- MA5-16171 - Invitrogen Antibodies DNMT3A antibody

Wang X, Zhang J, Liao Y, Jin Y, Yu X, Li H, Yang Q, Li X, Chen R, Wu D, Zhu H
Computational and mathematical methods in medicine 2022;2022:9212116

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis ofDnmt3a in (A) Dnmt3a transfected293 cell lysate and (B) untransfected 293 cell lysate using a Dnmt3a monoclonal antibody (Product # MA5-16171) at1 µg/mL.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis of DNMT3A in NTERA-2 cells. Samples were incubated in DNMT3A monoclonal antibody (Product # MA5-16171) using a dilution of 2 µg/mL followed by an anti-mouse HRP secondary antibody. Treated with Zebularine as indicated for 24 hours. Cell lysates were prepared and separated on a 7.5% gel by SDS-PAGE. Protein was transferred to PVDF membrane and blocked in 5% non-fat milk. Detection: chemiluminescence. Note the decrease in Dnmt3a expression upon treatment with 500 µM Zebularine (arrowhead). Additional bands at 90 and 75 kDa can also be detected with this antibody and may represent alternative splice variants. Tubulin is shown as a loading control.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of DNMT3A in (A) Dnmt3a transfected 293 cell lysate and (B) untransfected 293 cell lysate. Samples were incubated in DNMT3A monoclonal antibody (Product # MA5-16171 using a dilution of 1 µg/mL.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis was performed on modified whole cell extracts (1% SDS) (30 µg lysate) of HeLa (Lane 1), Jurkat (Lane 2) MCF7 (Lane 3), NCCIT (Lane 4), NTERA-2 (Lane 5), 3T3-L1 (Lane 6), HEK-293 (Lane 7), A549 (Lane 8), iPSC (Lane 9) and NIH/3T3 (Lane 10). The blot was probed with Anti- DNMT3A Monoclonal Antibody (Product # MA5-16171, 2 µg/mL) and detected by chemiluminescence using Goat anti Mouse IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A28177, 0.25 µg/mL, 1:4000 dilution). A 83 kDa band corresponding to DNMT3A was detected in NCCIT, NTERA-2 and iPSC when compared to other cell lines which are reported to be negative for DNMT3A expression.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunohistochemical analysis of DNMT3A in human hepatocellular carcinoma tissue section. Samples were incubated in DNMT3A monoclonal antibody (Product # MA5-16171) using a dilution of 1:100. The antibody generated a specific cytoplasmic staining in all the cancer cells while some of the cells depicted nuclear staining also.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Flow cytometry of DNMT3A in NTERA-2 cells (blue) and a matched isotype control (orange). Samples were incubated in DNMT3A monoclonal antibody (Product # MA5-16171) using a dilution of 1 µg/mL for 30 minutes at room temperature. Cells were fixed with 4% PFA and then permeablized with 0.1% saponin. Both antibodies were conjugated to Phycoerythrin.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Flow cytometry of DNMT3A in HepG2 cells. Samples were incubated in DNMT3A monoclonal antibody (Product # MA5-16171) using a dilution of 1 µg/mL for 30 minutes at room temperature. Antibody (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeablized with 0.1% saponin. Both antibodies were conjugated to allophycocyanin (APC).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Flow cytometry of DNMT3A in HeLa cells. Samples were incubated in DNMT3A monoclonal antibody (Product # MA5-16171) using a dilution of 1 µg/mL for 30 minutes at room temperature. Antibody (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Both antibodies were conjugated to allophycocyanin (APC).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Flow cytometry of DNMT3A in Jurkat cells. Samples were incubated in DNMT3A monoclonal (Product # MA5-16171) using a dilution of 2.5 µg/mL for 30 minutes at room temperature. Antibody (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeablized with 0.1% saponin. Both antibodies were conjugated to Alexa Fluor 647.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Flow cytometry of DNMT3A in 1 x 10^6 CHO (A) and HEK-293 (B) cells. Samples were incubated in DNMT3A monoclonal antibody (Product # MA5-16171) using a dilution of 1 µg/1x10^6 cells. Antibody (dark blue). Isotype control shown in orange.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 3 DNMT1-mediated DNA methylation was significantly increased in the early phase of diabetic retinopathy. (a) STZ injection upregulated the global DNA methylation level of the retina until 6 weeks after injection. (b) Quantification of the average 5-mC content in DNA methylation of the retina isolated from sodium citrate buffer injection (control) and STZ injection (STZ) groups. (c-d) Western blot showed the protein expression levels of DNMT1, DNMT3A, and DNMT3B. * P < 0.05, ** P < 0.01.

MA5-16171