PA5-29039
antibody from Invitrogen Antibodies
Targeting: MCM4
CDC21, CDC54, hCdc21, MGC33310, P1-Cdc21
Antibody data
- Antibody Data
- Antigen structure
- References [0]
- Comments [0]
- Validations
- Immunocytochemistry [3]
- Immunoprecipitation [1]
- Immunohistochemistry [1]
- Chromatin Immunoprecipitation [5]
- Other assay [1]
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Validation data
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- Product number
- PA5-29039 - Provider product page
- 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.
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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.
- 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.
- 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 (Heavy Chain) 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.
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.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemical analysis of paraffin-embedded A549 Xenograft, using MCM4 (Product # PA5-29039) 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 a MCM4 polyclonal antibody (Product # PA5-29039). The precipitated DNA was detected by PCR with primer set targeting to Myc promoter.
- 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.
- 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.
- 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.
- 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
- 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.