MA5-15923

antibody from Invitrogen Antibodies

Targeting: MCM2 BM28, CCNL1, cdc19, CDCL1, D3S3194, DFNA70, KIAA0030

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Immunohistochemistry (Supportive data in Antibodypedia)

Flow cytometry (Supportive data in Antibodypedia)

Antibody Data

Product number

MA5-15923

Provider

Invitrogen Antibodies

Product name

MCM2 Monoclonal Antibody (1E7)

Antibody type

Monoclonal

Antigen

Purifed from natural sources

Description

MA5-15923 targets MCM2 in FACS, IF, IHC, and WB applications and shows reactivity with Human samples.

Antibody clone number

1E7

Concentration

Conc. Not Determined

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis of MCM2 using MCM2 monoclonal antibody (Product # MA5-15923) in MCF-7 (1), HeLa (2), Jurkat (3), K562 (4), HEK293 (5) and HEPG2 (6) cell lysate.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Knockdown of MCM2 was achieved by transfecting MCF7 with MCM2 specific siRNAs (Silencer® select Product # s8586, s8587). Western blot analysis (Fig. a) was performed using Nuclear enriched extracts from the MCM2 untransfected cells (lane 1), non-targeting scrambled siRNA transfected cells (lane 2) and knockdown cells (lane 3). The blot was probed with MCM2 Monoclonal Antibody (1E7) (Product # MA5-15923, 1:1000 dilution ) and Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 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 MCM2.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot was performed using Anti-MCM2 Monoclonal Antibody (Product # MA5-15923) and a 125 kDa band corresponding to MCM2 was observed across all cell lines. Expression of MCM2 was observed to be downregulated upon treatment of LNCaP and PC-3 cells with Trichostatin A (100 ng/mL for 24 hours) (DOI: 10.1158/0008-5472.CAN-09-4176). Modified whole cell extracts (1% SDS) (50 µg lysate) of LNCaP (Lane 1), LNCaP treated with Trichostatin A (100 ng/mL for 24 hours) (Lane 2), PC-3 (Lane 3) and PC-3 treated with Trichostatin A (100 ng/mL for 24 hours) (Lane 4) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein 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-Mouse IgG (H+L), Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Reagent Kit (Product # WP20005).

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot was performed using Anti-MCM2 Monoclonal Antibody (1E7)(Product # MA5-15923) and a 125kDa band corresponding to MCM2 was observed across cell lines tested. Nuclear enriched extracts (30 µg lysate) of MCF7 (Lane 1), HeLa (Lane 2), A549 (Lane 3), K-562 (Lane 4), Jurkat (Lane 5), MOLT-4 (Lane 6) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein 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-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177,1:4000 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

Western blot analysis of MCM2 using a MCM2 monoclonal antibody (Product # MA5-15923) against a human MCM2 (AA: 16-232) recombinant protein.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis of MCM2 using MCM2 monoclonal antibody (Product # MA5-15923) in MCF-7 (1), HeLa (2), Jurkat (3), K562 (4), HEK293 (5) and HEPG2 (6) cell lysate.

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of HeLa cells using MCM2 monoclonal antibody (Product # MA5-15923) (Green). Blue: DRAQ5 fluorescent DNA dye. Red: actin filaments have been labeled with phalloidin.

Immunohistochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunohistochemical analysis of paraffin-embedded ovarian cancer tissues using MCM2 monoclonal antibody (Product # MA5-15923) followed with DAB staining.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunohistochemical analysis of paraffin-embedded colon cancer tissues using MCM2 monoclonal antibody (Product # MA5-15923) followed with DAB staining.

Flow cytometry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Flow cytometric analysis of Jurkat cells using MCM2 monoclonal antibody (Product # MA5-15923) (green) and negative control (red).