Anti-LAP2 alpha 抗体 (ab5162)

Reviews (1)Q&A (5)References (16)
Immunocytochemistry/ Immunofluorescence - Anti-LAP2 alpha antibody (ab5162)
  • Western blot - Anti-LAP2 alpha antibody (ab5162)

Key features and details

  • Rabbit polyclonal to LAP2 alpha
  • Suitable for: ICC/IF, WB
  • Reacts with: Human
  • Isotype: IgG

製品の概要

  • 製品名

    Anti-LAP2 alpha antibody
    LAP2 alpha 一次抗体 製品一覧

  • 製品の詳細

    Rabbit polyclonal to LAP2 alpha

  • 由来種

    Rabbit

  • アプリケーション

    適用あり: ICC/IF, WBmore details

  • 種交差性

    交差種: Human

  • 免疫原

    Recombinant fragment (His-tag) corresponding to Human LAP2 alpha aa 188-693.

  • 特記事項


    Lamins are type V intermediate filament proteins and are grouped into constitutively expressed B-type lamins and developmentally regulated A- type lamins. Lamin-binding proteins in the nuclear lamina and the nuclear interior include several protein families and/or types of proteins in higher eu karyotes such as the inner nuclear membrane proteins, lamin B receptor, emerin, and MANI, three isoforms of lamina-associated polypeptide 1 (LAP 1), and several isoforms of LAP 2. Up to six LAP 2 isoforms derive from a single gene by alternative splicing in mammals and various isoforms have been described in Xenopus. The best characterized LAP2 isoforms are the inner nuclear membrane protein LAP 2 beta and the nucleoplasmic protein LAP 2 alpha, which are identical in their N-terminal 187-amino acid constant region but differ in their C termini. While LAP 2 beta binds to B-type lamins at the nuclear periphery and was suggested to regulate nuclear lamina growth , LAP 2 alpha specifically interacts with A-type lamins within the nuclear interior as part of a detergent/salt-resistant nucleoskeletal structure.

    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

  • 製品の状態

    Liquid

  • 保存方法

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.

  • バッファー

    Preservative: 0.1% Sodium azide
    Constituents: PBS, Whole serum
  • Concentration information loading...

  • 精製度

    Whole antiserum

  • 一次抗体 備考

    Lamins are type V intermediate filament proteins and are grouped into constitutively expressed B-type lamins and developmentally regulated A- type lamins. Lamin-binding proteins in the nuclear lamina and the nuclear interior include several protein families and/or types of proteins in higher eu karyotes such as the inner nuclear membrane proteins, lamin B receptor, emerin, and MANI, three isoforms of lamina-associated polypeptide 1 (LAP 1), and several isoforms of LAP 2. Up to six LAP 2 isoforms derive from a single gene by alternative splicing in mammals and various isoforms have been described in Xenopus. The best characterized LAP2 isoforms are the inner nuclear membrane protein LAP 2 beta and the nucleoplasmic protein LAP 2 alpha, which are identical in their N-terminal 187-amino acid constant region but differ in their C termini. While LAP 2 beta binds to B-type lamins at the nuclear periphery and was suggested to regulate nuclear lamina growth , LAP 2 alpha specifically interacts with A-type lamins within the nuclear interior as part of a detergent/salt-resistant nucleoskeletal structure.

  • ポリ/モノ

    ポリクローナル

  • アイソタイプ

    IgG

  • 研究分野

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ab5162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
ICC/IF
Use at an assay dependent concentration.
WB (1)
1/2500. Detects a band of approximately 83 kDa.
特記事項
ICC/IF
Use at an assay dependent concentration.
WB
1/2500. Detects a band of approximately 83 kDa.

ターゲット情報

  • 関連性

    Lamins are type V intermediate filament proteins and are grouped into constitutively expressed B-type lamins and developmentally regulated A-type lamins. Lamin-binding proteins in the nuclear lamina and the nuclear interior include several protein families and/or types of proteins in higher eukaryotes such as the inner nuclear membrane proteins, lamin B receptor, emerin, MANI, three isoforms of lamina-associated polypeptide 1 (LAP 1), and several isoforms of LAP 2. Up to six LAP 2 isoforms derive from a single gene by alternative splicing in mammals and various isoforms have been described in Xenopus. The best characterized LAP2 isoforms are the inner nuclear membrane protein LAP 2 beta and the nucleoplasmic protein LAP 2 alpha, which are identical in their N-terminal 187-amino acid constant region but differ in their C termini. LAP 2 alpha specifically interacts with A-type lamins within the nuclear interior as part of a detergent- and salt-resistant nucleoskeletal structure.

  • 細胞内局在

    Nuclear

  • 参照データベース

    • 別名

      • Isoforms beta/gamma antibody
      • Lamina associated polypeptide 2 antibody
      • LAP2 antibody
      • Thymopoietin antibody
      • Thymopoietin isoform alpha antibody
      • TMPO antibody
      • TP alpha antibody
      see all

    画像

    • Immunocytochemistry/ Immunofluorescence - Anti-LAP2 alpha antibody (ab5162)
      Immunocytochemistry/ Immunofluorescence - Anti-LAP2 alpha antibody (ab5162)

      ab5162 at a dilution of 1/250 staining LAP 2 alpha in HeLa cells by immunofluorescence.

      A = ab5162, 1/250

      B = DNA

      C = A and B merged

    • Western blot - Anti-LAP2 alpha antibody (ab5162)
      Western blot - Anti-LAP2 alpha antibody (ab5162)
      ab5162 at dilutions of 1/100 and 1/500 staining approximately 83 kDa LAP 2 alpha in HeLa cell extracts by Western blot (ECL). ab5162 at dilutions of 1/100 and 1/500 staining approximately 83 kDa LAP 2 alpha in HeLa cell extracts by Western blot (ECL).

    参考文献 (16)

    ab5162 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。

    ab5162 は 16 報の論文で使用されています。

    • Chojnowski A  et al. Heterochromatin loss as a determinant of progerin-induced DNA damage in Hutchinson-Gilford Progeria. Aging Cell 19:e13108 (2020). PubMed: 32087607
    • Nastaly P  et al. Role of the nuclear membrane protein Emerin in front-rear polarity of the nucleus. Nat Commun 11:2122 (2020). PubMed: 32358486
    • Mirza AN  et al. LAP2 Proteins Chaperone GLI1 Movement between the Lamina and Chromatin to Regulate Transcription. Cell 176:198-212.e15 (2019). PubMed: 30503211
    • Qin Z  et al. Long noncoding RNA TMPO-AS1 promotes progression of non-small cell lung cancer through regulating its natural antisense transcript TMPO. Biochem Biophys Res Commun 516:486-493 (2019). PubMed: 31230752
    • Wada E  et al. Deficiency of emerin contributes differently to the pathogenesis of skeletal and cardiac muscles in LmnaH222P/H222P mutant mice. PLoS One 14:e0221512 (2019). PubMed: 31430335
    View all Publications for this product

    レビューと Q&A

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    1-6 of 6 Abreviews or Q&A

    Western blot abreview for Anti-LAP2 alpha antibody

     Good
    Abreviews
    abreview image
    Application
    Western blot
    Loading amount
    20 µg
    Gel Running Conditions
    Reduced Denaturing
    Sample
    Human Cell lysate - whole cell (U2OS)
    Specification
    U2OS
    Blocking step
    Milk as blocking agent for 3 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
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    Abcam user community

    Verified customer

    投稿 Oct 22 2014

    Question

    Product code: 5162
    Lot number: GR17174-2

    Inquiry: Good afternoon, We are routinely using the LAP2alpha antibody #ab5162 for WB and more recently for CoIP. Lot number as indicated on our order GR17174 (N/A) For the latest application, we need to know the concentration of the antibody which unfortunately appears as 'Not available' on the data sheet. Please could you let us know what the concentration is in this lot and also confirm us that concentration will be available in subsequent orders. Sincerely yours,

    Read More
    Answer

    Thank you for your enquiry.

    I can confirm that ab5162(Anti-LAP2 alpha antibody) is sold as whole antiserum. Unpurified antibodies, such as those sold as whole antiserum, ascites or tissue culture supernatant will not have a concentration stated on the datasheet.

    Antibody concentration is usually determined by protein assay, and serum will contain a lot of other proteins, which means the antibody quantification would not be accurate.

    I can confirm that for whole antiserum, concentration of antibody is known to vary between 1 - 5 mg/ml.

    I am sorry we are not able to provide an exact concentration on this occasion, but hope this information will be helpful to you. If you have any further questions, please do not hesitate to contact us.

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    Question

    We will accept the offer to try another vial from the same batch
    Best regards

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    Answer

    Thank you for kindly confirming these details. I am sorry this vial of antibody has not worked for this customer.

    As requested, we are pleased to arrange a free of charge replacement. I can confirm that 1 vial of ab5162 has been added to your order number #####. This has Abcam order reference ######.

    I would like to reassure you that this free of charge replacement vial is also covered by our Abpromise guarantee. Should the customer still be experiencing difficulties with the new vial, or if you have any further questions, please do not hesitate to let me know.

    Thank you for your help and cooperation with this case. Please do not hesitate to contact me if you need anything further.

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    Question

    The customer would like to try different batch

    Please advise
    Best regards

    Read More
    Answer

    Thank you for your reply.

    I am sorry to confirm that we don't have any different lots in stock at the moment. I would like to reassure you that we monitor feedback closely on a weekly basis and we are not currently concerned about the general quality of this antibody or this batch. Regrettably, I can suggest you have received a bad vial on this occasion.

    Therefore, we would be pleased to provide a replacement from the same lot, which will still be covered by our guarantee. Alternatively, I fully understand the customers concerns and you prefer a credit note in this case, I will be pleased to arrange this for you.

    I hope this will be helpful. I look forward to hearing from you with details of how you would like to proceed.

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    Question

    Dear Kate

    Our response was probably delayed

    Here are the answers:

    customer:

    The incubation time was1h at room temperature

    In the same experiment, under the same conditions I used laminA antibody ( the same order number) and got wonderful results.

    I need Lap2Alpha antibody, and I will be very happy to get a sample of it from other bench.

    Let me know if you need anything else

    Regards

    Read More
    Answer

    Thank you for your message and for providing this further information.

    I am sorry to hear the suggestions made have not improved the results on this occasion. I appreciate the time you have spent on these experiments and would be pleased to arrange a free of charge replacement or credit note in compensation.

    I am sorry that because of the number of products we have on the catalog, it is not possible for us to store sample vials to send to customers. I can reassure you that any replacement vial would also be covered by the 6 month guarantee.

    I look forward to hearing from you with details of how you would like to proceed.

    Read More

    Question

    Product code: 5162
    Lot number: GR17174-4
    Inquiry: Please read below the details of customer's complaint:
    Antibody code: ab5162
    Batch number: Lot GR 17174-4
    General Information:
    Antibody storage conditions (temperature/reconstitution etc) -20C

    Description of the problem: strong background, also on the cells with Lap2Alpha depletion (Lap2Alpha-/-)
    Sample (Species/Tissue/Cell Type/Cell Line etc.) MEF cells

    Fixation of sample (Ethanol/Methanol/Acetone/Paraformaldehyde/Other/Duration etc.) Paraformaldehyde

    Permeabilization step 3 min with 0.5 %Triton

    Blocking conditions (Buffer/time period, Blocking agent etc.) 5%BSA for 20min

    Secondary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step) Invitrogen Alexa Fluor 488 goat antirabbit (H+L), 1:1400, incubation 60min, 3 times washing*5min

    Detection method Confocal microscope

    Positive and negative controls used (please specify) cells with Lap2Alpha-/- and Lap2Alpha+/+ were used. In the cells with Lap2Alpha depletion there were no strong staining in the nucleus, but t non specific signal was detected from the cytoplasm. Secondary antibody was used in the same experiment for LaminA-/- and LaminA +/+ and it gave perfect results.

    Optimization attempts (problem solving): different delitopns of Lap2 alpha antibody (did not help)

    How many times have you tried the ICC? 2times, ech time the same result, perfect for LaminA antibody and with high background for Lap2lpha antibody.

    Have you run a "No Primary" control? not

    Do you obtain the same results every time? yes

    What steps have you altered? Additional Notes We bought 2 antibody and run the experiment simultaneously with both of them LaminA antibody gave fantastic results, while Lap2Alpha was not specific, even in the cells with Lap2Alpha depletion.

    Thanks in advance for your advise.
    Regards,

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    Answer

    Thank you for taking the time to complete our questionnaire and contact us. I am sorry to hear you have had difficulty obtaining satisfactory results from this antibody.

    The details you have kindly provided will enable us to investigate this case for you and this is also helpful in our records for monitoring of quality.

    I would like to reassure you that ab5162 is tested and covered by our 6 month guarantee for use in ICC-IF and mouse samples. In the event that a product is not functioning in the applications cited on the product data sheet, we will be pleased to provide a credit note or free of charge replacement.

    Although the other antibody has worked well using this procedure, individual antibodies will often require optimization. I would like to investigate this further and hopefully offer some suggestions to help optimise the results from ab5162. I would also appreciate if you can confirm some details of the procedure:

    1. Please provide the order number and date of purchase. Was this Po 88783 ordered 12th Feb?

    2. I would appreciate if you are able to provide further details of the primary antibody. What dilution was used, has this been optimized? How long was it incubated for and at what temperature?

    3. I would appreciate if you are able to provide an image which would help us to assess the results.

    Thank you for your cooperation. I look forward to hearing from you with the further requested details.

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