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F3165 2 mg

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F3165–2 MG is a laboratory equipment product from the Merck Group. It is a standard laboratory instrument designed for general use in research and scientific applications. The core function of this product is to provide a reliable and accurate measurement tool for various laboratory tasks. A detailed description of the intended use or specific applications is not available.

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Lab products found in correlation

Ab6718, by Abcam (2 mentions) Ab15580, by Abcam (2 mentions) R10367, by Abcam (2 mentions) Bz x810 microscope, by Keyence (2 mentions) Vectashield antifade mounting medium, by Vector Laboratories (2 mentions) Normal goat serum (ngs), by Merck Group (2 mentions) Ab191181, by Abcam (2 mentions) Ab22035, by Abcam (2 mentions) Ab5790, by Abcam (2 mentions) H1029 2ml, by Merck Group (1 mentions) Anti flag, by Merck Group (1 mentions) Catalog no arg65350, by Arigo Biolaboratories (1 mentions) Anti p63 4a4 sc 8431, by Santa Cruz Biotechnology (1 mentions) Anti myc catalog no 60003 1, by Proteintech (1 mentions) Anti β actin sc 47778, by Santa Cruz Biotechnology (1 mentions) Amersham imager 600, by GE Healthcare (1 mentions) Ecl plus western blotting detection reagent, by Thermo Fisher Scientific (1 mentions) Aqp11 a, by Alpha Diagnostic (1 mentions) Amersham imager, by Cytiva (1 mentions)

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F3165 (244 citations)

5 protocols using f3165 2 mg

1

Comprehensive Protein Extraction and Analysis

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For protein extraction, seedlings were frozen in liquid nitrogen, ground into powder, then resuspended in 2× SDS buffer (0.125 M Tris-HCl [pH 6.8], 4% SDS, 20% glycerol, 1× cocktail of protease and phosphatase inhibitors, 1 mM PMSF). Samples were heated for 10 min at 65 °C, then centrifuged at 13,000 × g for 10 min at room temperature. The supernatants were transferred into new tubes, and the total protein concentrations were determined by the BCA method. Equal amounts of total proteins were separated in 10% sodium dodecyl sulfate (SDS)-polyacrylamide gels and then transferred onto PVDF membranes. The subsequent immunoblots were performed as previously described52 (link). Antibodies used in this study were anti-HY5 (A gift from Rongcheng Lin’s lab, 1:1000 dilution), anti-BZR1 (A gift from Jianming Li’s lab, 1:1000 dilution), anti-Histone H3 (05-499, Millipore, 1:1000 dilution), anti-HSP (AbM51099-31-PU, Beijing Protein Innovation, 1:5000 dilution), anti-HA (H9658-.2 ML, Sigma-Aldrich, 1:2000 dilution), anti-Flag (F3165-.2MG, Sigma-Aldrich, 1:2000 dilution), Anti-phospho-GSK3 (Tyr279/Tyr216) (05-413, Millipore, 1:1000 dilution), anti-MBP (#E8031S, New England Biolabs, 1:5000 dilution), anti-His (H1029-.2ML, Sigma-Aldrich, 1:2000 dilution), and anti-GST (#2625, Cell Signaling Technology, 1:1000 dilution).
Li J., Terzaghi W., Gong Y., Li C., Ling J.J., Fan Y., Qin N., Gong X., Zhu D, & Deng X.W. (2020). Modulation of BIN2 kinase activity by HY5 controls hypocotyl elongation in the light. Nature Communications, 11, 1592.
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2

Generation of Flag-tagged HBx Plasmid

Cited 1 time
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The Flag-tagged HBx plasmid was generated by inserting the full-length cDNA amplified by PCR into the 2xFlag-pcDNA3 vector between the EcoRI and XhoI sites, using the following primers, 5′-CGGAATTCAATGGCTGCTCGGGTGTGC-3′ and 5′-CCGCTCGAGTTAGGCAGAGGTGAAAAAGTTGC-3′. The plasmids encoding TAp63γ and ΔNp63 were described previously.[22 (link)] The antibodies of anti-Myc (Catalog No. 60003-1, Proteintech, Hubei, China), anti-p21 (Catalog No. #2947, Cell Signaling Technology, Danvers, MA, USA), anti-β-actin (Sc-47778, Santa Cruz, CA, USA), anti-p63 (4A4) (Sc-8431, Santa Cruz), anti-Flag (Sigma-Aldrich, F3165-2MG, St louis, MO, USA), and the secondary antibody for mice (Catalog No. ARG65350, Arigo, Shanghai, China) were commercially purchased.
Xie B., Hao Q., Zhou X, & Chen D. (2022). Inactivation of tumor suppressor TAp63 by hepatitis B virus X protein in hepatocellular carcinoma. Chinese Medical Journal, 135(14), 1728-1733.
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3

Immunostaining of Lentivirus-Transduced Brain Sections

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Lentivirus injected brain sections were post-fixed with 4% PFA for 10 min. Then both lentivirus injected and hNPC injected brain sections were washed with PBS three times and incubated 30 min with blocking solution (PBS/0,1% Triton X-100 containing 10% normal goat serum [Sigma-Aldrich]) and then incubated overnight at 4 °C in blocking solution with primary antibodies: mouse anti-Flag (Sigma, F3165–2 MG, 1:1000); rabbit anti-red fluorescence protein (RFP) (Invitrogen, R10367, 1:1000); rabbit anti-PAX6 (Abcam, ab5790, 1:50); mouse anti-Nestin (Abcam, ab22035, 1:100), rabbit anti-Ki67 (Abcam, ab15580, 1:1000) and mouse anti-human nuclear antigen (Abcam, ab191181, 1:1000). Sections were washed with PBS and incubated for 1 h at room temperature with the secondary antibodies: goat anti-mouse IgG Alexa Fluor 488 (Thermo Fisher, A31560, 1:500); goat anti-rabbit IgG TRITC (Abcam, ab6718, 1:1000) and goat anti-mouse IgG Alexa Fluor 647 (Invitrogen, A32728, 1:1000) diluted in blocking solution. The sections were washed with PBS and mounted with Vectashield Antifade Mounting Medium (Vector Labs, H-1000). Immunofluorescence was visualized and imaged with a Keyence BZ-X810 microscope.
Rufino-Ramos D., Lule S., Mahjoum S., Ughetto S., Bragg D.C., de Almeida L.P., Breakefield X.O, & Breyne K. (2022). Using genetically modified extracellular vesicles as a non-invasive strategy to evaluate brain-specific cargo. Biomaterials, 281, 121366.
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4

Immunostaining of Lentivirus-Transduced Brain Sections

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Lentivirus injected brain sections were post-fixed with 4% PFA for 10 min. Then both lentivirus injected and hNPC injected brain sections were washed with PBS three times and incubated 30 min with blocking solution (PBS/0,1% Triton X-100 containing 10% normal goat serum [Sigma-Aldrich]) and then incubated overnight at 4 °C in blocking solution with primary antibodies: mouse anti-Flag (Sigma, F3165–2 MG, 1:1000); rabbit anti-red fluorescence protein (RFP) (Invitrogen, R10367, 1:1000); rabbit anti-PAX6 (Abcam, ab5790, 1:50); mouse anti-Nestin (Abcam, ab22035, 1:100), rabbit anti-Ki67 (Abcam, ab15580, 1:1000) and mouse anti-human nuclear antigen (Abcam, ab191181, 1:1000). Sections were washed with PBS and incubated for 1 h at room temperature with the secondary antibodies: goat anti-mouse IgG Alexa Fluor 488 (Thermo Fisher, A31560, 1:500); goat anti-rabbit IgG TRITC (Abcam, ab6718, 1:1000) and goat anti-mouse IgG Alexa Fluor 647 (Invitrogen, A32728, 1:1000) diluted in blocking solution. The sections were washed with PBS and mounted with Vectashield Antifade Mounting Medium (Vector Labs, H-1000). Immunofluorescence was visualized and imaged with a Keyence BZ-X810 microscope.
Rufino-Ramos D., Lule S., Mahjoum S., Ughetto S., Bragg D.C., de Almeida L.P., Breakefield X.O, & Breyne K. (2022). Using genetically modified extracellular vesicles as a non-invasive strategy to evaluate brain-specific cargo. Biomaterials, 281, 121366.
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5

Quantifying Aquaporin Expression in Oocytes

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The total and surface biotinylated samples from oocytes AQP injected with cRNA encoding for AQPs or H2O-injected controls, were separated by SDS-PAGE on 12% Tris-glycine gels. The samples were transferred to PVDF membranes and incubated in TBST with 5% of milk for 1 h at room temperature. The membranes were probed overnight at 4°C with polyclonal anti-hAQP1 (AQP11-A, Alpha Diagnostics, San Antonio, USA), anti-rAQP3 (AQP31-A, Alpha Diagnostics), hAQP7 (AQP71-A, Alpha Diagnostics), anti-hAQP8 (AQP81-A, Alpha Diagnostics), anti-hAQP9 (AQP91-A, Alpha Diagnostics) or monoclonal anti-FLAG (F3165-2MG, Sigma Aldrich) antibodies. The protein expression was detected using ECL plus Western Blotting Detection Reagents (32132, Thermo Fisher Scientific) and the signals were detected on an Amersham Imager 600 (GE Healthcare).
Kabutomori J., Beloto-Silva O., Geyer R.R, & Musa-Aziz R. (2018). Lithobates catesbeianus (American Bullfrog) oocytes: a novel heterologous expression system for aquaporins. Biology Open, 7(4), bio031880.
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