Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-Lamin A + Lamin C antibody [WL4G10] - BSA and Azide free (ab269575)

Key features and details

  • Mouse monoclonal [WL4G10] to Lamin A + Lamin C - BSA and Azide free
  • Suitable for: ICC/IF, IHC-P, WB
  • Knockout validated
  • Reacts with: Human
  • Isotype: IgG1

Overview

  • Product name

    Anti-Lamin A + Lamin C antibody [WL4G10] - BSA and Azide free
    See all Lamin A + Lamin C primary antibodies

  • Description

    Mouse monoclonal [WL4G10] to Lamin A + Lamin C - BSA and Azide free

  • Host species

    Mouse

  • Tested applications

    Suitable for: ICC/IF, IHC-P, WBmore details

  • Species reactivity

    Reacts with: Human

  • Immunogen

    Recombinant fragment (GST-tag) corresponding to Human Lamin A + Lamin C (C terminal). (C-terminal fragment of Lamin A)
    Database link: P02545

  • Positive control

    • ICC/IF: HAP1 wild-type and HAP1-LMNA knockout cells, HeLa cells. IHC-P: FFPE human normal skin WB: HAP1 and HeLa whole cell lysates

  • General notes

    This antibody clone is manufactured by Abcam. If you require a custom buffer formulation or conjugation for your experiments, please contact orders@abcam.com.

    ab269575 is the carrier-free version of ab232730. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with

    This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

    Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

    Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

    We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

    In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

    We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

    Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

    Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.

Properties

Images

  • Western blot - Anti-Lamin A + Lamin C antibody [WL4G10] - BSA and Azide free (ab269575)
    Western blot - Anti-Lamin A + Lamin C antibody [WL4G10] - BSA and Azide free (ab269575)
    All lanes : Anti-Lamin A + Lamin C antibody [WL4G10] (ab232730) at 1/1000 dilution

    Lane 1 : Wild-type HeLa cell lysate
    Lane 2 : Lamin A + C knockout HeLa cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 74 kDa



    This data was developed using the same antibody clone in a different buffer formulation (ab232730).

    Lanes 1-2: Merged signal (red and green). Green - ab232730 observed at 70,75 kDa. Red - loading control ab181602 observed at 37 kDa.

     ab232730 Anti-Lamin A + C antibody [WL4G10] was shown to specifically react with Lamin A + C in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab261787 (knockout cell lysate ab256979) was used. Wild-type and Lamin A + C knockout samples were subjected to SDS-PAGE. ab232730 and Anti-GAPDH antibody[EPR16891] - Loading Control (ab181602) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (ab216772) and Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (ab216777) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

     

     

  • Immunocytochemistry/ Immunofluorescence - Anti-Lamin A + Lamin C antibody [WL4G10] - BSA and Azide free (ab269575)
    Immunocytochemistry/ Immunofluorescence - Anti-Lamin A + Lamin C antibody [WL4G10] - BSA and Azide free (ab269575)

    ab232730 staining Lamin A+C in HeLa (Human epithelial cell line from cervix adenocarcinoma) cells.

    The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% PBS-Triton X-100 for 5 mins and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab232730 at 1 μg/ml and ab6046, Rabbit polyclonal to beta Tubulin - Loading Control, at 1/1000 dilution. Cells were then incubated with ab150117, Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) at 1/1000 dilution (shown in green) and ab150084, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 594) at 1/1000 dilution (shown in pseudocolor red). Nuclear DNA was labeled with DAPI (shown in blue).

    This image was produced using the same antibody clone but in a different formulation ab232730, PBS and sodium azide.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Lamin A + Lamin C antibody [WL4G10] - BSA and Azide free (ab269575)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Lamin A + Lamin C antibody [WL4G10] - BSA and Azide free (ab269575)

    IHC image of Lamin A + C staining in a section of formalin-fixed paraffin-embedded normal human skin* performed on a Leica BONDTM system using the standard protocol F.

    The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab232730, 1 µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with hematoxylin and mounted with DPX. The inset secondary-only control image is taken from an identical assay without primary antibody.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

    *Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre

    This image was produced using the same antibody clone but in a different formulation ab232730, PBS and sodium azide.

  • Western blot - Anti-Lamin A + Lamin C antibody [WL4G10] - BSA and Azide free (ab269575)
    Western blot - Anti-Lamin A + Lamin C antibody [WL4G10] - BSA and Azide free (ab269575)
    All lanes :

    Lane 1 : HAP1 whole cell lysate
    Lane 2 : HAP1 LMNA KO whole cell lysate
    Lane 3 : HeLa whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 74 kDa



    This image was produced using the same antibody clone but in a different formulation ab232730, PBS and sodium azide.

    ab232730 was shown to specifically react with LMNA (Lamin A + C) in wild type HAP1 cells. No band was observed when LMNA (Lamin A + C) knockout samples were used. Wild-type and LMNA (Lamin A + C) knockout samples were subjected to SDS-PAGE. ab232730 and ab181602 (Rabbit anti GAPDH) were incubated overnight at 4°C at a 1mg/ml concentration and 1/10000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

     

  • Immunocytochemistry/ Immunofluorescence - Anti-Lamin A + Lamin C antibody [WL4G10] - BSA and Azide free (ab269575)
    Immunocytochemistry/ Immunofluorescence - Anti-Lamin A + Lamin C antibody [WL4G10] - BSA and Azide free (ab269575)

    ab232730 staining Lamin A+C (colored green) in wild-type HAP1 cells (top panel) and LMNA knockout HAP1 cells (bottom panel).

    The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% Triton X-100 for 5 mins and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab232730 at 1 μg/ml and ab6046, Rabbit polyclonal to beta Tubulin - Loading Control, at 1/1000 dilution overnight at 4ºC. Cells were then incubated with ab150117, Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) at 1/1000 dilution (shown in green) and ab150084, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 594) at 1/1000 dilution (shown in pseudocolor red). Nuclear DNA was labeled with DAPI (shown in blue).

    Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

    This image was produced using the same antibody clone but in a different formulation ab232730, PBS and sodium azide.

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