PA5-22104
antibody from Invitrogen Antibodies
Targeting: MCM7
CDC47, MCM2, PPP1R104
Antibody data
- Antibody Data
- Antigen structure
- References [0]
- Comments [0]
- Validations
- Western blot [3]
- Immunocytochemistry [2]
- Immunohistochemistry [1]
- Chromatin Immunoprecipitation [2]
- Other assay [1]
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Validation data
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- Product number
- PA5-22104 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- MCM7 Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Recombinant protein fragment
- Description
- Recommended positive controls: 293T, A431, H1299, HeLa, HepG2, Molt-4, Raji. Predicted reactivity: Mouse (92%), Rat (93%), Zebrafish (85%), Xenopus laevis (88%), Bovine (98%). Store product as a concentrated solution. Centrifuge briefly prior to opening the vial.
- Reactivity
- Human
- Host
- Rabbit
- Isotype
- IgG
- Vial size
- 100 µL
- Concentration
- 0.76 mg/mL
- Storage
- Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western Blot using MCM7 Polyclonal Antibody (Product # PA5-22104). Sample (30 µg of whole cell lysate). Lane A: H1299. 7.5% SDS PAGE. MCM7 Polyclonal Antibody (Product # PA5-22104) 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
- Knockdown of MCM7 was achieved by transfecting HeLa with MCM7 specific siRNAs (Silencer® select Product # s8602, s8603). Western blot analysis (Fig. a) was performed using modified whole cell extracts (1% SDS) from the MCM7 knockdown cells (Lane 3), non-specific scrambled siRNA transfected cells (Lane 2) and untransfected cells (Lane 1). The blot was probed with MCM7 Polyclonal Antibody (Product # PA5-22104, 1:1000 dilution) and Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000 dilution). Densitometric analysis of this western blot is shown in histogram (Fig. b). Loss of signal upon siRNA mediated knock down confirms that antibody is specific to MCM7. An uncharacterized band was observed at ~45kDa.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Anti-MCM7 Polyclonal Antibody (Product # PA5-22104) and a 80kDa band corresponding to MCM7 was observed in all the tested cell models along with an uncharacterized band at ~35kDa. Modified whole cell extracts (1% SDS) (30ug lysate) of PC-3 (Lane 1), LNCaP (Lane 2), DU 145 (Lane 3), HeLa (Lane 4), MCF7 (Lane 5), MKN45 (Lane 6), SW480 (Lane 7), and Jurkat (Lane 8) were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris gel (Product # NP0322BOX). 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 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)
- Main image
- Experimental details
- Immunofluorescent analysis of MCM7 in paraformaldehyde-fixed HeLa cells using a MCM7 polyclonal antibody (Product # PA5-22104) at a 1:200 dilution.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescence analysis of paraformaldehyde-fixed HeLa, using MCM7 antibody (Product # PA5-22104) at 1:200 dilution.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemical analysis of paraffin-embedded Cal27 Xenograft, using MCM7 (Product # PA5-22104) antibody at 1:100 dilution. 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 MCM7 Polyclonal Antibody (Product # PA5-22104). The precipitated DNA was detected by PCR with primer set targeting to Myc promoter.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Chromatin Immunoprecipitation (ChIP) assay of endogenous MCM7 protein using Anti-MCM7 Antibody: ChIP was performed using Anti-MCM7 Rabbit Polyclonal Antibody (Product # PA5-22104, 5 µg) on sheared chromatin from 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 using primers binding to promoter and transcriptional start site of MYC, AURKA transcriptional start site and SAT2 satellite repeats. 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)
- Main image
- Experimental details
- MCM7 Polyclonal Antibody immunoprecipitates MCM7 protein in IP experiments. IP samples: HeLa whole cell extract. A. 40 µg HeLa whole cell extract. B. Control with 4 µg of preimmune Rabbit IgG. C. Immunoprecipitation of MCM7 protein by 4 µg MCM7 Polyclonal Antibody (Product # PA5-22104). 7.5 % SDS-PAGE. The immunoprecipitated MCM7 protein was detected by MCM7 Polyclonal Antibody (Product # PA5-22104) diluted at 1:1,000.