Antibody data
- Antibody Data
- Antigen structure
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- Validations
- Western blot [4]
- Immunocytochemistry [2]
- Immunohistochemistry [2]
- Chromatin Immunoprecipitation [2]
- Other assay [1]
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Validation data
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- Product number
- PA5-35922 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- MCM6 Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Recombinant protein fragment
- Description
- Recommended positive controls: 293T, A431, HeLa, HepG2.
- Concentration
- 1 mg/mL
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
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- Experimental details
- MCM6 Polyclonal Antibody detects MCM6 protein by western blot analysis. A. 30 µg 293T whole cell lysate/extract. B. 30 µg A431whole cell lysate/extract. C. 30 µg HeLa whole cell lysate/extract. D. 30 µg HepG2 whole cell lysate/extract.7.5 % SDS-PAGE. MCM6 Polyclonal Antibody (Product # PA5-35922) dilution: 1:1,000.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- MCM6 Polyclonal Antibody detects MCM6 protein by western blot analysis. A. 30 µg 293T whole cell lysate/extract. B. 30 µg A431whole cell lysate/extract. C. 30 µg HeLa whole cell lysate/extract. D. 30 µg HepG2 whole cell lysate/extract.7.5 % SDS-PAGE. MCM6 Polyclonal Antibody (Product # PA5-35922) dilution: 1:1,000.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- KD of MCM6 was achieved by transfecting HeLa with MCM6 specific siRNAs (Silencer® select Product # s8599, s8598). Western blot analysis (Fig. a) was performed using whole cell extracts from the MCM6 KD cells (Lane 3), non-specific scrambled siRNA transfected cells (Lane 2) and untransfected cells (Lane 1). The blot was probed with MCM6 Polyclonal Antibody (Product # PA5-35922, 1µg/mL) and Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25µg/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 MCM6..
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- Invitrogen Antibodies (provider)
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- Experimental details
- Western blot was performed using Anti-MCM6 Polyclonal Antibody (Product # PA5-35922) and a 93 kDa band corresponding to MCM6 was observed across cell lines and tissue tested. Modified whole cell extracts (1% SDS) (30 µg lysate) of U-2 OS (Lane 1), A-431 (Lane 2), HeLa (Lane 3), Hep G2 (Lane 4), MCF7 (Lane 5), Jurkat (Lane 6) and tissue extract (30 µg lysate) of Mouse Thymus (Lane 7) were electrophoresed using NuPAGE® 4-12 % Bis-Tris gel (Product # NP0321BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # LC2001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:1500 dilution) and detected by chemiluminescence Goat Anti-Rabbit IgG Secondary Antibody, HRP conjugate (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)
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- Experimental details
- MCM6 Polyclonal Antibody detects MCM6 protein at nucleus by immunofluorescent analysis. Sample: U2OS cells were fixed in 4% paraformaldehyde at RT for 15 min. Green: MCM6 protein stained by MCM6 Polyclonal Antibody (Product # PA5-35922) diluted at 1:500. Blue: Hoechst 33342 staining.
- Submitted by
- Invitrogen Antibodies (provider)
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- Experimental details
- Immunofluorescence analysis of MCM6 was performed using 70% confluent PC-3 log phase cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 1 hour at room temperature. The cells were labeled with MCM6 Polyclonal Antibody (Product # PA5-35922) at 1:100 dilution in 0.1% BSA, incubated at 4 degree Celsius overnight and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing nuclear localization. Panel e represents cells with no primary antibody to assess background. The images were captured at 60X magnification..
Supportive validation
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- Invitrogen Antibodies (provider)
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- Experimental details
- MCM6 Polyclonal Antibody detects MCM6 protein at nucleus on human gastric carcinoma by immunohistochemical analysis. Sample: Paraffin-embedded human gastric carcinoma. MCM6 Polyclonal Antibody (Product # PA5-35922) dilution: 1:500. Antigen Retrieval: EDTA based buffer, pH 8.0, 15 min.
- Submitted by
- Invitrogen Antibodies (provider)
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- Experimental details
- MCM6 Polyclonal Antibody detects MCM6 protein at nucleus on human ovarian carcinoma by immunohistochemical analysis. Sample: Paraffin-embedded human ovarian carcinoma. MCM6 Polyclonal Antibody (Product # PA5-35922) dilution: 1:500. Antigen Retrieval: EDTA based buffer, pH 8.0, 15 min.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
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- Experimental details
- Cross-linked ChIP was performed with HeLa chromatin extract and 5 µg of either control rabbit IgG or MCM6 Polyclonal Antibody (Product # PA5-35922). The precipitated DNA was detected by PCR with primer set targeting to Myc promoter.
- Submitted by
- Invitrogen Antibodies (provider)
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- Experimental details
- Enrichment of endogenous MCM6 protein at specific gene loci using Anti-MCM6 Antibody: Chromatin Immunoprecipitation (ChIP) was performed using Anti-MCM6 Rabbit Polyclonal Antibody (Product # PA5-35922, 5 µg) on sheared chromatin from 2 million Hela cells using the MAGnify ChIP System kit (Product # 49-2024). Normal Rabbit IgG was used as a negative IP control. The purified DNA was analyzed by qPCR with PCR primer pairs over cMYC and SAT 2. Data is presented as fold enrichment of the antibody signal versus the negative control IgG using the comparative CT method.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
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- Experimental details
- MCM6 Polyclonal Antibody immunoprecipitates MCM6 protein in IP experiments. IP Sample: 293T whole cell lysate/extract A : 30 µg whole cell lysate/extract of MCM6 protein expressing 293T cells B : Control with 3 µg of pre-immune rabbit IgG C : Immunoprecipitation of MCM6 by 3 µg of MCM6 Polyclonal Antibody (Product # PA5-35922) 7.5% SDS-PAGE The immunoprecipitated MCM6 protein was detected by MCM6 Polyclonal Antibody (Product # PA5-35922) diluted at 1 : 500. Anti-rabbit IgG (HRP) was used as a secondary reagent.