Overview

  • Product name

    Anti-MCM7/PRL antibody [47DC141]
    See all MCM7/PRL primary antibodies
  • Description

    Mouse monoclonal [47DC141] to MCM7/PRL
  • Host species

    Mouse
  • Tested applications

    Suitable for: IHC-P, IP, WB, Flow Cyt, ICC/IFmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Dog, Human, Xenopus laevis
  • Immunogen

    Recombinant full length protein corresponding to Human MCM7/PRL.

  • Positive control

    • Breast carcinoma, MAD109 cell lysate, PC12 cell lysate.
  • General notes

     This product was previously labelled as MCM7

     

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Store at -20°C or -80°C. Avoid freeze / thaw cycle.
  • Storage buffer

    Preservative: 0.05% Sodium azide
    Constituent: 1% BSA
  • Concentration information loading...
  • Purity

    IgG fraction
  • Clonality

    Monoclonal
  • Clone number

    47DC141
  • Myeloma

    unknown
  • Isotype

    IgG1
  • Light chain type

    unknown
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab2360 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
IHC-P 1/50 - 1/100. This is when using an ABC method for 30 minutes at room temperature. Sections require high temperature antigen unmasking with 10 mM citrate buffer, pH 6.0 prior to immunostaining.
IP Use at 2 µg/mg of lysate.
WB 1/25 - 1/50. Detects a band of approximately 80 kDa (predicted molecular weight: 80 kDa).

Incubate for 2 hrs at RT for colorimetric detection, can dilute more with ECL+ and with overnight incubation. By Western blot, this antibody detects a band of 80 kDa, which corresponds to the predicted molecular weight of Cdc47 / MCM7/PRL.

Flow Cyt 1/20.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

 

ICC/IF Use a concentration of 1 µg/ml.

Target

  • Function

    Acts as component of the MCM2-7 complex (MCM complex) which is the putative replicative helicase essential for 'once per cell cycle' DNA replication initiation and elongation in eukaryotic cells. The active ATPase sites in the MCM2-7 ring are formed through the interaction surfaces of two neighboring subunits such that a critical structure of a conserved arginine finger motif is provided in trans relative to the ATP-binding site of the Walker A box of the adjacent subunit. The six ATPase active sites, however, are likely to contribute differentially to the complex helicase activity. Required for S-phase checkpoint activation upon UV-induced damage.
  • Sequence similarities

    Belongs to the MCM family.
    Contains 1 MCM domain.
  • Post-translational
    modifications

    Phosphorylated upon DNA damage, probably by ATM or ATR.
  • Cellular localization

    Nucleus.
  • Information by UniProt
  • Database links

  • Alternative names

    • CDABP0042 antibody
    • CDC 47 antibody
    • CDC47 antibody
    • CDC47 homolog antibody
    • Cdc47, S. cerevisiae, homolog of antibody
    • DNA replication licensing factor MCM7 antibody
    • Homolog of S. cerevisiae Cdc47 antibody
    • MCM 2 antibody
    • MCM 7 antibody
    • MCM2 antibody
    • MCM2, formerly antibody
    • Mcm7 antibody
    • MCM7 minichromosome maintenance deficient 7 antibody
    • MCM7_HUMAN antibody
    • Minichromosome Maintainence 7 antibody
    • Minichromosome maintainence, S. cerevisiae, homolog of antibody
    • Minichromosome maintenance complex component 7 antibody
    • Minichromosome maintenance deficient 7 antibody
    • Minichromosome maintenance protein 7 antibody
    • P1.1 MCM3 antibody
    • P1.1-MCM3 antibody
    • P1CDC47 antibody
    • P85MCM antibody
    • PNAS 146 antibody
    • PNAS146 antibody
    • PPP1R104 antibody
    • Protein phosphatase 1, regulatory subunit 104 antibody
    see all

Images

  • All lanes : Anti-MCM7/PRL antibody [47DC141] (ab2360) at 1/200 dilution

    Lane 1 : M phase Xenopus laevis egg extract, whole tissue lysate.
    Lane 2 : I phase Xenopus laevis egg extract, whole tissue lysate.

    Secondary
    All lanes : HRP conjugated Donkey anti-rabbit IgG

    Developed using the ECL technique.

    Predicted band size: 80 kDa
    Observed band size: 95 kDa
    why is the actual band size different from the predicted?


    Exposure time: 1 minute

    See Abreview

  • ab2360 - immunohistochemistry

    Formalin fixed paraffin embedded human tonsil stained with MCM7/PRL, using ABC and DAB chromagen.

  • This image shows immunostaining of MCM7/PRL in rat brain endothelial cells. Brain endothelial cells were co-cultured with neuronal precursor cells and the nuclear staining represents cells in cell cycle. Primary antibody MCM7/PRL (ab2360) was used at 1:50 dilution, incubated overnight at 4 oC.  Secondary antibody - Alexafluor (488 nm) at 1:200 dilution, incubated for 2 hours at room temperature.

    The picture was kindly supplied by Dr Joseph Corteza Lim and Dr Margery Barrand from University of Cambridge, Department of Pharmacology.

     
     
  • ab2360 staining MCM7/PRL in human bladder cancer tissue sections by Immunohistochemistry (formalin fixed sections). Tissue was fixed with formaldehyde and a heat mediated antigen retrieval step was performed using citrate buffer. Tissue was blocked with 5% BSA for 1 hour at room temperature followed by incubation with the primary antibody at a 1/1200 dilution for 1 hour. A HRP-conjugated goat anti-mouse polyclonal was used as secondary antibody un-diluted.

    See Abreview

  • Lane 1 : Anti-MCM7/PRL antibody [47DC141] (ab2360) at 1/50 dilution
    Lane 2 : Anti-MCM7/PRL antibody [47DC141] (ab2360) at 1/200 dilution
    Lane 3 : Anti-MCM7/PRL antibody [47DC141] (ab2360) at 1/500 dilution

    All lanes : Whole cell lysate prepared from SW780 cells

    Lysates/proteins at 25 µg per lane.

    Secondary
    All lanes : Goat anti-mouse IgG conjugated to HRP at 1/5000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 80 kDa
    Observed band size: 81 kDa why is the actual band size different from the predicted?


    Exposure time: 10 minutes


    Gel run under denaturing conditions 4-12% gradient.

    See Abreview

  • ICC/IF image of MCM7/PRL (ab2360) stained HeLa cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab2360, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

  • Overlay histogram showing HeLA cells stained with MCM7/PRL (ab2360) (red line). The cells were fixed with 100% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab2360, 1/20 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.

  • ab2360 staining MCM7/PRL in formalin-fixed, paraffin-embedded Human breast carcinoma tissue tissue by Immunohistochemistry.

References

This product has been referenced in:

  • Kovary KM  et al. Expression variation and covariation impair analog and enable binary signaling control. Mol Syst Biol 14:e7997 (2018). Read more (PubMed: 29759982) »
  • Drissi R  et al. Destabilization of the MiniChromosome Maintenance (MCM) complex modulates the cellular response to DNA double strand breaks. Cell Cycle 17:2593-2609 (2018). Read more (PubMed: 30516086) »
See all 22 Publications for this product

Customer reviews and Q&As

1-7 of 7 Abreviews or Q&A

Application
Western blot
Sample
Human Cell lysate - whole cell (293T)
Gel Running Conditions
Non-reduced Non-Denaturing (Native)
Loading amount
60 µg
Treatment
MMC 2H
Specification
293T
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 37°C

Abcam user community

Verified customer

Submitted Sep 19 2018

Application
Western blot
Sample
Mouse Cell lysate - whole cell (3T3MEF)
Gel Running Conditions
Non-reduced Denaturing (7.5% acrylamide-Bis gel)
Loading amount
50000 cells
Treatment
by serum starvation for 48h , cells are synchronized at G0 phase and released with 5% FBS and collected at indicated time
Specification
3T3MEF
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 5%

Abcam user community

Verified customer

Submitted Dec 18 2015

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (Tonsil)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: TRIS-EDTA-Buffer pH 9,0
Permeabilization
Yes - with wash - Buffer with Tween
Specification
Tonsil
Fixative
Paraformaldehyde

Mr. Rudolf Jung

Verified customer

Submitted Jun 10 2013

Application
Western blot
Sample
Human Cell lysate - whole cell (SW780 bladder cancer)
Gel Running Conditions
Reduced Denaturing (4-12%)
Loading amount
25 µg
Specification
SW780 bladder cancer
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 19°C

Dr. Karin Birkenkamp-Demtroeder

Verified customer

Submitted May 06 2010

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (Bladder Cancer / Transitional cell Carcinoma)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citrate buffer
Permeabilization
No
Specification
Bladder Cancer / Transitional cell Carcinoma
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
Fixative
Formaldehyde

Dr. Karin Birkenkamp-Demtroeder

Verified customer

Submitted Apr 29 2010

Application
Western blot
Sample
Human Cell lysate - whole cell (293T (LANE 1 AND 2))
Gel Running Conditions
Reduced Denaturing (10%)
Loading amount
25 µg
Specification
293T (LANE 1 AND 2)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Dec 31 2008

Application
Western blot
Sample
Xenopus laevis Tissue lysate - whole (Egg extracts)
Loading amount
12 µg
Specification
Egg extracts
Blocking step
Milk as blocking agent for 20 minute(s) · Concentration: 5%

Abcam user community

Verified customer

Submitted Jul 25 2006

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