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遺伝子ノックアウト細胞株 検証済

Anti-Lamin B1 抗体 [119D5-F1] - Nuclear Envelope Marker (ab8982)

Reviews (6)Q&A (9)References (42)
Immunocytochemistry/ Immunofluorescence - Anti-Lamin B1 antibody [119D5-F1] - Nuclear Envelope Marker (ab8982)
  • Immunohistochemistry (Frozen sections) - Anti-Lamin B1 antibody [119D5-F1] - Nuclear Envelope Marker (ab8982)
  • Immunohistochemistry (Frozen sections) - Anti-Lamin B1 antibody [119D5-F1] - Nuclear Envelope Marker (ab8982)
  • Immunocytochemistry/ Immunofluorescence - Anti-Lamin B1 antibody [119D5-F1] - Nuclear Envelope Marker (ab8982)
  • Flow Cytometry - Anti-Lamin B1 antibody [119D5-F1] - Nuclear Envelope Marker (ab8982)
  • Immunohistochemistry (Frozen sections) - Anti-Lamin B1 antibody [119D5-F1] - Nuclear Envelope Marker (ab8982)
  • Immunohistochemistry (Frozen sections) - Anti-Lamin B1 antibody [119D5-F1] - Nuclear Envelope Marker (ab8982)

Key features and details

  • Mouse monoclonal [119D5-F1] to Lamin B1 - Nuclear Envelope Marker
  • Suitable for: Flow Cyt, ICC/IF, IHC-Fr
  • Knockout validated
  • Reacts with: Mouse, Human, Pig
  • Isotype: IgG1

製品の概要

  • 製品名

    Anti-Lamin B1 antibody [119D5-F1] - Nuclear Envelope Marker
    Lamin B1 一次抗体 製品一覧

  • 製品の詳細

    Mouse monoclonal [119D5-F1] to Lamin B1 - Nuclear Envelope Marker

  • 由来種

    Mouse

  • 特異性

    Reacts with an epitope located C-terminal of residue 231 in lamin B1. Reacts against lamin B1, does not cross react with lamin B2. Lamins do not appear to be universally distributed among different cell and tissue types. ab8982 has been shown to react with HeLa and 3T3 cells in immunocytochemistry. Other cell/tissue types have not been tested.

    High levels of background are often observed when staining mouse tissue with a mouse antibody. We recommend ab16048 as an alternative for mouse staining.

  • アプリケーション

    適用あり: Flow Cyt, ICC/IF, IHC-Frmore details

  • 種交差性

    交差種: Mouse, Human, Pig

  • 免疫原

    Full length native protein (purified) corresponding to Rat Lamin B1 (C terminal). Immunogen is purified rat liver lamins.

  • エピトープ

    C-terminal to residue 231.

  • ポジティブ･コントロール

    • ICC/IF: HAP1, HeLa and 3T3 cells. WB: Human mammary cell lysate. IHC-Fr: Human kidney and colon tissue; MCF7 cell culture; Swine liver tissue. Flow Cyt: HeLa cells

  • 特記事項

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. 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, along with publications, customer reviews and Q&As

製品の特性

アプリケーション

The Abpromise guarantee

Abpromise保証は、 次のテスト済みアプリケーションにおけるab8982の使用に適用されます

アプリケーションノートには、推奨の開始希釈率がありますが、適切な希釈率につきましてはご検討ください。

アプリケーション Abreviews 特記事項
Flow Cyt
1/100 - 1/200.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
ICC/IF
Use a concentration of 0.5 µg/ml.
IHC-Fr
Use at an assay dependent concentration.
特記事項
Flow Cyt
1/100 - 1/200.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
ICC/IF
Use a concentration of 0.5 µg/ml.
IHC-Fr
Use at an assay dependent concentration.

ターゲット情報

  • 機能

    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.

  • 関連疾患

    Defects in LMNB1 are the cause of leukodystrophy demyelinating autosomal dominant adult-onset (ADLD) [MIM:169500]. ADLD is a slowly progressive and fatal demyelinating leukodystrophy, presenting in the fourth or fifth decade of life. Clinically characterized by early autonomic abnormalities, pyramidal and cerebellar dysfunction, and symmetric demyelination of the CNS. It differs from multiple sclerosis and other demyelinating disorders in that neuropathology shows preservation of oligodendroglia in the presence of subtotal demyelination and lack of astrogliosis.

  • 配列類似性

    Belongs to the intermediate filament family.

  • 翻訳後修飾

    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.

  • 細胞内局在

    Nucleus inner membrane.
  • Information by UniProt

  • 参照データベース

    • 別名

      • ADLD antibody
      • lamin B1 antibody
      • Lamin-B1 antibody
      • LMN antibody
      • LMN2 antibody
      • LMNB antibody
      • Lmnb1 antibody
      • LMNB1_HUMAN antibody
      • MGC111419 antibody
      • OTTHUMP00000159218 antibody
      see all

    画像

    • Immunocytochemistry/ Immunofluorescence - Anti-Lamin B1 antibody [119D5-F1] - Nuclear Envelope Marker (ab8982)
      Immunocytochemistry/ Immunofluorescence - Anti-Lamin B1 antibody [119D5-F1] - Nuclear Envelope Marker (ab8982)

      ab8982 staining Lamin B1 in wild-type HAP1 cells (top panel) and LMNB1 knockout HAP1 cells (bottom panel). The cells were fixed with 100% methanol (5min), permeabilized with 0.1% Triton X-100 for 5 minutes 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 ab8982 at 0.5μg/ml and ab202272 at 1/250 dilution (shown in pseudocolour red) overnight at +4°C, followed by a further incubation at room temperature for 1h with a goat secondary antibody to Mouse IgG (Alexa Fluor® 488) (ab150117) at 2 μg/ml (shown in green). Nuclear DNA was labelled in blue with DAPI.

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

    • Immunohistochemistry (Frozen sections) - Anti-Lamin B1 antibody [119D5-F1] - Nuclear Envelope Marker (ab8982)
      Immunohistochemistry (Frozen sections) - Anti-Lamin B1 antibody [119D5-F1] - Nuclear Envelope Marker (ab8982)

      Immunohistochemistry on frozen sections of human kidney showing nuclear lamina staining in epithelial and connective tissue cells.

    • Immunohistochemistry (Frozen sections) - Anti-Lamin B1 antibody [119D5-F1] - Nuclear Envelope Marker (ab8982)
      Immunohistochemistry (Frozen sections) - Anti-Lamin B1 antibody [119D5-F1] - Nuclear Envelope Marker (ab8982)

      Immunohistochemistry of MCF-7 cell culture showing nuclear lamina staining.

    • Immunocytochemistry/ Immunofluorescence - Anti-Lamin B1 antibody [119D5-F1] - Nuclear Envelope Marker (ab8982)
      Immunocytochemistry/ Immunofluorescence - Anti-Lamin B1 antibody [119D5-F1] - Nuclear Envelope Marker (ab8982)This image is courtesy of Marilena Ciciarello & Patrizia Lavia, University La Sapienza

      Methanol fixed HeLa and 3T3 cells (ab7179) were stained with ab8982 (1/50 and 1/20 respectively). The cells were fixed in 100% methanol for 6 minutes at -20°C. ab8982 clearly stains the nuclear envelope with very little background staining. 

      A: HeLa cells + ab8982 (1/50) (green)
      B: HeLa cells counterstained with DAPI (blue)
      C. 3T3 cells + ab8982 (1/20) (green)
      D. 3T3 cells counterstained with DAPI (blue)

    • Flow Cytometry - Anti-Lamin B1 antibody [119D5-F1] - Nuclear Envelope Marker (ab8982)
      Flow Cytometry - Anti-Lamin B1 antibody [119D5-F1] - Nuclear Envelope Marker (ab8982)

      Overlay histogram showing HeLa cells stained with ab8982 (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 (ab8982, 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 B1 antibody [119D5-F1] - Nuclear Envelope Marker (ab8982)
      Immunohistochemistry (Frozen sections) - Anti-Lamin B1 antibody [119D5-F1] - Nuclear Envelope Marker (ab8982)

      Immunohistochemistry on frozen sections of swine liver showing nuclear lamina staining in hepatocytes.

    • Immunohistochemistry (Frozen sections) - Anti-Lamin B1 antibody [119D5-F1] - Nuclear Envelope Marker (ab8982)
      Immunohistochemistry (Frozen sections) - Anti-Lamin B1 antibody [119D5-F1] - Nuclear Envelope Marker (ab8982)

      Immunohistochemistry on frozen sections of human colon showing nuclear lamina staining in epithelial and connective tissue cells.

    プロトコール

    To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

    Click here to view the general protocols

    参考文献 (42)

    ab8982 を使用した論文を発表された方は、こちらまでお知らせください。データシートに掲載させていただきます。

    ab8982 は 42 報の論文で使用されています。

    • Palmieri M  et al. PI3Kα Translocation Mediates Nuclear PtdIns(3,4,5)P3 Effector Signaling in Colorectal Cancer. Mol Cell Proteomics 22:100529 (2023). PubMed: 36931626
    • Thomas MA  et al. An unbiased proteomic analysis of PAD4 in human monocytes: novel substrates, binding partners and subcellular localizations. Philos Trans R Soc Lond B Biol Sci 378:20220477 (2023). PubMed: 37778379
    • Meharena HS  et al. Down-syndrome-induced senescence disrupts the nuclear architecture of neural progenitors. Cell Stem Cell 29:116-130.e7 (2022). PubMed: 34995493
    • Muralidharan H  et al. KIFC1 Regulates the Trajectory of Neuronal Migration. J Neurosci 42:2149-2165 (2022). PubMed: 35046122
    • Wang YF  et al. Seawater immersion exacerbates the pathological changes caused by incisive corneal injury in rabbit eyes. Ann Transl Med 10:589 (2022). PubMed: 35722388
    View all Publications for this product

    レビューと Q&A

    Show All レビュー Q&A
    レビューを送る 質問を送る

    1-10 of 15 Abreviews or Q&A

    Western blot abreview for Anti-Lamin B1 antibody [119D5-F1] - Nuclear Envelope Marker

     Average
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Boa constrictor Cell lysate - whole cell (kidney and brain cell types)
    Gel Running Conditions
    Reduced Denaturing (12%)
    Loading amount
    40 µg
    Specification
    kidney and brain cell types
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    投稿 Dec 17 2019

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Lamin B1 antibody [119D5-F1] - Nuclear Envelope Marker

     Good
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Mouse Tissue sections (Brain)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Tri-Sodium Citrate
    Permeabilization
    Yes - with blocking
    Specification
    Brain
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Fixative
    Paraformaldehyde
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Juliana Schneider

    Verified customer

    投稿 Mar 28 2017

    Western blot abreview for Anti-Lamin B1 antibody [119D5-F1] - Nuclear Envelope Marker

     Poor
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Mouse Cell lysate - nuclear (Muscle)
    Gel Running Conditions
    Reduced Denaturing
    Loading amount
    40 µg
    Specification
    Muscle
    Blocking step
    Seablock as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 27°C
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    投稿 Nov 17 2016

    Western blot abreview for Anti-Lamin B1 antibody [119D5-F1] - Nuclear Envelope Marker

     Good
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Tissue lysate - whole (Brain)
    Gel Running Conditions
    Non-reduced Denaturing (8%)
    Loading amount
    15 µg
    Specification
    Brain
    Blocking step
    5% BSA + 5% Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    投稿 Oct 27 2014

    Western blot abreview for Anti-Lamin B1 antibody [119D5-F1] - Nuclear Envelope Marker

     Good
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (Human mammary cell)
    Gel Running Conditions
    Reduced Denaturing (8%)
    Loading amount
    50000 cells
    Specification
    Human mammary cell
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    投稿 Feb 22 2012

    Question

    Dear Sir/Madam
    Thank you for your prompt response to Katarzyna Bielska concerning problems with Anti-Lamin B1 antibody (cat No ab8982). Of course, we prefer a new vial of your product as a free charge replacement. Because I’ve ordered your product, please send a new vial to the address below.

    Best regards

    Read More
    Answer

    Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products.

    I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement with the order number 1136297.

    To check the status of the order please contact our Customer Service team and reference this number.

    Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know.

    I wish you the best of luck with your research.

    Read More

    Question

    I probably will ask for a new vial. I go to the holiday next week, so my colleague, will contact with you.
    Thank you for your help.
    Best regards

    Read More
    Answer

    Have a lovely holiday and please ask your colleague to confirm if you wish to get a replacement and when.

    I look forward to hearing from you soon.

    Read More

    Question

    A) Image: Both images labelled as PPAR. I am a bit confused. Could you attach an image for Lamin B1 which has the MW markers labelled properly?

    This is the problem, that I don't see lamin B1 - my control. I want to analyze levels of PPAR. I don't have problems with PPAR antibody but lamin B.

    B) Lysis buffer: Has the customer used RIPA buffer which contains not one - but a mixture of different proteinase inhibitors. It could be that the lower band (on the top image at around 55 kDa) represent some partially degraded protein.
    I have checked differnt coctails of inhibitors and PMSF give the best results.
    C) Dilution: Has the customer checked this antibody at a dilution of 1/100 to see if the intensity of the signal changes?
    I haven't. I don't think it will change something, if I don't see bands at a dilution 1/200.
    Best regards

    Read More
    Answer

    Thank you for getting back to me and for providing some further details.

    I would like to reassure you that our Abpromise applies to your complaint since you purchased this product within the guarantee period. This means that in the event that a product is not functioning in the applications/species cited on the product data sheet (and the problem has been reported within 6 months of purchase) we will happily offer a credit note/refund to the value of the product purchased.

    I could offer you either a new vial as a free charge replacement or a credit note which you can use in the future. Please do let me know how you wish to proceed with your enquiry.

    I look forward to hearing from you soon.

    Read More

    Question

    1) Abcam product code ab 8982

    2) Abcam order reference number or product batch number lot GR43517-7

    3) Description of the problem
    I isolate nuclei and prepare nuclear protein extracts from rabbit liver. I have planned to use lamina B as my control. However, I don’t get signals from lamina B.

    4) Sample preparation:
    Type of sample (whole cell lysates, fraction, recombinant protein…) nuclear protein extracts
    Lysis buffer

    124 mM Tris-HCl, 20% glycerol, 4% SDS, 10% b-mercaptoethanol, 0,005% bromophenol blue, pH 6.8

    Protease inhibitors: PMSF
    Phosphatase inhibitors
    Reducing agent
    Boiling for ≥5 min? yes/no
    Protein loaded ug/lane or cells/lane 30 mg/lane
    Positive control
    Negative control

    5) Percentage of gel 4% and 10%
    Type of membrane nitrocellulose
    Protein transfer verified Ponceau
    Blocking agent and concentration 5% milk in PBST buffer
    Blocking time 2 h
    Blocking temperature RT – room temperature

    6) Primary antibody (If more than one was used, describe in “additional notes”) : anti lamin B
    Concentration or dilution 1:200
    Diluent buffer 5% milk in PBST buffer
    Incubation time 14 h
    Incubation temperature: 4°C

    7) Secondary antibody: HAMPO
    Species: horse
    Reacts against: mouse
    Concentration or dilution 1:10 000
    Diluent buffer 5% milk in PBST buffer
    Incubation time 2h
    Incubation temperature: RT
    Fluorochrome or enzyme conjugate: HRP

    8) Washing after primary and secondary antibodies:
    Buffer PBST
    Number of washes 3 for 10 minutes

    9) Detection method
    The fluorescence signal was recorded with the Fuji Film FLA-3000 Fluorescent Image Analyzer (Raytest Isotopenmebgera¨te GmbH).
    I also used Biomax Light Film (Kodak).

    10) How many times have you run this staining? 2
    Do you obtain the same results every time? yes
    What steps have you altered to try and optimize the use of this antibody? I only change the detection method.


    11) “additional notes”
    I also used PPAR antibody (primary antibody), but I didn’t have problems with detection.

    Read More
    Answer

    Thank you for your enquiry regarding ab8982 and for taking the time to provide some useful details of the experiments. I am very sorry to hear that your customer is having problems with this antibody.

    A) Image: Both images labelled as PPAR. I am a bit confused. Could you attach an image for Lamin B1 which has the MW markers labelled properly?

    B) Lysis buffer: Has the customer used RIPA buffer which contains not one - but a mixture of different proteinase inhibitors. It could be that the lower band (on the top image at around 55 kDa) represent some partially degraded protein.

    C) Dilution: Has the customer checked this antibody at a dilution of 1/100 to see if the intensity of the signal changes?

    I look forward to hearing from you soon.

    Read More

    Question

    Dear Sir/Madam
    I am working with antibody ab 8982 (lot GR43517-7). I isolate nuclei and prepare nuclear protein extracts from rabbit liver. I have planned to use lamina B as my control. However, I don’t get signals from lamina B, but I can see other nuclear proteins. I store the antibody at +4°C and I have worked with the dilution 1:200. I have checked secondary antibody (they work). Do you have any information that somebody had problems with Anti-Lamin B1 antibody (lot GR43517-7)?
    Best regards

    Read More
    Answer

    I am sorry to hear that you have been experiencing problems using this product in the application that you wish.

    All our customer feedback, including complaints are monitored weekly by our in house technical support team. If a product is at fault the technical support team will consider removing the product from our catalogue in order to avoid future customer inconvenience. We have no complaint about this batch (1216925) at all.

    As our Abpromise indicates, in the event that a product is not functioning in the applications/species cited on the product data sheet (and the problem has been reported within 6 months of purchase) we will happily offer a credit note/refund to the value of the product purchased

    Could you please provide some further details of the protocol used and complete the following form (attached as a word document). It would be much appreciated if you could attach an image to the response.

    Thank you for your understanding and co-operation in this matter. I look forward to hearing from you soon and resolving this issue as soon as possible.

    Read More

    1-10 of 15 Abreviews or Q&A

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