Overview

  • Product name
    Anti-DMC1 antibody [2H12/4]
    See all DMC1 primary antibodies
  • Description
    Mouse monoclonal [2H12/4] to DMC1
  • Host species
    Mouse
  • Specificity
    Specific for DMC1 - does not cross-react with the related protein Rad51. The antibody was tested on mouse meiotic chromosomes. It allowed localisation of the Dmc1 protein as numerous foci in the Leptotene and Zygotene stages of meiotic prophase, and fewer foci in the early Pachytene stage of meiotic prophase.
  • Tested applications
    Suitable for: Flow Cyt, IP, WB, ICC/IF, IHC-Pmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Cow, Human
  • Immunogen

    Recombinant fusion protein (Human).

  • Positive control
    • ICC:mouse meiotic chromosomes

Properties

  • Form
    Liquid
  • Storage instructions
    Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
  • Storage buffer
    Constituent: PBS
  • Concentration information loading...
  • Purity
    Protein A purified
  • Clonality
    Monoclonal
  • Clone number
    2H12/4
  • Myeloma
    Sp2/0-Ag14
  • Isotype
    IgG2a
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab11054 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
Flow Cyt Use 1µg for 106 cells.

ab170191 - Mouse monoclonal IgG2a, is suitable for use as an isotype control with this antibody.

 

IP Use at an assay dependent concentration.
WB Use at an assay dependent concentration. Predicted molecular weight: 41 kDa.
ICC/IF 1/100. Fix cells using a 4% paraformaldehyde solution and permeabilize using SDS (0.03% in a 4% paraformaldehyde solution). Incubate antibody overnight.
IHC-P Use at an assay dependent concentration.

Target

  • Function
    May participate in meiotic recombination, specifically in homologous strand assimilation, which is required for the resolution of meiotic double-strand breaks.
  • Sequence similarities
    Belongs to the RecA family. DMC1 subfamily.
    Contains 1 HhH domain.
  • Cellular localization
    Nucleus. Chromosome.
  • Information by UniProt
  • Database links
  • Alternative names
    • Disrupted meiotic cDNA 1 homolog antibody
    • Disrupted meiotic cDNA 1, yeast, homolog of antibody
    • dJ199H16.1 antibody
    • DMC 1 antibody
    • DMC1 antibody
    • DMC1 dosage suppressor of mck1 homolog antibody
    • DMC1 dosage suppressor of mck1 homolog meiosis specific homologous recombination (yeast) antibody
    • DMC1 homologue antibody
    • DMC1_HUMAN antibody
    • DMC1H antibody
    • DNA meiotic recombinase 1 antibody
    • HsLim15 antibody
    • LIM15 antibody
    • Meiotic recombination protein DMC1/LIM15 homolog antibody
    • MGC150472 antibody
    • MGC150473 antibody
    see all

Images

  • Overlay histogram showing HeLa cells stained with ab11054 (red line). The cells were fixed with 80% 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 (ab11054, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG2a [ICIGG2A] (ab91361, 1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.

References

This product has been referenced in:
  • Hopkins J  et al. Meiosis-specific cohesin component, Stag3 is essential for maintaining centromere chromatid cohesion, and required for DNA repair and synapsis between homologous chromosomes. PLoS Genet 10:e1004413 (2014). ICC/IF ; Mouse . Read more (PubMed: 24992337) »
  • Sobinoff AP  et al. Damaging legacy: maternal cigarette smoking has long-term consequences for male offspring fertility. Hum Reprod 29:2719-35 (2014). IHC ; Mouse . Read more (PubMed: 25269568) »
See all 7 Publications for this product

Customer reviews and Q&As

1-10 of 11 Abreviews or Q&A

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (HeLa)
Specification
HeLa
Permeabilization
Yes - 0.5% Triton X100 in PBS
Fixative
Paraformaldehyde

Dr. Kirk Mcmanus

Verified customer

Submitted Nov 15 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (adult testes)
Specification
adult testes
Fixative
4% PFA
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: 0.01M citric acid/citrate sodium
Permeabilization
No
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Dr. Qing Wen

Verified customer

Submitted Mar 18 2013

Answer

Thank you for your enquiry.

I am sorry to hear the replacement antibody is still not working as expected. I would be happy to send you different Dmc1 antibody of your choice. So I can process your request, please include the purchase order number or Abcam order reference number so I can locate your order on our system.

Thanks in advance for the additional information.

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Question
Answer

Merci de nous avoir contactés. Nous sommes désolés d'apprendre que le produit que vous avez reçu ne fonctionne pas comme attendu. Comme convenu j'ai mis en place l'envoi d'une unité de remplacement avec un lot différent, le numéro de commande de remplacement gratuit de ce produit : ******. Vous recevrez prochainement un mail de confirmation comprenant les détails d'expédition. N'hésitez pas à nous contacter lors d'une prochaine occasion.

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Answer

Thank you for contacting us and I'm sorry to hear that this antibody is giving you some trouble. I think that boiling will more properly denature the protein and allow it to move more efficiently through the gel. To denature, use a loading buffer with SDS and boil the mixture at 95-100°C for 5 minutes. You can find our general WB protocol here: https://www.abcam.com/ps/pdf/protocols/WB-beginner.pdf Hopefully this will improve the resolution of the band, but it probably wouldn't hurt to do a nuclear fractionation to isolate the protein of interest. Our protocol is specific for cells: https://www.abcam.com/index.html?pageconfig=resource&rid=11408 but there is some literature on isolation of nuclei from tissues: http://www.ncbi.nlm.nih.gov/pubmed/18228353 I don't think you'll need it, but I just wanted to make you aware that we do carry a new nuclear extraction kit for cells and tissues that was just added to our catalogue yesterday: https://www.abcam.com/EpiSeeker-Nuclear-Extraction-Kit-ab113474.html I hope this information helps. Please contact us with any other questions.

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Question
Answer

I'm very sorry to hear you are still having no staining with the recent set of experiments, I appreciate the effort you have put to make the experiments work and cannot understand why the antibody did not work for you. We have not received any other complaints on this product in a year so I suspect your vial was damaged during shipping or storage. I would be happy to offer you a refund, credit note or replacement vial if you can tell me which you would prefer. I look forward to hearing from you to arrange this,

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Answer

I am pleased to inform you that I have received just this morning a detailed protocol for staining with ab11054. The antibody has been very satisfactory in detecting the Dmc1 protein on mouse meiotic chromosomes. The meiotic chromosomes spreads were performed as described in the protocol that I have added to the online datasheet (accessible via the link below). Which means cells were fixed using a 4% paraformaldehyde, solution and permeabilized using SDS (0.03% in a 4% paraformaldehyde solution). The IF localization of Dmc1 was then conducted as described in the attached protocol. The 2H12/4 antibody was diluted 1/100 In 500µl of 1X ADB and incubated overnight at 4ºC. The secondary antibody used was a Goat anti-Mouse antibody labeled with Alexa-488 dye (A11017, Invitrogen), diluted 1/500. The slides were then mounted in a drop of Vectashield mounting medium with DAPI (H-1500, Vector Laboratories). This procedure allowed localization of the Dmc1 protein as numerous foci in the Leptotene and Zygotene stages of meiotic prophase, and fewer foci in the early Pachytene stage of meiotic prophase. I hope this information will be useful, please let me know if you still experience problems,

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Answer

I'm very sorry for the delay in finding out more information to help you and to hear you are experiencing problems with two of our antibodies. I have been informed that ab36453 has been used in the following reference: Geddis AE & Kaushansky K. Megakaryocytes express functional Aurora-B kinase in endomitosis. Blood 104:1017-24 (2004). PubMed: 15130946 I enclose the staining protocol for this antibody: ICC/IF: 1/1000. Fix cells in suspension by the addition of an equal volume of 4% paraformaldehyde prepared in phosphate-buffered saline (PBS, pH 7.4) for a final concentration of 2% paraformaldehyde. Fix cells at room temperature for 45 minutes and then centrifuge and wash twice with PBS. Resuspend cells in PBST (PBS containing 0.5% Triton-X100) with 3% bovine serum albumin (BSA) and incubate at room temperature for 15 to 30 minutes to provide blocking against nonspecific antibody reactivity. Unfortunately I am still trying to find out the staining protocol with ab11054, I apologize for the delay. I would like to offer for me to look at your protocol as I may be able to make suggestions to improve the staining. I enclose the link below which will take you to a short questionnaire. If you please put it to my attention I will be able to continue my investigations and hope to help you resolve this problem: https://www.abcam.com/index.html?section=ihc&pageconfig=technical&intAbID=11054&mode=questionaire I look forward to hearing from you and hope the protocol for ab36453 will be useful,

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Mouse Tissue lysate - whole (testis)
Loading amount
40 µg
Specification
testis
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5%

Abcam user community

Verified customer

Submitted May 25 2006

Answer

Thank you for your enquiry. I am sorry to hear that your customer has been having difficulties with this antibody. Quality is important to Abcam. Your customer is using a western blotting approach that I would largely recommend. I would be very interested to know whether this cross reacting product is the major product on the blot or whether this is a mild cross reacting band. I would therefore appreciate it if you could ask your customer if they could -mail me a blot image as this would be most informative. We take all complaints very seriously and if there is a major cross reacting band we need to consider to remove it from our catalogue. I look forward to hearing from you.

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1-10 of 11 Abreviews or Q&A

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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