Anti menin

Manufactured by Fortis Life Sciences

Sourced in United States

Anti-Menin is a laboratory equipment product designed for research purposes. It functions as a protein that binds to the Menin protein, which is involved in various cellular processes. The core function of Anti-Menin is to facilitate the study and investigation of Menin-related mechanisms and pathways. Further details on its intended use are not provided.

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Lab products found in correlation

Anti gapdh, by Cell Signaling Technology (2 mentions) Anti nf ya, by Santa Cruz Biotechnology (2 mentions) Anti h3k4me1, by Abcam (2 mentions) Anti actin, by Abcam (2 mentions) Hrp conjugated anti mouse or anti rabbit secondary antibodies, by GE Healthcare (2 mentions) Anti h3, by Abcam (2 mentions) Tween 20, by Merck Group (2 mentions) Pvdf membrane, by Merck Group (2 mentions) Anti hsp90, by BD (2 mentions) Anti ha, by BioLegend (2 mentions) Sybr premix ex taq 2, by Takara Bio (1 mentions) Anti β actin, by Santa Cruz Biotechnology (1 mentions) Tris acetate gel, by Thermo Fisher Scientific (1 mentions) Anti dot1l, by Cell Signaling Technology (1 mentions) Semi dry transfer, by Thermo Fisher Scientific (1 mentions) Anti h3k79me2, by Abcam (1 mentions) Western blot detection kit, by AbFrontier (1 mentions) Anti ar, by Santa Cruz Biotechnology (1 mentions) Anti α tubulin, by AbFrontier (1 mentions) Anti lamin b1, by Thermo Fisher Scientific (1 mentions)

8 protocols using anti menin

Menin ChIP-qPCR of Cyclin D1

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Briefly, 5 × 10⁶ normal MAC-T cells were subjected to cross-linking with 1% formaldehyde, after which the cells were lysed in 1 ml of SDS lysis buffer and sonicated with a Bioruptor sonicator (UCD-200TM-EX, Diagenode, Belgium) to shear the chromatin into 200 – 1,000 bp fragments. The sonicated cell lysates (100 μl of the 1 ml supernatant) and 2 μg of antibodies were used for each immunoprecipitation. Anti-menin (Bethyl Laboratories, Montgomery, TX, USA) and anti-rabbit IgG (Sangon Biotech, Shanghai, China) were used for ChIP analyses. The precipitated DNA was used as template for quantitative real-time PCR using the SYBR-Premix Ex Taq II (TaKaRa, Dalian, China). Primer pairs screening of 1200 bp promoter region of Cyclin D1 gene were designed and synthesized, their detailed information were listed in Supplementary Table S3. For quantitative real-time PCR, CT values for each ChIP were obtained from triplicate reactions. The results were representative of three independent ChIP experiments, showing as the percentage of input by quantifying the amount of chromatin obtained from immunoprecipitation relative to the amount in the input samples.

Shi K., Liu X., Li H., Lin X., Yan Z., Cao Q., Zhao M., Xu Z, & Wang Z. (2017). Menin Modulates Mammary Epithelial Cell Numbers in Bovine Mammary Glands Through Cyclin D1. Journal of Mammary Gland Biology and Neoplasia, 22(4), 221-233.

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Antibody-based Protein Analysis Techniques

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Antibodies used were antimenin (Bethyl, Montgomery, Tex; A300-105A), anti-FoxO1 (CST, Boston, Mass; no. 2880), anti–p-AKT (CST; no. 4060), anti-AKT (CST; no. 9272), anti–p-FoxO1 (CST; no. 9461), anti-hemagglutinin (HA) (CWbiotech, Beijing, China; CW0092A), antiubiquitin (CST; no. 3936), anti-Skp2 (CST; no. 2652), anti–caspase-3 (CST; no. 9665), and anti–β-actin (Santa Cruz Biotechnology, Dallas, Tex; SC47778).

Jiang Z., Xing B., Feng Z., Ma J., Ma X, & Hua X. (2019). Menin Upregulates FOXO1 Protein Stability by Repressing Skp2-Mediated Degradation in β Cells. Pancreas, 48(2), 267-274.

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Western Blot Analysis of Epigenetic Regulators

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5×10⁴ PBS washed cells were lysed in 50 μL of 1× loading buffer supplemented with 10 mM DTT and denatured for 10 min at 95°C. Samples were loaded (20 μL per lane) onto either 10% Bis-Tris or Tris-Acetate gels (ThermoFisher), proteins were separated by electrophoresis for 2 hr then transferred onto nitrocellulose membrane using semi-dry transfer (ThermoFisher). The membranes were blocked in 5% dry milk for 30 min and incubated overnight with anti-Menin (Bethyl), anti-DOT1L (Cell Signaling), anti-H3K79me2 (Abcam) or anti-GAPDH (Cell Signaling) antibody. The next day membranes were washed in TBST and developed using a secondary rabbit anti-HRP (Cell Signaling) and chemiluminescence kit (Pierce).

Krivtsov A.V., Evans K., Gadrey J.Y., Eschle B.K., Hatton C., Uckelmann H.J., Ross K.N., Perner F., Olsen S.N., Pritchard T., McDermott L., Jones C.D., Jing D., Braytee A., Chacon D., Earley E., McKeever B.M., Claremon D., Gifford A.J., Lee H.J., Teicher B.A., Pimanda J.E., Beck D., Perry J.A., Smith M.A., McGeehan G.M., Lock R.B, & Armstrong S.A. (2019). A Menin-MLL inhibitor induces specific chromatin changes and eradicates disease in models of MLL-rearranged leukemia. Cancer cell, 36(6), 660-673.e11.

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Western Blot Analysis of Epigenetic Factors

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Whole cell lysates were separated by SDS-PAGE, transferred to a PVDF membrane (EMD Millipore), blocked in 5% non-fat milk in TBS plus 0.5% Tween-20 (Sigma-Aldrich), probed with primary antibodies, and detected with HRP-conjugated anti-rabbit or anti-mouse secondary antibodies (GE Healthcare). Primary antibodies used included: anti-Cas9 (CST, 14697), anti-UTX (CST, 33510), anti-Menin (Bethyl, A300-105A), anti-NF-YA (Santa Cruz Biotechnology, sc-17753), anti-Actin (abcam, ab8224), anti-HSP90 (BD Biosciences, 610418), anti-HA (Biolegend, 901501), anti-H3K4me1 (abcam, ab8895), anti-H3K4me3 (CST, 9751), anti-H3 (abcam, ab1791).

Soto-Feliciano Y.M., Sánchez-Rivera F.J., Perner F., Barrows D.W., Kastenhuber E.R., Ho Y.J., Carroll T., Xiong Y., Anand D., Soshnev A.A., Gates L., Beytagh M.C., Cheon D., Gu S., Liu X.S., Krivtsov A.V., Meneses M., de Stanchina E., Stone R.M., Armstrong S.A., Lowe S.W, & Allis C.D. (2022). A Molecular Switch between Mammalian MLL Complexes Dictates Response to Menin–MLL Inhibition. Cancer Discovery, 13(1), 146-169.

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Western Blot Analysis of Cellular Proteins

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Cells were harvested, washed twice with cold PBS, and lysed using a lysis buffer consisting of 25 mM Tris HCl (pH 8.0), 1 mM EDTA, 150 mM NaCl, and 0.5% NP40. After a 10-min incubation on ice, the cell lysate was centrifuged at 13,000 rpm at 4°C for 10 min. Equal amounts of protein from extracts were separated using SDS-PAGE and transferred to nitrocellulose membranes. The blots were blocked with 5% skim milk and incubated with primary antibodies, which were diluted in the range of 1:1,000 to 1:5,000. After washing with TBS-T (5 mM Tris HCl [pH 7.4], 150 mM NaCl, and 0.01% Tween 20), secondary antibodies were added at a dilution of 1:10,000 for a 1-h incubation at room temperature. Protein signals were detected using a western blot detection kit (AbFrontier, Korea). The following primary antibodies were used in this study: anti-β-actin (Merck Millipore, USA), anti-α-tubulin (AbFrontier), anti-menin (Bethyl Laboratories, USA), anti-cleaved-PARP (Cell Signaling Technology, USA), anti-AR (Santa Cruz Biotechnology, USA), and anti-Lamin B1 (Thermo Fisher Scientific).

Kim T., Jeong K., Kim E., Yoon K., Choi J., Park J.H., Kim J.H., Kim H.S., Youn H.D, & Cho E.J. (2022). Menin Enhances Androgen Receptor-Independent Proliferation and Migration of Prostate Cancer Cells. Molecules and Cells, 45(4), 202-215.

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Histone Modification Antibody Panel

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The following antibodies are used in this study: anti-H3K4me1 (Cell Signaling Technology [CST], 5326), anti-H3K4me2 (generated in house), anti-H3K4me3 (CST, 9751), anti-H3K27ac (CST, 8173), H3K27me3 (CST, 9733), anti-MLL3 NT (generated in house), anti-MLL3 MT (generated in house), anti-MLL4 NT (generated in house), anti-MLL4 CT (generated in house), anti-MLL2 (generated in house), anti-SET1A (generated in house), anti-Menin (CST, 6891), anti-MLL1C (CST, 14197), anti-RBBP5 (Bethyl Laboratories, A300-109A), anti-NCOA6 (Bethyl Laboratories, A300-410A), anti-UTX (CST, 33510), anti-PTIP (Bethyl Laboratories, A300-370A), ASH2L (CST, 5019), H3 Ser10-p (CST 53348), CDT1 (CST, 8064), Cyclin B1 (CST, 12231), Geminin (CST, 52508), Cyclin E1 (CST, 20808), Cyclin A2 (CST, 91500), p-cdc2 (CST, 4539), PARP (CST, 9542) Caspase3 (CST, 9662), anti-GART (Santa Cruz Biotechnology, sc-166447), anti-PAICS (Proteintech, 12967-1-AP), Hsp90 (Santa Cruz Biotechnology, sc-13119), and anti-β-tubulin (Developmental Studies Hybridoma Bank, E7). Western blot analysis was performed as previously described (53 (link)).

Zhao Z., Cao K., Watanabe J., Philips C.N., Zeidner J.M., Ishi Y., Wang Q., Gold S.R., Junkins K., Bartom E.T., Yue F., Chandel N.S., Hashizume R., Ben-Sahra I, & Shilatifard A. (2023). Therapeutic targeting of metabolic vulnerabilities in cancers with MLL3/4-COMPASS epigenetic regulator mutations. The Journal of Clinical Investigation, 133(13), e169993.

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Western Blot Analysis of Metabolic Enzymes

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Western blot was performed according to established protocols with the following primary antibodies: anti-PHGDH 1:1000 (3PGDH 6B2; Santa Cruz Biotechnology, sc100–317); anti-PSAT1 1:1000 (Thermo Fisher PA5–22124); anti-GAPDH 1:1000 (Cell Signaling 14C10); monoclonal anti-GAPDH 1:1000 (Thermo Fisher AM4300); anti-menin 1:1000 (Bethyl A300–105A). Membranes were probed with the appropriate secondary antibodies from LiCor Biotechnology (Lincoln, NE) at 1:10,000 dilution and imaged with a LiCor Odyssey infrared imager.

Svoboda L.K., Teh S.S., Sud S., Kerk S., Zebolsky A., Treichel S., Thomas D., Halbrook C.J., Lee H.J., Kremer D., Zhang L., Klossowski S., Bankhead A.R., Magnuson B., Ljungman M., Cierpicki T., Grembecka J., Lyssiotis C.A, & Lawlor E.R. (2018). Menin regulates the serine biosynthetic pathway in Ewing sarcoma. The Journal of pathology, 245(3), 324-336.

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Western Blot Analysis of Chromatin Regulators

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Whole-cell lysates were separated by SDS-PAGE, transferred to a PVDF membrane (EMD Millipore), blocked in 5% nonfat milk in TBS plus 0.5% Tween-20 (Sigma-Aldrich), probed with primary antibodies, and detected with HRP-conjugated anti-rabbit or anti-mouse secondary antibodies (GE Healthcare). Primary antibodies used included anti-Cas9 (Cell Signaling Technology, 14697), anti-UTX (Cell Signaling Technology, 33510), anti-Menin (Bethyl, A300-105A), anti-NF-YA (Santa Cruz Biotechnology, sc-17753), anti-Actin (Abcam, ab8224), anti-HSP90 (BD Biosciences, 610418), anti-HA (BioLegend, 901501), anti-H3K4me1 (Abcam, ab8895), anti-H3K4me3 (Cell Signaling Technology, 9751), and anti-H3 (Abcam, ab1791).

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