Anti mbd2

Manufactured by Abcam

Sourced in United States

Anti-MBD2 is a primary antibody that recognizes the MBD2 (Methyl-CpG Binding Domain Protein 2) protein. MBD2 is a member of the methyl-CpG binding domain (MBD) protein family and plays a role in transcriptional regulation and epigenetic modification of DNA.

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Lab products found in correlation

Recombinant human tgf β1, by Proteintech (2 mentions) Anti collagen 1, by Abcam (2 mentions) Anti fibronectin, by Abcam (2 mentions) Anti α smooth muscle actin α sma, by Boster Bio (1 mentions) Alexa fluor 594 labeled anti mouse rabbit, by Thermo Fisher Scientific (1 mentions) Alexa fluor 488 conjugated anti rabbit mouse antibodies, by Thermo Fisher Scientific (1 mentions) Anti ki67 primary antibody, by Abcam (1 mentions) Pannoramic midi, by 3DHISTECH (1 mentions) Anti collagen 4, by Abcam (1 mentions) Anti p erk1 2, by Abcam (1 mentions) Anti smad3, by Proteintech (1 mentions) Anti glyceraldehyde 3 phosphate dehydrogenase anti gapdh, by Santa Cruz Biotechnology (1 mentions) Anti p smad3, by Proteintech (1 mentions) Anti egr1, by Proteintech (1 mentions) Anti tgf β, by Abcam (1 mentions) Anti erk1 2, by Abcam (1 mentions) Anti collagen 4, by Proteintech (1 mentions) Anti gapdh, by Santa Cruz Biotechnology (1 mentions) Anti f4 80, by Proteintech (1 mentions) Anti g0s2, by Proteintech (1 mentions)

4 protocols using anti mbd2

Immunofluorescence Assay for Lung Tissue Analysis

Cited 2 times

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The immunofluorescence assay was performed as previously described (13 (link), 29 (link)). Human lung sections, rats’ lung sections (13 (link)), and HPASMCs were first stained with anti-α-smooth muscle actin (αSMA, Boster, Wuhan), anti-MBD2 (Abcam), and anti-Ki67 primary antibodies (Abcam), followed by staining with Alexa Fluor 594-labeled anti-mouse/rabbit, or Alexa Fluor 488-conjugated anti-rabbit/mouse antibodies (Invitrogen, Carlsbad, CA, USA), respectively. DAPI was used for staining the cell nuclei. A Pannoramic MIDI (3Dhistech, Budapest, Hungary) was used to scan and analyze the images. Mean fluorescence intensity was assessed by ImageJ software (NIH, Bethesda, MD).

Wu J., Huang Q., Li Q., Gu Y., Zhan Y., Wang T., Chen J., Zeng Z., Lv Y., Zhao J., Xia J, & Xie J. (2022). Increased Methyl-CpG-Binding Domain Protein 2 Promotes Cigarette Smoke-Induced Pulmonary Hypertension. Frontiers in Oncology, 12, 879793.

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Antibody Sources and Plasmid Construction

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Antibodies were obtained from multiple sources: anti-glyceraldehyde 3-phosphate dehydrogenase (anti-GAPDH) and anti-AP1 from Santa Cruz Biotechnology (Santa Cruz, CA, USA); anti-MBD2, anti-collagen I, anti-collagen IV, anti-TGF-β, anti-ERK1/2, anti-p-ERK1/2, and anti-fibronectin from Abcam (Cambridge, MA, USA); and anti-EGR1, anti-SMAD3, and anti-p-SMAD3 from Proteintech Group (Rosemont, IL, USA),NCM Universal Antibody Diluent (New Cell and Molecular Biotech,Suzhou, China). The recombinant human TGF-β1 was obtained from Proteintech Group. The plasmids containing the methylation promoter of EGR1 CpG-free pCpGI luciferase reporter, MBD2 and mtMBD2 (the deletion of the methylated DNA binding domain), were constructed by the Ruqi Biology (Guangdong, Guangzhou, China), according to previously published reports.²⁸ (link)^,³⁹ (link)

Ai K., Li X., Zhang P., Pan J., Li H., He Z., Zhang H., Yi L., Kang Y., Wang Y., Chen J., Li Y., Xiang X., Chai X, & Zhang D. (2022). Genetic or siRNA inhibition of MBD2 attenuates the UUO- and I/R-induced renal fibrosis via downregulation of EGR1. Molecular Therapy. Nucleic Acids, 28, 77-86.

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Antibody and Plasmid Sources for Cell Studies

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Antibodies were obtained from multiple sources: anti-GAPDH from Santa Cruz Biotechnology (Santa Cruz, CA, USA), anti-MBD2, anti-collagen I, anti-α-SMA and anti-fibronectin from Abcam (Cambridge, MA, USA), and anti-F4/80, anti-collagen IV, and anti-G0S2 from Proteintech Group (Rosemont, IL, USA). The recombinant human TGF-β1 was obtained from Proteintech Group. The plasmids containing the methylation promoter of the G0S2 CpG-free pCpGI luciferase reporter, MBD2, and mtMBD2 (the deletion of the methylated DNA binding domain), were constructed by the Ruqi Biology Company (Guangdong, Guangzhou, China) according to previously published reports [15 (link), 17 (link)]. The MBD2 siRNA was designed and synthesized by the Ruibo Biology Company (Guangdong, Guangzhou, China) as described in our previously published paper [18 (link)].

Ai K., Pan J., Zhang P., Li H., He Z., Zhang H., Li X., Li Y., Yi L., Kang Y., Wang Y., Xiang X., Chai X, & Zhang D. (2022). Methyl-CpG-binding domain protein 2 contributes to renal fibrosis through promoting polarized M1 macrophages. Cell Death & Disease, 13(2), 125.

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ChIP and MNase Assays for Epigenetic Profiling

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ChIP and MNase assays were performed as described55 56 (link). Briefly, 100 mg of eAT were cross-linked with 1% formaldehyde for 15 min at 37 °C. For ChIP assay, sonicated chromatin was immuno-precipitated with the following antibodies: anti-p300 (#SC-585, Santa Cruz Biotechnology), anti-Ac-H4-K16 (#07-329, Millipore, Temecula, CA), anti-DNMT1 (#NB100-56519) and anti-DNMT3b (#NB300-516) from Novus Biologicals (Littleton, CO), anti-DNMT3a (#ab2850), anti-MBD2 (#ab38646), and anti-RNA Pol II (#ab5408) from Abcam and anti-rabbit IgG (#I8140) and anti-mouse IgG (#I8765) from Sigma-Aldrich. For MNase assay, nuclei were isolated from 100 mg of eAT, suspended in wash buffer (100 mmol/L Tris-HCl, 15 mmol/L NaCl, 60 mmol/L KCl, 1 mmol/L CaCl₂) and treated with 200 U of MNase for 20 min at 37 °C. Cross-link reversal was performed at 65 °C for at least 16 h followed by an RNase and subsequent proteinase K digestion. DNA was purified by phenol–chloroform. Samples were then run on 1% agarose gel and the resulting mononucleosomal DNA fragments (~150 bp) were gel purified. For both assays, relative protein binding and nucleosome occupancy to the Ankrd26 gene were evaluated on recovered DNA by qPCR. Samples were normalized to their respective input using the 2^−ΔCT method.

Raciti G.A., Spinelli R., Desiderio A., Longo M., Parrillo L., Nigro C., D’Esposito V., Mirra P., Fiory F., Pilone V., Forestieri P., Formisano P., Pastan I., Miele C, & Beguinot F. (2017). Specific CpG hyper-methylation leads to Ankrd26 gene down-regulation in white adipose tissue of a mouse model of diet-induced obesity. Scientific Reports, 7, 43526.

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