PA5-29039

antibody from Invitrogen Antibodies

Targeting: MCM4 CDC21, CDC54, hCdc21, MGC33310, P1-Cdc21

Western blot (Data presented on provider website)

Immunocytochemistry (Supportive data in Antibodypedia)

Immunoprecipitation (Data presented on provider website)

Immunohistochemistry (Data presented on provider website)

Chromatin Immunoprecipitation (Supportive data in Antibodypedia)

Other assay (Supportive data in Antibodypedia)

Antibody Data

Product number

PA5-29039

Provider

Invitrogen Antibodies

Product name

MCM4 Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Recombinant protein fragment

Description

Recommended positive controls: Jurkat, Raji, NCI-H929, Neuro 2A, C8D30, NIH-3T3, Raw264.7, C2C12. Predicted reactivity: Mouse (96%), Rat (95%), Zebrafish (88%), Xenopus laevis (89%), Rhesus Monkey (99%), Bovine (93%). Store product as a concentrated solution. Centrifuge briefly prior to opening the vial.

Reactivity

Human, Mouse

Host

Rabbit

Isotype

IgG

Vial size

100 µL

Concentration

1 mg/mL

Storage

Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescent analysis of MCM4 in paraformaldehyde-fixed HeLa cells using a MCM4 polyclonal antibody (Product # PA5-29039) at a 1:500 dilution.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of MCM4 was performed using 70% confluent log phase SH-SY5Y 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 MCM4 Polyclonal Antibody (Product # PA5-29039) at 5 µg/mL in 0.1% BSA, incubated at 4 degree Celsius overnight and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant 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 SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing nuclear localization. Panel f represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Chromatin Immunoprecipitation

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 a MCM4 polyclonal antibody (Product # PA5-29039). 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

Cross-linked ChIP was performed with HeLa chromatin extract and 5 µg of either control rabbit IgG or MCM4 Polyclonal Antibody (Product # PA5-29039). 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

Chromatin Immunoprecipitation (ChIP) assay of endogenous MCM4 protein using Anti-MCM4 Antibody: ChIP was performed using Anti-MCM4 Rabbit Polyclonal Antibody (Product # PA5-29039) 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 transcriptional start site and gene body (+2kb) of GAPDH, transcriptional start site of MYC and SAT2 satellite repeats. Data is presented as fold enrichment of the antibody signal versus the negative control IgG using the comparative CT method.

Other assay

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

MCM4 Polyclonal Antibody immunoprecipitates MCM4 protein in IP experiments. IP samples: Jurkat whole cell extract. A. 40 µg Jurkat whole cell extract. B. Control with 4 µg of preimmune Rabbit IgG. C. Immunoprecipitation of MCM4 protein by 4 µg MCM4 Polyclonal Antibody (Product # PA5-29039). 7.5 % SDS-PAGE. The immunoprecipitated MCM4 protein was detected by MCM4 Polyclonal Antibody (Product # PA5-29039) diluted at 1:500.