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Western blotAntibody data
- Antibody Data
- Antigen structure
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- Validations
- Western blot [9]
- Immunocytochemistry [1]
- Immunohistochemistry [1]
- Other assay [2]
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- Product number
- PA5-77945 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- DNMT3A Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Recombinant full-length protein
- Description
- Positive Control: 293T, HeLa, HepG2, Neuro2A, PC-12, Rat2, CL1-0
- Concentration
- 1.28 mg/mL
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of DNMT3A in whole cell lysate using 30 µg of protein. Samples were separated with 5% SDS-PAGE and incubated with DNMT3A polyclonal antibody (Product # PA5-77945) using a dilution of 1:2000 followed by HRP-conjugated anti-rabbit IgG.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of DNMT3A in non-transfected (-) and transfected (+) 293T cells using 30 µg of protein. Samples were separated with 5% SDS-PAGE and incubated with DNMT3A polyclonal antibody (Product # PA5-77945) using a dilution of 1:500 followed by HRP-conjugated anti-rabbit IgG.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of DNMT3A in whole cell lysate using 30 µg of protein. Samples were separated with 7.5% SDS-PAGE and incubated with DNMT3A polyclonal antibody (Product # PA5-77945) using a dilution of 1:1000 followed by HRP-conjugated anti-rabbit IgG.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of DNMT3A in whole cell lysate using 30 µg of protein. Samples were separated with 7.5% SDS-PAGE and incubated with DNMT3A polyclonal antibody (Product # PA5-77945) using a dilution of 1:1000 followed by HRP-conjugated anti-rabbit IgG.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western Blot using DNMT3A Polyclonal Antibody (Product # PA5-77945). Various whole cell extracts (30 µg) were separated by 7.5% SDS-PAGE, and the membrane was blotted with DNMT3A Polyclonal Antibody (Product # PA5-77945) diluted at 1:1,000. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western Blot using DNMT3A Polyclonal Antibody (Product # PA5-77945). Whole cell extract (30 µg) was separated by 7.5% SDS-PAGE, and the membrane was blotted with DNMT3A Polyclonal Antibody (Product # PA5-77945) diluted at 1:1,000. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western Blot using DNMT3A Polyclonal Antibody (Product # PA5-77945). Various whole cell extracts (30 µg) were separated by 5% SDS-PAGE, and the membrane was blotted with DNMT3A Polyclonal Antibody (Product # PA5-77945) diluted at 1:2,000. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Knockdown of DNMT3A was achieved by transfecting NTERA-2 cl.D1 with DNMT3A specific siRNAs (Silencer® select Product # s200426, s502236). Western blot analysis (Fig. a) was performed using modified whole cell extracts (1% SDS) from the DNMT3A knockdown cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with DNMT3A Polyclonal Antibody (Product # PA5-77945, 1:2000 dilution) and Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 0.25 µg/mL, 1:4000 dilution). Densitometric analysis of this western blot is shown in histogram (Fig. b). Decrease in signal upon siRNA mediated knock down confirms that antibody is specific to DNMT3A.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Anti-DNMT3A Polyclonal Antibody, (Product # PA5-77945) and a 110 kDa band corresponding to DNMT3A was observed in the cell lines, with higher expression in NTERA-2 compared to HeLa and Jurkat. Modified whole cell extracts (1% SDS) (40 µg lysate) of NTERA-2 (Lane 1), HeLa (Lane 2), MCF7 (Lane 3), Jurkat (Lane 4), NCCIT (lane 5) and iPSC (Lane 6) were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris gel (Product # NP0321BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:2000 dilution) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L), Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunocytochemistry-Immunofluorescence analysis of DNMT3A was performed in 293T cells fixed in 4% paraformaldehyde at RT for 15 min. Green: DNMT3A Polyclonal Antibody (Product # PA5-77945) diluted at 1:1000. Red: phalloidin, a cytoskeleton marker. Scale bar = 10 µm.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- DNMT3A Polyclonal Antibody detects DNMT3A protein at nucleus on mouse ovary by immunohistochemical analysis. Sample: Paraffin-embedded mouse ovary. DNMT3A Polyclonal Antibody (Product # PA5-77945) dilution: 1:500. Antigen Retrieval: EDTA based buffer, pH 8.0, 15 min.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- DNMT3A Polyclonal Antibody immunoprecipitates DNMT3A protein in IP experiments. IP samples: 293T nuclear extract. A. 50 µg 293T nuclear extract. B. Control with 4 µg of preimmune Rabbit IgG. C. Immunoprecipitation of DNMT3A protein by 4 µg DNMT3A Polyclonal Antibody (Product # PA5-77945). 5 % SDS-PAGE. The immunoprecipitated DNMT3A protein was detected by DNMT3A Polyclonal Antibody (Product # PA5-77945) diluted at 1:2,000.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunoprecipitation of DNMT3A was performed in 293T whole cell extracts using 5 µg of DNMT3A Polyclonal Antibody (Product # PA5-77945). Samples were transferred to a membrane and probed with DNMT3A Polyclonal Antibody as a primary antibody and an HRP-conjugated anti-Rabbit IgG was used as a secondary antibody.
