Anti-pan Lamin antibody [X67, X167, X233] (ab20740)

Reviews (2)Q&A (3)References (3)

Overview

  • Product name
    Anti-pan Lamin antibody [X67, X167, X233]
  • Description
    Mouse monoclonal [X67, X167, X233] to pan Lamin
  • Host species
    Mouse
  • Specificity
    Lamins do not appear to be universally distributed among different cell and tissue types. ab20740 recognizes the karyoskeleton, i.e. the intermediate filament proteins of the nucleus. Reacts with Lamin isotypes of Mr 60-75 kDa (LA, LI, LII, LIII, B1, B2, C). Other cell/tissue types have not been tested.
  • Tested applications
    Suitable for: WB, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Cow, Human, Xenopus laevis, Kangaroo, Rainbow trout
  • Immunogen

    cell preparation: Nuclear pore complex-lamina fraction of Xenopus laevis (XLKE-A6 cells).

Properties

Applications

Our Abpromise guarantee covers the use of ab20740 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
WB
ICC/IF
  • Application notes
    ICC/IF: Used at a dilution of 1/50 for 16 hrs (see Abreview for further information). WB: Use at an assay dependent dilution. Predicted molecular weight: 60-75 kDa. Dilution optimised using Chromogenic detection. Not yet tested in other applications. Optimal dilutions/concentrations should be determined by the end user.

    • Target

    Images

    • Immunocytochemistry/ Immunofluorescence - Anti-pan Lamin antibody [X67, X167, X233] (ab20740)
      Immunocytochemistry/ Immunofluorescence - Anti-pan Lamin antibody [X67, X167, X233] (ab20740)

      ab20740 at a 1/50 dilution staining human U2 OS cells by Immunocytochemistry/Immunofluorescence. The cells were Paraformaldehyde fixed and incubated with the antibody for 16 hours. Bound antibody was detected using an Alexa- Fluor ® 488 conjugated goat anti-mouse IgG.

      The antibody was found to stain equally well when used at a dilution of 1/250.

      This image is courtesy of an Abreview submitted by Chi Tang on 6 April 2006.

      See Abreview

    • Immunocytochemistry/ Immunofluorescence - Anti-pan Lamin antibody [X67, X167, X233] (ab20740)
      Immunocytochemistry/ Immunofluorescence - Anti-pan Lamin antibody [X67, X167, X233] (ab20740)Image from Mackay DR et al, J Cell Biol. 2010 Nov 29;191(5):923-31. Epub 2010 Nov 22, Fig 1. DOI 10.1083/jcb.201007124
      ab20740 staining pan Lamin in HeLa cells by Immunocytochemistry/ Immunofluorescence.
      Left image shows pan Lamin.
      Right image shows overlay of pan lamin (green), tubulin (red) and DNA (blue).

    References

    This product has been referenced in:
    • Azevedo AR  et al. Molecular Cytogenetics of Human Single Pronucleated Zygotes. Reprod Sci N/A:N/A (2014). Read more (PubMed: 24717739) »
    • Mackay DR  et al. Defects in nuclear pore assembly lead to activation of an Aurora B-mediated abscission checkpoint. J Cell Biol 191:923-31 (2010). ICC/IF ; Human . Read more (PubMed: 21098116) »
    See all 3 Publications for this product

    Publishing research using ab20740? Please let us know so that we can cite the reference in this datasheet.

    Customer reviews and Q&As

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    1-5 of 5 Abreviews or Q&A

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-pan Lamin antibody [X67, X167, X233]

     Poor
    Abreviews
    Abcam has not validated the combination of species/application used in this Abreview.
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Schmidtea mediterranea Tissue sections (whole animal)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: 95 degree 15min citrate buffer pH6.0
    Permeabilization
    Yes - PBST 15min
    Specification
    whole animal
    Blocking step
    Background Buster as blocking agent for 30 minute(s) · Concentration: 100% · Temperature: 28°C
    Fixative
    Formaldehyde
    Read More

    Abcam user community

    Verified customer

    Submitted Apr 05 2016

    Question

    Will this antibody cross-react with Lamin C? What samples were used to test this antibody in Western blotting?

    Read More
    Answer

    Thank you for your enquiry and patience. This product is a mixture of 3 clones [X67, X167, X233] and two of the clones, X67 and X167, do react with Lamin C. I'm not able to obtain information at this time on exactly what samples were used to test this antibody. Please contact us again if you have any additional questions.

    Read More

    Question

    BATCH NUMBER 185267 ORDER NUMBER seebelow DESCRIPTION OF THE PROBLEM Not correct staining (non-specific staining throughout nucleus and cytoplasm) SAMPLE BJ fibroblasts, Hela cells (cell lines) PRIMARY ANTIBODY Abcam Lamin A antibody [X67, X167, X233] (ab20740) Have tried as suggested on sheet - 16hrs, 1/50 dilution at 4 degrees diluted in 1% BSA Have also tried undiluted, 1/2, and 1/25 with no success washed with PBS POSITIVE AND NEGATIVE CONTROLS USED Tested cells with lamin Ab I know works, and cells were fine. Tried the secondary with another mouse Ab and it worked as expected. Tried Lamin A antibody [X67, X167, X233] with another secondary and it didn't work. ANTIBODY STORAGE CONDITIONS 4 degrees celcius FIXATION OF SAMPLE 4% Formaldehyde for 20 minutes PERMEABILIZATION STEP 0.2% Triton X-100 for 20 minutes BLOCKING CONDITIONS 1% BSA, 20 minutes SECONDARY ANTIBODY Abcam goat anti-mouse IgG at 1:200 diluted in 1% BSA HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 6 HAVE YOU RUN A "NO PRIMARY" CONTROL? Yes DO YOU OBTAIN THE SAME RESULTS EVERY TIME? Yes WHAT STEPS HAVE YOU ALTERED? primary and secondary concentration, secondary ab (used different anti-mouse) ADDITIONAL NOTES purchuse order number wouldn't fit. It is 0300027711

    Read More
    Answer

    Thank you for contacting me regarding the problems that you have experienced using this Ab. Where an Ab is giving non specific staining I would suggest decreasing the concentration at which the primary Ab is used at from 1/50 to 1/100 rather than using the Ab at the higher concentration of ½ and undiluted, the reviewer mentions that using a concentration of 1/250 has been observed to give the same results. I would also advise that you use saponin as a permeabalization reagent (5-10ug/ml) rather than Triton X-100. Triton-X 100 will destroy cellular membranes since the target is located in the nuclear membrane this may be what is resulting in non specific binding, releasing antigen into the cytoplasm. Saponin is a detergent that permeabalizes the cell membrane of eukaryotic cell membranes by interacting with cholesterol and is more gentle than Triton-X. Where membranes are not over treated with saponin they have the ability to reform. In addition to this the percentage of blocking reagent may be increased to 5% BSA this will also act to reduce the observed background staining. Please do not hesitate to get back in touch with me should you still experience problems.

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    Immunocytochemistry/ Immunofluorescence abreview for Anti-pan Lamin antibody [X67, X167, X233]

     Good
    Abreviews
    Abcam guarantees this product to work in the species/application used in this Abreview.
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (U2 OS)
    Specification
    U2 OS
    Fixative
    Paraformaldehyde
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 0.5
    Read More

    Dr. Chi W Tang

    Verified customer

    Submitted Apr 06 2006

    Question

    Do you have information about the epitope recognized by this antibody.

    Read More
    Answer

    Thank you for your enquiry. This product is a mixture of 3 monoclonal antibodies, Clones X67, X167, and X223. The specific epitope locations have not been determined. The reactivity to lamin isotypes in different species for each clone is listed on the datasheet. Please contact us again if you require further assistance.

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

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