Anti-Lamin B2 antibody [LN43] (ab8983)

Reviews (5)Q&A (7)References (27)

Overview

  • Product name

    Anti-Lamin B2 antibody [LN43]
    See all Lamin B2 primary antibodies

  • Description

    Mouse monoclonal [LN43] to Lamin B2

  • Host species

    Mouse

  • Specificity

    This antibody reacts with an epitope located in the C-terminal part of lamin B2. Lamins do not appear to be universally distributed among different cell and tissue types. Please see the listed positive controls for this antibody. Other cell/tissue types have not been tested.

  • Tested applications

    Suitable for: ICC, Flow Cyt, IHC-Fr, WB, IP, ICC/IFmore details

  • Species reactivity

    Reacts with: Mouse, Hamster, Human, Xenopus laevis, Zebrafish

  • Immunogen

    Tissue, cells or virus corresponding to Lamin B2. Immunogen = detergent insoluble fraction of potoroo cell line PtK1.

  • Epitope

    Epitope located in the C-terminal part of lamin B2.

  • Positive control

    • IHC-Fr: testicular germ cell tumour sections (see Machiels et al 1997), parabasal cells of skin epidermis (see Broers et al). WB: NCI-H125 nuclear cell lysate Jurkat cells.

Properties

Applications

Our Abpromise guarantee covers the use of ab8983 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
ICC 1/400.
Flow Cyt Use 1µg for 106 cells.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
IHC-Fr 1/1.

(when used with an avidin-biotinylated HRP complex as detection agent).
WB 1/100 - 1/1000. Detects a band of approximately 65-67 kDa.
IP Use at an assay dependent concentration.
ICC/IF Use at an assay dependent concentration. PubMed: 22403714

Target

  • Function

    Lamins are components of the nuclear lamina, a fibrous layer on the nucleoplasmic side of the inner nuclear membrane, which is thought to provide a framework for the nuclear envelope and may also interact with chromatin.

  • Involvement in disease

    Defects in LMNB2 are a cause of partial acquired lipodystrophy (APLD) [MIM:608709]. A rare childhood disease characterized by loss of subcutaneous fat from the face and trunk. Fat deposition on the pelvic girdle and lower limbs is normal or excessive. Most frequently, onset between 5 and 15 years of age. Most affected subjects are females and some show no other abnormality, but many develop glomerulonephritis, diabetes mellitus, hyperlipidemia, and complement deficiency. Mental retardation in some cases. APLD is a sporadic disorder of unknown etiology.

  • Sequence similarities

    Belongs to the intermediate filament family.

  • Post-translational
    modifications

    B-type lamins undergo a series of modifications, such as farnesylation and phosphorylation. Increased phosphorylation of the lamins occurs before envelope disintegration and probably plays a role in regulating lamin associations.

  • Cellular localization

    Nucleus inner membrane.
  • Information by UniProt

  • Database links

    • Alternative names

      • LAMB 2 antibody
      • LAMB2 antibody
      • Lamin-B2 antibody
      • LMN 2 antibody
      • LMN B2 antibody
      • LMN2 antibody
      • LMNB 2 antibody
      • LMNB2 antibody
      • LMNB2_HUMAN antibody
      • MGC2721 antibody
      • RGD1563803 antibody
      see all

    Images

    • Western blot - Anti-Lamin B2 antibody [LN43] (ab8983)
      Western blot - Anti-Lamin B2 antibody [LN43] (ab8983)
      All lanes : Anti-Lamin B2 antibody [LN43] (ab8983) at 1 µg/ml

      Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
      Lane 2 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate
      Lane 3 : U2OS (Human osteosarcoma cell line) Whole Cell Lysate
      Lane 4 : HL60 (Human promyelocytic leukemia cell line) Whole Cell Lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilution

      Developed using the ECL technique.

      Performed under reducing conditions.

      Observed band size: 75 kDa
      why is the actual band size different from the predicted?
      Additional bands at: 37 kDa. We are unsure as to the identity of these extra bands.


      Exposure time: 20 minutes
    • Immunohistochemistry (Frozen sections) - Anti-Lamin B2 antibody [LN43] (ab8983)
      Immunohistochemistry (Frozen sections) - Anti-Lamin B2 antibody [LN43] (ab8983)

      IHC-Fr of human kidney showing nuclear lamina staining in the ductal epithelium.  

    • Immunocytochemistry/ Immunofluorescence - Anti-Lamin B2 antibody [LN43] (ab8983)
      Immunocytochemistry/ Immunofluorescence - Anti-Lamin B2 antibody [LN43] (ab8983)Image from Tsai YJ et al., PLoS One. 2012;7(3):e32820. Epub 2012 Mar 5. Fig 5.; doi:10.1371/journal.pone.0032820; March 5, 2012, PLoS ONE 7(3): e32820.
      Immunofluorescence analysis of HeLa cells in telophase, staining Lamin B2 with ab8983.
    • Immunocytochemistry/ Immunofluorescence - Anti-Lamin B2 antibody [LN43] (ab8983)
      Immunocytochemistry/ Immunofluorescence - Anti-Lamin B2 antibody [LN43] (ab8983)This image is courtesy of Marilena Ciciarello & Patrizia Lavia, University La Sapienza

      HeLa cells were fixed in 100% methanol for 6 minutes at -20°C then stained with ab8983 (1/400). This image shows that ab8983 clearly stains the nuclear envelope (green). The DNA is stained with DAPI (blue).

    • Immunoprecipitation - Anti-Lamin B2 antibody [LN43] (ab8983)
      Immunoprecipitation - Anti-Lamin B2 antibody [LN43] (ab8983)
      Lamin B2 - Nuclear Envelope Marker was immunoprecipitated using 0.5mg Hela whole cell extract, 5µg of Mouse monoclonal to Lamin B2 - Nuclear Envelope Marker (ab8983) and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
      The antibody was incubated under agitation with Protein G beads for 10min, Hela whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
      Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab8983.
      Secondary: Goat polyclonal to mouse IgG light chain specific (HRP) at 1/5000 dilution.
      Band: 73kDa: Lamin B2
    • Flow Cytometry - Anti-Lamin B2 antibody [LN43] (ab8983)
      Flow Cytometry - Anti-Lamin B2 antibody [LN43] (ab8983)

      Overlay histogram showing HeLA cells stained with ab8983 (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 (ab7481) / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab8983, 1µg/1x106 cells) 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.

    • Immunohistochemistry (Frozen sections) - Anti-Lamin B2 antibody [LN43] (ab8983)
      Immunohistochemistry (Frozen sections) - Anti-Lamin B2 antibody [LN43] (ab8983)

      IF staining of a 9 days old zebrafish embryo.  

    References

    This product has been referenced in:

    • Rao RA  et al. KMT1 family methyltransferases regulate heterochromatin-nuclear periphery tethering via histone and non-histone protein methylation. EMBO Rep 20:N/A (2019). Read more (PubMed: 30858340) »
    • Sen Gupta A  et al. Nucleolin modulates compartmentalization and dynamics of histone 2B-ECFP in the nucleolus. Nucleus 9:350-367 (2018). Read more (PubMed: 29943658) »
    See all 27 Publications for this product

    Publishing research using ab8983? Please let us know so that we can cite the reference in this datasheet.

    Customer reviews and Q&As

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    1-10 of 12 Abreviews or Q&A

    Immunocytochemistry/ Immunofluorescence abreview for Anti-Lamin B2 antibody [LN43]

     Good
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (Hela K)
    Permeabilization
    Yes - CSK buffer
    Specification
    Hela K
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: RT°C
    Fixative
    Paraformaldehyde
    Read More

    Abcam user community

    Verified customer

    Submitted Nov 09 2015

    Immunocytochemistry/ Immunofluorescence abreview for Anti-Lamin B2 [LN43] antibody

     Average
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 24°C
    Sample
    Mouse Cell (Neuronal progenitor cell line)
    Specification
    Neuronal progenitor cell line
    Permeabilization
    Yes - PBS 1% BSA 0,1% Saponine
    Fixative
    Paraformaldehyde
    Read More

    Abcam user community

    Verified customer

    Submitted Apr 08 2014

    Western blot abreview for Anti-Lamin B2 [LN43] antibody

     Good
    Abreviews
    abreview image
    Application
    Western blot
    Loading amount
    20 µg
    Gel Running Conditions
    Non-reduced Denaturing (precast 4-15%)
    Sample
    Human Cell lysate - whole cell (breast cell line)
    Specification
    breast cell line
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5%
    Read More

    Abcam user community

    Verified customer

    Submitted Mar 20 2014

    Question

    No signal in mouse olfactory tissue.

    Read More
    Answer

    Apologies for the misunderstanding but I thought you wanted a replacement instead of a refund. As requested I have credited the cost of ab8983. Your credit note number is ******.

    If you have any further questions on the credit note, please contact our accounts team at mailto:us.accounts@abcam.com and they will be happy to help.

    Please let me know if there is anything else I can help you with.

    Read More

    Question

    No signal in mouse olfactory tissue.

    Read More
    Answer

    Thank you for contacting Abcam yesterday.

    As we discussed I am sending you a vial of ab97513 as a replacement for ab8983. Your new order/reference number is ********** and you should have it tomorrow morning.

    If there is anything else I can help you with, please let me know.

    Read More

    Immunocytochemistry/ Immunofluorescence abreview for Anti-Lamin B2 antibody [LN43] - Nuclear Envelope Marker

     Below Average
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Xenopus laevis Cell (nuclei assembled in egg extract)
    Specification
    nuclei assembled in egg extract
    Fixative
    Paraformaldehyde
    Permeabilization
    No
    Blocking step
    BSA as blocking agent for 18 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 4°C
    Read More

    Abcam user community

    Verified customer

    Submitted Oct 28 2010

    Western blot abreview for Anti-Lamin B2 antibody [LN43] - Nuclear Envelope Marker

     Good
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Xenopus laevis Cell lysate - whole cell (meiotic egg extract)
    Loading amount
    25 µg
    Specification
    meiotic egg extract
    Gel Running Conditions
    Reduced Denaturing (15%)
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Read More

    Abcam user community

    Verified customer

    Submitted Oct 28 2010

    Question

    Our lab has recently purchased the Anti-Lamin B2 antibody (catalogue #: ab8983) and we would like to also purchase the immunizing peptide corresponding to this antibody. However, there is no immunizing peptide listed on the data sheet for the Anti-Lamin B2 antibody. I was wondering if you have an immunizing peptide to this antibody and if so what is its catalogue number? Thank you,

    Read More
    Answer

    Thank you for your enquiry. Unfortunately, this product was not made from an immunizing peptide, but from a detergent insoluble fraction of potoroo cell line PtK1. Therefore, the immunizing peptide does not exist for this product. I am sorry I can't be of more assistance in this occasion but if there is anything else that I can help you with, please do not hesitate to contact me.

    Read More

    Question

    I tested an antibody ab9883 Lamin B2 on rat tissue. 60ug of nuclear extraction prepared from rat brain was loaded on 15% of tris polyacrylamide gel. I used Precision Plus Protein Standards Dual color Marker, which recognizes protein bands between 250kDa and 10 kDa. Lamin B2 was diluted 1:100. A secondary antibody , alexa Fluor 680 goat anti mouse, was choosen to incubate a membrane 1 hr at room temperature with dilution 1:4000. Li-COR system showed that only one band with the size 50 kDa was detected. No other bands around it! So, does anyone have comments for that? Your product description says that a band should be detected approximately 67-65 kDa on mouse human hamster and Xenopus laevis.

    Read More
    Answer

    Thank you for your feeback on your experience of ab8983 in your rat brain samples. We have not tested the antibody in this species so it maybe that the protein is truncated or smaller in this species or tissue. Brain contains relatively few nuclei and is therefore not the preferable tissue for the isolation of lamins, I therefore think it is best to run a positive control along your samples, such as NCI-H125 nuclear cell lysate as used in the paper by Machiels BM et al.( Nuclear lamin expression in normal testis and testicular germ cell tumours of adolescents and adults. J Pathol 182:197-204 (1997)). I have included on the datasheet the protocols from that paper to help you, please click on the link below which will take you to the datasheet. I hope this information will help you, please let us know how you get on, we look forward to receiving your review,

    Read More

    Question

    BATCH NUMBER 125859 ORDER NUMBER 20004851 DESCRIPTION OF THE PROBLEM No signal or weak signal SAMPLE mouse fibroblast cell extract PRIMARY ANTIBODY 1:50 and 1:100 in blocking solution, incubate at 4C overnight, wash 3x 10 minutes times DETECTION METHOD ECLPlus ANTIBODY STORAGE CONDITIONS -20 C SAMPLE PREPARATION TGS buffer PMSF, Leupeptin and Pepstatin, Heating to 100C for 10 minutes AMOUNT OF PROTEIN LOADED 15-30 ul/lane ELECTROPHORESIS/GEL CONDITIONS 10, 12, 15% gels TRANSFER AND BLOCKING CONDITIONS 5%milk or 5% BSA for 1 hour at room T SECONDARY ANTIBODY [competitor] sheep anti mouse Ig, 1:2500 to 1:5000, room T for 1 hour, wash 3x 10 minutes times HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 10 HAVE YOU RUN A "NO PRIMARY" CONTROL? Yes DO YOU OBTAIN THE SAME RESULTS EVERY TIME? Yes WHAT STEPS HAVE YOU ALTERED? Blocking solution, Amount of protein loaded, different percentages of gels, Membrane, primery Ab concentration, second Ab concentration ADDITIONAL NOTES We have bought 2 aliquots of this Ab from your campany so far but none of them works on western blot. I striped my membranes and reprobe for other proteins and they work perfactly. ( I used the same 2ed Ab).

    Read More
    Answer

    Thank you for your enquiry and I'm sorry to hear that you are experiencing difficulty with ab8983. At this point I would like to make the following suggestions. Try using nuclear extract rather than whole cell extract, and with the nuclear extract incubate overnight at 4C with the primary diluted to 1:50. Also, I see that you mentioned loading 15-30 ul/lane - do you mean 15-30 ug protein per lane? Please make sure that you are loading at least 30 ug protein. If these suggestions do not help and you have purchased the antibody in the past 90 days I would be happy to issue a refund or credit note to you. Have a nice weekend!

    Read More

    1-10 of 12 Abreviews or Q&A

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