Occludin (H-279) (sc-5562) antibody, validated shRNAs^{18 (link), 55 (link)} (sc-29218-V, sc-108080, and sc-29452-V), and DMA (sc-202459) were purchased from Santa Cruz Biotechnology, Inc. Strep antibody (# ab76949) was bought from abcam. CD-63 antibody (#GTX41877) from GeneTex was utilized as an exosomal marker (exosomal proteins were extracted using the Total Exosome RNA and Protein Isolation kit (#4478545, Invitrogen). All antibodies were used in 1:1000 dilutions for western blots.

Berrout J., Kyriakopoulou E., Moparthi L., Hogea A.S., Berrout L., Ivan C., Lorger M., Boyle J., Peers C., Muench S., Gomez J.E., Hu X., Hurst C., Hall T., Umamaheswaran S., Wesley L., Gagea M., Shires M., Manfield I., Knowles M.A., Davies S., Suhling K., Gonzalez Y.T., Carragher N., Macleod K., Abbott N.J., Calin G.A., Gamper N., Zygmunt P.M, & Timsah Z. (2017). TRPA1–FGFR2 binding event is a regulatory oncogenic driver modulated by miRNA-142-3p. Nature Communications, 8, 947.

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Genetically Engineered Spred2 Constructs

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Myc-tagged Spred2 and deletion mutants (ΔEVH1, ΔKBD and ΔSPR) were kindly provided by Prof. John K Heath. Mutations for tryptophan 370/leucine 373 and tyrosine 386/leucine 389 were mutated to Alanine in Spred2 and introduced by PCR-based site-directed mutagenesis. The resulting construct was named Spred2-AA. The GFP-tagged Spred2 was constructed per standard molecular cloning procedures. FLAG-tagged p62 was kindly provided by Prof. Haining Zhu. mRFP-GFP-tagged LC3 was kindly provided by Yoshimori [43 (link)]. GFP-tagged LC3 was purchased from Addgene. Adenoviruses expressing Myc-tagged Spred2 and Myc-tagged Spred2-ΔSPR and the control virus (Ad-Vector) were previously described [8 (link)]. The AdEasy XL adenoviral vector system (Stratagene) was used to generate adenovirus-Myc-Spred2-AA. Primers used were as follows: 5′-GGGGTACCATGACCGAAGAAACACACC-3′ and 5′-GCCAAGCTTATGGTGATGGTGATGATG-3′. The following lentiviral constructs were purchased from Santa Cruz: p62 (SQSTM1) shRNA (sc-29679-V), ATG5 shRNA (sc-41445-V), MAP LC3β shRNA (sc-43390-V) and non-coding shRNA (sc-108080). Lentiviral particles were used to directly infect 293T, A549 and HeLa cells. Stable clones were then selected using puromycin (Sigma). The selected cell populations were subjected to immunoblotting to investigate the silencing efficiency.

Jiang K., Liu M., Lin G., Mao B., Cheng W., Liu H., Gal J., Zhu H., Yuan Z., Deng W., Liu Q., Gong P., Bi X, & Meng S. (2016). Tumor suppressor Spred2 interaction with LC3 promotes autophagosome maturation and induces autophagy-dependent cell death. Oncotarget, 7(18), 25652-25667.

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Modulating KRAS Expression in Cells

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KRAS was silenced via lentiviral transduction of human KRAS shRNA (SC-35731-V; Santa Cruz Biotechnology), and mouse Kras shRNA (iV048022; abm Inc.). Scramble shRNA control (SC-108080; Santa Cruz Biotechnology) and GFP (sc-108084, Santa Cruz Biotechnology) constructs were also used. Maximal knockdown occurred 72 to 96 h after transduction. KRAS^G12V and KRAS^WT were overexpressed using KRAS^G12V lentiviral activation particles (LVP1139-GP; GenTarget Inc.) and KRAS^WT lentiviral activation particles (LVP201104; abm Inc), respectively, following the manufacturer’s protocol. Control lentiviral activation particles (sc-437282; Santa Cruz Biotechnology) served as controls.

Yoon C., Till J., Cho S.J., Chang K.K., Lin J.X., Huang C.M., Ryeom S, & Yoon S.S. (2019). KRAS activation in gastric adenocarcinoma stimulates epithelial-to-mesenchymal transition to cancer stem-like cells and promotes metastasis. Molecular cancer research : MCR, 17(9), 1945-1957.

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Lentiviral Nrf2 Knockdown Protocol

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Two different lentivirus-packed Nrf2 shRNAs, targeting non-overlapping sequence of human Nrf2 (sc-37030-V/“shNrf2–1” and sc-44332-V/“shNrf2–2”), as well as the lentiviral murine Nrf2 shRNA [sc-37049-V, “shNrf2 (m)”] and the scramble nonsense control shRNA (“shC”, sc-108080) were purchased from Santa Cruz Biotech (Santa Cruz, CA). shRNA lentivirus were added to cultured cells in the presence of polybrene (5 μg/mL) for 48 h. Puromycin (1.0 μg/mL) was then included to select stable cells for 4–5 passages. Nrf2 knockdown in the stable cells was confirmed by Western blotting assay and qPCR assay.

Liu H., Feng Y., Xu M., Yang J., Wang Z, & Di G. (2018). Four-octyl itaconate activates Keap1-Nrf2 signaling to protect neuronal cells from hydrogen peroxide. Cell Communication and Signaling : CCS, 16, 81.

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Silencing CHMP5 in Jurkat T cells

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Jurkat cells were grown in RPMI 1640 media supplemented with 10% fetal bovine serum (Sigma-Aldrich, St Louis, MO, USA), 50 U ml⁻¹ penicillin and 50 μg ml⁻¹ streptomycin at 37 °C in an atmosphere of 5% CO₂/95% air. Jurkat T cells were infected with control shRNA lentiviral particles (sc-108080, Santa Cruz Biotechnology, Santa Cruz, CA, USA) or CHMP5 shRNA (h) lentiviral particles (sc-60374-V, Santa Cruz) and selected based on the manufacturer's protocols. Control (Ctrl) Jurkat and CHMP5^KD Jurkat cells were maintained and grown in RPMI 1640 media supplemented with 10% fetal bovine serum (Sigma-Aldrich), 50 U ml⁻¹ penicillin, 4–8 μg ml⁻¹ puromycin and 50 μg ml⁻¹ streptomycin at 37 °C in an atmosphere of 5% CO₂/95% air. The antibodies used were anti-CHMP5 (Abcam, Cambridge, CO, USA), anti-GAPDH (Santa Cruz Biotechnology), anti-TCRαβ (BD Biosciences, San Jose, CA, USA), anti-CD3 (BioLegend, San Diego, CA, USA), anti-CD28 (BioLegend), anti-TCRβ (Abcam), anti-pho-PKCθ (Cell Signaling Technology, Danvers, MA, USA), anti-PKCθ (Cell Signaling Technology), anti-pho-IKKαβ (Cell Signaling Technology), anti-IKKα (Cell Signaling Technology), anti-pho-ZAP-70 (Santa Cruz Biotechnology), anti-ZAP-70 (Santa Cruz Biotechnology), anti-pho-Lck (Santa Cruz Biotechnology) and anti-Lck (Santa Cruz Biotechnology).

Wi S.M., Min Y, & Lee K.Y. (2016). Charged MVB protein 5 is involved in T-cell receptor signaling. Experimental & Molecular Medicine, 48(1), e206-.

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Lentivirus-Mediated Plk1 Knockdown in HASM Cells

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We used lentivirus-mediated shRNA [14 (link)–16 (link), 20 (link)] to generate stable Plk1 knockdown (KD) cells. Briefly, lentiviral particles encoding Plk1 shRNA (sc-36277-V) or control shRNA (sc-108080) were purchased from Santa Cruz Biotechnology. HASM cells were infected with control shRNA lentiviruses or Plk1 shRNA lentiviruses for 12 h. They were then cultured for 3–4 days. Positive clones expressing shRNAs were selected by puromycin. Immunoblot analysis was used to determine the protein levels of Plk1 in these cells. Plk1 KD cells and cells expressing control shRNA were stable at least five passages after initial infection.

Jiang S, & Tang D.D. (2015). Plk1 regulates MEK1/2 and proliferation in airway smooth muscle cells. Respiratory Research, 16(1), 93.

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Lentiviral Knockdown of α7nAChR

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Human α7nAChR shRNA lentiviral particles (sc-42532-V) and control shRNA lentiviral particles (sc-108080) were purchased from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA, USA). The shRNA transfection was performed according to the protocol supplied by the manufacturer. Each milliliter of medium contained 5×10⁴ infectious units of virus. Cells with stable integration of shRNA were selected to be cultured continuously with 10 µg/ml puromycin.

Zhang C., Yu P., Zhu L., Zhao Q., Lu X, & Bo S. (2017). Blockade of α7 nicotinic acetylcholine receptors inhibit nicotine-induced tumor growth and vimentin expression in non-small cell lung cancer through MEK/ERK signaling way. Oncology Reports, 38(6), 3309-3318.

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SMYD3 Knockdown in Prostate Cancer

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SMYD3 knockdown was performed through viral transduction in LNCaP and PC3 cell lines using shRNA Lentiviral Particles (sc-61576-V; Santa Cruz Biotechnology Inc., Santa Cruz, CA) in the presence of polybrene (Santa Cruz Biotechnology Inc.) as described by the manufacturer. Additionally, control LNCaP and PC3 cells were generated using a non-target scramble shRNA (sc-108080; Santa Cruz Biotechnology Inc.). After transduction, stable clones with shRNA were selected with Puromycin dihydrochloride (cat. 631306, Clontech Laboratories Inc.) at a final concentration of 2 μg/ml or 4 μg/ml in LNCaP or PC3 cells, respectively.

Vieira F.Q., Costa-Pinheiro P., Almeida-Rios D., Graça I., Monteiro-Reis S., Simões-Sousa S., Carneiro I., Sousa E.J., Godinho M.I., Baltazar F., Henrique R, & Jerónimo C. (2015). SMYD3 contributes to a more aggressive phenotype of prostate cancer and targets Cyclin D2 through H4K20me3. Oncotarget, 6(15), 13644-13657.

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