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Nbp2 14833

Manufactured by Novus Biologicals
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NBP2-14833 is a laboratory reagent produced by Novus Biologicals. It serves as a tool for biological research and analysis, but a detailed description of its core function cannot be provided while maintaining an unbiased and factual approach.

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Lab products found in correlation

Lna microrna ish mir 122 optimization kit, by Qiagen (2 mentions) Nuclear fast red, by Vector Laboratories (2 mentions) Sheep anti digoxigenin ap, by Roche (2 mentions) Nbt bcip, by Vector Laboratories (2 mentions) Nb100 449, by Novus Biologicals (1 mentions) 4 hydroxytamoxifen, by Merck Group (1 mentions) Peroxidase goat anti rabbit or mouse h l, by Jackson ImmunoResearch (1 mentions) Sab4200095, by Merck Group (1 mentions)

4 protocols using nbp2 14833

1

Synthesis and Evaluation of TEPP-46 and DASA-58

Cited 1 time
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TEPP-46 and DASA-58 synthesized in accordance with published methods (Boxer et al., 2010 (link); Jiang et al., 2010 (link)). LPS used in vitro (100 ng/ml) and in vivo were E. coli, serotype EH100 (Alexis), and 055:B5 (Sigma-Aldrich). 4-hydroxytamoxifen (H7904) from Sigma. Antibodies: anti-PKM2 (3198), antiphospho-PKM2 (Tyr105), β-actin (4267) (all Cell Signaling Technologies), anti-IL-1β (R&D, AF401-NA), anti-HIF-1α (Novus, NB100-449), and anti-PKM1 (Novus, NBP2-14833). Dimethyl sulfoxide (DMSO) used as vehicle control for TEPP/DASA in all in vitro assays.
Palsson-McDermott E.M., Curtis A.M., Goel G., Lauterbach M.A., Sheedy F.J., Gleeson L.E., van den Bosch M.W., Quinn S.R., Domingo-Fernandez R., Johnson D.G., Jiang J.K., Israelsen W.J., Keane J., Thomas C., Clish C., Heiden M.V., Xavier R.J, & O’Neill L.A. (2015). Pyruvate Kinase M2 regulates Hif-1α activity and IL-1β induction, and is a critical determinant of the Warburg Effect in LPS-activated macrophages. Cell metabolism, 21(1), 65-80.
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2

Immunohistochemistry and In Situ Hybridization Protocols

Cited 1 time
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IHC was performed as previously described69 (link) using a 1:400 antibody dilution for PKM2 (C-11), 1:600 dilution for PKM1 (Cat# NBP2-14833, Novus Biologicals; Littleton, CO), and 1:250 dilution for GLUT1 (AbCam; Cat# ab652). ISH was performed as described70 (link) using LNA™ microRNA ISH miR-122 optimization kit (Cat# 90003, Exiqon; Woburn, MA) followed by incubation of sheep anti-digoxigenin-AP (Cat# 11093274910, Roche Diagnostics; Mannheim, Germany), and developed with NBT:BCIP (Cat# SK-5400, Vector Laboratories; Burlingame, CA) at 30°C overnight. Nuclear fast red was used to counterstain nuclei (Cat# H-3403, Vector Laboratories).
Fong M.Y., Zhou W., Liu L., Alontaga A.Y., Chandra M., Ashby J., Chow A., O’Connor S.T., Li S., Chin A.R., Somlo G., Palomares M., Li Z., Tremblay J.R., Tsuyada A., Sun G., Reid M.A., Wu X., Swiderski P., Ren X., Shi Y., Kong M., Zhong W., Chen Y, & Wang S.E. (2015). Breast cancer-secreted miR-122 reprograms glucose metabolism in pre-metastatic niche to promote metastasis. Nature cell biology, 17(2), 183-194.
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3

Immunohistochemistry and In Situ Hybridization Protocols

Cited 1 time
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IHC was performed as previously described69 (link) using a 1:400 antibody dilution for PKM2 (C-11), 1:600 dilution for PKM1 (Cat# NBP2-14833, Novus Biologicals; Littleton, CO), and 1:250 dilution for GLUT1 (AbCam; Cat# ab652). ISH was performed as described70 (link) using LNA™ microRNA ISH miR-122 optimization kit (Cat# 90003, Exiqon; Woburn, MA) followed by incubation of sheep anti-digoxigenin-AP (Cat# 11093274910, Roche Diagnostics; Mannheim, Germany), and developed with NBT:BCIP (Cat# SK-5400, Vector Laboratories; Burlingame, CA) at 30°C overnight. Nuclear fast red was used to counterstain nuclei (Cat# H-3403, Vector Laboratories).
Fong M.Y., Zhou W., Liu L., Alontaga A.Y., Chandra M., Ashby J., Chow A., O’Connor S.T., Li S., Chin A.R., Somlo G., Palomares M., Li Z., Tremblay J.R., Tsuyada A., Sun G., Reid M.A., Wu X., Swiderski P., Ren X., Shi Y., Kong M., Zhong W., Chen Y, & Wang S.E. (2015). Breast cancer-secreted miR-122 reprograms glucose metabolism in pre-metastatic niche to promote metastasis. Nature cell biology, 17(2), 183-194.
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4

Sciatic Nerve Protein Extraction and Western Blot

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Sciatic nerves were dissected from mice, after removal of the epineurium and perineurium, the nerves were frozen in liquid nitrogen and conserved at -80°C. Protein were extracted from whole sciatic nerves after homogenization by sonication in standard RIPA lysis buffer and the protein concentration was measured using a BCA protein assay kit (Pierce). 20 μg proteins were directly analyzed by western blot using standard procedures with 12% SDS–PAGE and transferred on PVDF membranes for immunoblotting. Primary antibodies: Rabbit anti-Pkm1 1/2500 (NBP2-14833, Novus Biologicals); Rabbit anti-Pkm2 1/2500 (SAB4200095, Sigma Aldrich); Mouse anti-α-β-Actin 1/10000 (C1.AC-15, #A1978, Sigma Aldrich). Secondary antibodies, Peroxidase Goat anti Rabbit or Mouse (H+L) (Jackson Immuno Research) were used at 1/10000 dilution.
Deck M., Van Hameren G., Campbell G., Bernard-Marissal N., Devaux J., Berthelot J., Lattard A., Médard J.J., Gautier B., Guelfi S., Abbou S., Quintana P., Chao de la Barca J.M., Reynier P., Lenaers G., Chrast R, & Tricaud N. (2022). Physiology of PNS axons relies on glycolytic metabolism in myelinating Schwann cells. PLoS ONE, 17(10), e0272097.
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