Anti wbp2

Manufactured by Proteintech

Sourced in United States

Anti-WBP2 is an antibody product that specifically targets the WBP2 protein. WBP2 is a transcriptional coactivator involved in the regulation of gene expression. This antibody can be used for the detection and study of the WBP2 protein in various biological samples and applications.

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Lab products found in correlation

Anti p akt, by Abcam (2 mentions) Ripa buffer, by Thermo Fisher Scientific (2 mentions) Nitrocellulose membrane, by Beyotime (1 mentions) Odyssey infrared scanning system, by LI COR (1 mentions) Ripa lysis buffer, by Beyotime (1 mentions) Bca protein assay kit, by Beyotime (1 mentions) Phenyl methyl sulfonyl fluoride (pmsf), by Beyotime (1 mentions) Anti akt, by Proteintech (1 mentions) Anti β actin, by ABclonal (1 mentions) Anti β actin, by Santa Cruz Biotechnology (1 mentions) Halt phosphatase inhibitor cocktail, by Thermo Fisher Scientific (1 mentions)

4 protocols using anti wbp2

Quantification of Protein Expression by Western Blot

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Total proteins were extracted by RIPA lysis buffer (Beyotime, China) together with PMSF (Beyotime, China). The concentration of total protein was determined by BCA protein assay kit (Beyotime, China). After being separated by 10% sodium dodecyl sulfate-polyacrylamide (SDS-PAGE) gels, proteins lysates were transferred to nitrocellulose membranes (Beyotime, China). The membranes were blocked with 5% non-fatty milk for 1 h at room temperature and then immunoblotted at 4 °C overnight with primary antibodies: anti-WBP2 (1:1000, Proteintech, USA), anti-AKT (1:1000, Proteintech, USA), anti-p-AKT (1:1000, Abcam, USA) and anti-β-actin (1:10,000, Abclonal, China). After incubated with diluted secondary antibodies for 1 h at room temperature, the bands were scanned and analyzed by Odyssey Infrared scanning system (LI-COR Biosciences, USA). Original western blots were shown in Fig. S1.

Wang X., Song H., Fang L, & Wu T. (2022). EIF4A3-mediated circPRKCI expression promotes triple-negative breast cancer progression by regulating WBP2 and PI3K/AKT signaling pathway. Cell Death Discovery, 8, 92.

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Quantifying WBP2 in Mouse Sperm

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The amount of WBP2 present in a single mouse spermatozoon was estimated based on the densitometric comparison of known concentrations of recombinant WBP2 and known amounts of mouse spermatozoa using anti-WBP2 (ProteinTech) immunoblotting. The immunoreactivity of the bands was analyzed using ImageJ software and a standard curve of recWBP2 plotted using Microsoft Excel. The curve was used to extrapolate the WBP2 protein concentration per single mouse spermatozoon.

Hamilton L.E., Suzuki J., Acteau G., Shi M., Xu W., Meinsohn M.C., Sutovsky P, & Oko R. (2018). WBP2 shares a common location in mouse spermatozoa with WBP2NL/PAWP and like its descendent is a candidate mouse oocyte-activating factor. Biology of Reproduction, 99(6), 1171-1183.

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Western Blot Analysis of WBP2 Protein

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Cell pellets were lysed using RIPA buffer (ThermoFisher, cat. 89900) supplemented with ETDA complete miniprotease inhibitor cocktail (Sigma, cat. 11836170001), and HALT phosphatase inhibitor cocktail (ThermoFisher, cat. 78426). Protein was quantified by BCA assay prior to gel loading. Samples were then boiled for 5-10 min with Laemelli 6× loading dye and 1,4-dithiothreitol, then loaded into 12 or 15-well 4-12% gradient gels (Novus biologicals). Gels were run in 1× running buffer (Novus Biologicals) at 120 volts for 2 h and transferred in 1× transfer buffer (Novus Biologicals) with 10% methanol at 45 volts for 75 min. Gels were blocked in Odyssey blocking buffer diluted 1:1 with PBS. Gels were incubated with anti-WBP2 (rabbit polyclonal; Proteintech cat. 12030-1-AP) and anti-β-actin (mouse monoclonal; Santa Cruz, cat. sc-47778) overnight in blocking buffer. Gels were washed with PBS-T, incubated with anti-mouse 680, and anti-rabbit 800 secondary antibodies, and imaged using a Li-Cor Imager. Images were quantified using ImageStudioLite.

Field N.S., Elbulok O.A., Dybas J.M., Moser E.K., Dar A.A., Spruce L.A., Fazelinia H., Seeholzer S.H, & Oliver P.M. (2020). Itch attenuates CD4 T-cell proliferation in mice by limiting WBP2 protein stability. European journal of immunology, 50(10), 1468-1483.

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Immunohistochemical Analysis of Tumor Tissue

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Tumor tissue collected from the BALB/c nude mice was fixed in 4% paraformaldehyde, dehydrated by ethanol solution, embedded in paraffin and sectioned into 4 μm slides. Then the slides were incubated with anti-WBP2 (Proteintech, USA) and anti-p-AKT (Abcam, USA). Images were photographed by Leica Microsystems (Germany).

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