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Western blotAntibody data
- Antibody Data
- Antigen structure
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- Validations
- Western blot [5]
- Immunocytochemistry [2]
- Immunohistochemistry [1]
- Chromatin Immunoprecipitation [1]
- Other assay [1]
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- Product number
- PA5-29106 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- MCM3 Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Recombinant protein fragment
- Description
- Recommended positive controls: 293T, A431, H1299, HeLa, HepG2, Molt-4, Raji.
- Concentration
- 0.71 mg/mL
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western Blot using MCM3 Polyclonal Antibody (Product # PA5-29106). Sample (30 µg of whole cell lysate). Lane A: Hep G2 . 7.5% SDS PAGE. MCM3 Polyclonal Antibody (Product # PA5-29106) diluted at 1:1,000.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western Blot using MCM3 Polyclonal Antibody (Product # PA5-29106). Various whole cell extracts (30 µg) were separated by 7.5% SDS-PAGE, and the membrane was blotted with MCM3 Polyclonal Antibody (Product # PA5-29106) diluted at 1:500. 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 MCM3 Polyclonal Antibody (Product # PA5-29106). Mouse tissue extract (50 µg) was separated by 7.5% SDS-PAGE, and the membrane was blotted with MCM3 Polyclonal Antibody (Product # PA5-29106) diluted at 1:500. 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 MCM3 was achieved by transfecting HeLa with MCM3 specific siRNAs (Silencer® select Product # s8589, s8590). Western blot analysis (Fig. a) was performed using modified whole cell extracts (1% SDS) from the MCM4 knockdown cells (Lane 3), non-specific scrambled siRNA transfected cells (Lane 2) and untransfected cells (Lane 1). The blot was probed with MCM3 Polyclonal Antibody (Product # PA5-29106, 1:1000 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 i histogram (Fig. b). Loss of signal upon siRNA mediated knock down confirms that antibody is specific to MCM3.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using MCM3 Polyclonal Antibody (Product # PA5-29106) and a 90 kDa band corresponding to MCM3 was observed across cell lines tested along with an uncharacterized band at around 70 kDa. Whole cell extracts (30 µg lysate) of BJ (Lane 1), HeLa (Lane 2),A549 (Lane 3), MCF-7 (Lane 4) and Hep G2 (Lane 5) were electrophoresed using 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 (1:1000 dilution) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000 dilution) (Product # A27014) 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 MCM3 was performed in HeLa cells fixed in 4% paraformaldehyde at RT for 15 min. Green: MCM3 Polyclonal Antibody (Product # PA5-29106) diluted at 1:500. Red: alpha Tubulin, a cytoskeleton marker. Blue: Hoechst 33342 staining.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunocytochemistry-Immunofluorescence analysis of MCM3 was performed in HeLa cells fixed in 4% paraformaldehyde at RT for 15 min. Green: MCM3 Polyclonal Antibody (Product # PA5-29106) diluted at 1:500. Red: alpha Tubulin, a cytoskeleton marker. Blue: Hoechst 33342 staining.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- MCM3 Polyclonal Antibody detects MCM3 protein at nucleus on mouse colon by immunohistochemical analysis. Sample: Paraffin-embedded mouse colon. MCM3 Polyclonal Antibody (Product # PA5-29106) diluted at 1:1,000. Antigen Retrieval: EDTA based buffer, pH 8.0, 15 min.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Cross-linked ChIP was performed with HeLa chromatin extract and 5 µg of either control rabbit IgG or MCM3 Polyclonal Antibody (Product # PA5-29106). The precipitated DNA was detected by PCR with primer set targeting to Myc promoter.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- MCM3 Polyclonal Antibody immunoprecipitates MCM3 protein in IP experiments. IP samples: HepG2 whole cell extract. A. 40 µg HepG2 whole cell extract. B. Control with 4 µg of preimmune Rabbit IgG. C. Immunoprecipitation of MCM3 protein by 4 µg MCM3 Polyclonal Antibody (Product # PA5-29106). 7.5 % SDS-PAGE. The immunoprecipitated MCM3 protein was detected by MCM3 Polyclonal Antibody (Product # PA5-29106) diluted at 1:1,000.
