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Nb500 169

Manufactured by Thermo Fisher Scientific
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The NB500-169 is a laboratory instrument designed for liquid handling applications. It is a precision pipette that can accurately and reliably transfer small volumes of liquid samples or reagents. The core function of this product is to facilitate fluid transfers in various laboratory workflows.

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Lab products found in correlation

Ma5 14520, by Thermo Fisher Scientific (2 mentions) Evos fl auto 2 inverted fluorescence microscope, by Thermo Fisher Scientific (2 mentions) Imager m2 upright fluorescence microscope, by Zeiss (2 mentions) Fluorescent secondary antibody, by Thermo Fisher Scientific (2 mentions) Gfp 1020, by Aves Labs (2 mentions) Ab5535, by Novus Biologicals (1 mentions) Vectorshield with dapi, by Vector Laboratories (1 mentions) Alexa fluor 488 and 555, by Thermo Fisher Scientific (1 mentions) Anti idu, by BD (1 mentions)

3 protocols using nb500 169

1

Immunocytochemical Analysis of Astrocytes

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We fixed and permeablized astrocytes with 4% paraformaldehyde and 0.2% Triton‐X100 in phosphate buffered saline (PBS). After blocking in 10% donkey serum, we stained astrocytes with the following primary antibodies: anti‐GFAP (1:1,500. Biolegend, 829401), Ki67 (1:200. Invitrogen MA5‐14520), enhanced green fluorescent protein (EGFP) (1:1,000. Aves Labs GFP‐1020), mCherry (1:600. Clontech 632543), Sox9 (1:2,000. Millipore AB5535), Bromodesoxyuridine (BrdU) (1:500, Novusbio NB500‐169), and fluorescent secondary antibodies (Invitrogen). For BrdU staining, cells were pretreated by 2N hydrochloric acid for 20 min before blocking. After three washes in PBS, we mounted the coverslips with VectorShield with DAPI (Vector Labs H1200) and imaged them using an Evos FL Auto 2 inverted fluorescence microscope (Invitrogen) with ×10, and ×20 lenses and the Imager M2 upright fluorescence microscope (Zeiss) with ×10, ×20, and ×40 lenses. We cropped the images with the FIJI and Photoshop softwares.
Li J., Khankan R.R., Caneda C., Godoy M.I., Haney M.S., Krawczyk M.C., Bassik M.C., Sloan S.A, & Zhang Y. (2019). Astrocyte‐to‐astrocyte contact and a positive feedback loop of growth factor signaling regulate astrocyte maturation. Glia, 67(8), 1571-1597.
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2

Immunofluorescence Staining of Astrocytes

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We fixed and permeablized astrocytes with 4% paraformaldehyde and 0.2% Triton-X100 in phosphate buffered saline (PBS). After blocking in 10% donkey serum, we stained astrocytes with the following primary antibodies: anti-GFAP (1:1500. Biolegend, 829401), Ki67 (1:200. Invitrogen MA5–14520), enhanced green fluorescent protein (EGFP) (1:1000. Aves Labs GFP-1020), mCherry (1:600. Clontech 632543), Sox9 (1:2000. Millipore AB5535), Bromodesoxyuridine (BrdU) (1:500, Novusbio NB500–169), and fluorescent secondary antibodies (Invitrogen). For BrdU staining, cells were pretreated by 2 N hydrochloric acid for 20 minutes before blocking. After three washes in PBS, we mounted the coverslips with VectorShield with DAPI (Vector Labs H1200) and imaged them using an Evos FL Auto 2 inverted fluorescence microscope (Invitrogen) with 10x, and 20x lenses and the Imager M2 upright fluorescence microscope (Zeiss) with 10x, 20x, and 40x lenses. We cropped the images with the FIJI and Photoshop softwares.
Li J., Khankan R.R., Caneda C., Godoy M.I., Haney M.S., Krawczyk M.C., Bassik M.C., Sloan S.A, & Zhang Y. (2019). Astrocyte‐to‐astrocyte contact and a positive feedback loop of growth factor signaling regulate astrocyte maturation. Glia, 67(8), 1571-1597.
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3

DNA Fiber Labeling and Imaging

Cited 1 time
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Cells were labeled with 50 μM CldU (Sigma), exposed to HU (4 mM) and labeled with 50 μM IdU (Sigma) as indicated in the figures. DNA fibers were spread as previously described (35 (link)), and fiber tracts were detected using anti-IdU (BD Biosciences, 347580) and anti-CldU (Novus Biologicals, NB500-169) primary antibodies and Alexa Fluor 488 and 555 secondary antibodies (Invitrogen). Fibers were imaged at 60× magnification with oil immersion using a Zeiss microscope and analyzed with ImageJ.
Eckelmann B.J., Bacolla A., Wang H., Ye Z., Guerrero E.N., Jiang W., El-Zein R., Hegde M.L., Tomkinson A.E., Tainer J.A, & Mitra S. (2020). XRCC1 promotes replication restart, nascent fork degradation and mutagenic DNA repair in BRCA2-deficient cells. NAR Cancer, 2(3), zcaa013.
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