Anti meis1

Manufactured by Abcam

Anti-MEIS1 # is a primary antibody that specifically binds to the MEIS1 protein. MEIS1 is a transcription factor involved in the regulation of various cellular processes. This antibody can be used for the detection and analysis of MEIS1 in a range of applications, such as Western blotting and immunohistochemistry.

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Lab products found in correlation

Lipofectamine 2000, by Thermo Fisher Scientific (2 mentions) Pierce bca protein assay kit, by Thermo Fisher Scientific (2 mentions) Puromycin, by Thermo Fisher Scientific (1 mentions) Virapower lentiviral packaging mix, by Thermo Fisher Scientific (1 mentions) Nitrocellulose, by LI COR (1 mentions) Sample buffer, by Bio-Rad (1 mentions) Donkey anti rabbit irdye 680, by LI COR (1 mentions) Odyssey infrared scanner, by LI COR (1 mentions) Irdye 800cw goat anti mouse, by LI COR (1 mentions) Anti β actin, by Merck Group (1 mentions) β mercaptoethanol, by Merck Group (1 mentions) Polyvinylidene difluoride membrane, by Merck Group (1 mentions) Pe conjugated anti annexin 5 and 7 aad, by BD (1 mentions)

3 protocols using anti meis1

Lentiviral Overexpression of MEIS1 and MEIS2

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To derive cell lines overexpressing lentiviral constructs of MEIS1 or MEIS2, all lentiviral vectors used in this study contain the gene-of-interest in the pReceiver-LV105 backbone (GeneCopoeia). High titer lentivirus was made by separately co-transfecting the LV105 constructs with ViraPower Lentiviral packaging mix (#K497500, Thermo Fisher Scientific) in HEK-293T cells using Lipofectamine 2000 (#11668019, Thermo Fisher Scientific) according to manufacturer's instructions. After 48 and 72 hr, media containing the lentivirus was collected, spun down, and filtered using a 0.45 mm filter and used to infect target CWR-22rv1 or LAPC-4 cells with 5 mg/mL polybrene for 48 hr. Complete media were then replaced followed by selection and maintenance with puromycin (1 mg/mL, Invitrogen). Confirmation of MEIS1 or MEIS2 expression was confirmed using both qRT-PCR and Western blotting (anti-MEIS1 #ab19867, abcam); anti-MEIS2 (#TA337288, OriGene).

VanOpstall C., Perike S., Brechka H., Gillard M., Lamperis S., Zhu B., Brown R., Bhanvadia R, & Vander Griend D.J. (2020). MEIS-mediated suppression of human prostate cancer growth and metastasis through HOXB13-dependent regulation of proteoglycans. eLife, 9, e53600.

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Western Blot Analysis of MEIS Proteins

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Whole-cell lysates of 100,000 or more cells were used per lane. Western blotting was performed as previously reported (15 (link)). Briefly, cells were rinsed with cold PBS and scraped into RIPA buffer supplemented with protease inhibitors, sonicated twice and re-suspended in 4× sample buffer (BioRad; Hercules, CA) supplemented with 10% Beta-mercaptoethanol (Sigma-Aldrich; St. Louis MO). The Pierce BCA protein assay kit (Thermo Fisher Scientific; Grand Island, NY) was used to determine protein concentration. 60 ug of protein was loaded on a 10% SDS-polyacrylamide gel and electrophoresed, and transferred to nitrocellulose (LI COR, Odyssey; Lincoln, NE) overnight at 4°C. The membrane was blocked overnight in 5% non-fat milk in TBS at 4°C. Primary antibodies used were: anti-MEIS1 (Ab19867, Abcam; Cambridge, MA), anti-MEIS2 (ARP34683_P050, Aviva Systems Biology; San Diego, CA), anti-Beta Actin, (Clone AC-15, Sigma-Aldrich; St. Louis, MO). The secondary antibodies goat anti-mouse IRDye 800 CW or donkey anti-rabbit IRDye 680 from LI-COR Biosciences were used, and images captured using an infrared Odyssey scanner (LI-COR).

Bhanvadia R.R., VanOpstall C., Brechka H., Barashi N.S., Gillard M., McAuley E.M., Vasquez J.M., Paner G., Chan W.C., Andrade J., De Marzo A.M., Han M., Szmulewitz R.Z, & Vander Griend D.J. (2018). MEIS1 and MEIS2 Expression and Prostate Cancer Progression: A Role For HOXB13 Binding Partners in Metastatic Disease. Clinical cancer research : an official journal of the American Association for Cancer Research, 24(15), 3668-3680.

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Protein Expression and Apoptosis Analysis

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Plasmids of pCMV-SPORT6-Meis1, Meis1∆PIM, pCMV-SPORT6-HoxA9 and Hoxa∆MID were transfected into 293 T cells. Whole cell lysates were electrophoresed on 8–10 % sodium dodecyl sulfate polyacrylamide gels and transferred onto polyvinylidene difluoride membranes (Millipore). The membranes were incubated with primary antibodies overnight at 4 °C followed by incubation with appropriate horseradish peroxidase-conjugated secondary antibodies. The following antibodies were used: anti-Meis1 (Abcam), anti-HoxA9 (Proteintech) and anti-β-actin (Calbiochem). For analysis of apoptosis, cells were stained with PE conjugated anti-annexin V and 7-AAD (BD Pharmingen) according to the manufacturer’s instructions.

Kocabas F., Xie L., Xie J., Yu Z., DeBerardinis R.J., Kimura W., Thet S., Elshamy A.F., Abouellail H., Muralidhar S., Liu X., Chen C., Sadek H.A., Zhang C.C, & Zheng J. (2015). Hypoxic metabolism in human hematopoietic stem cells. Cell & Bioscience, 5, 39.

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