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Anti-Lysosomal acid lipase/LAL 抗体 [9G7F12] (ab36597)

Reviews (1)Q&A (3)References (3)
Western blot - Anti-Lysosomal acid lipase/LAL antibody [9G7F12] (ab36597)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Lysosomal acid lipase/LAL antibody [9G7F12] (ab36597)
  • Flow Cytometry - Anti-Lysosomal acid lipase/LAL antibody [9G7F12] (ab36597)

Key features and details

  • Mouse monoclonal [9G7F12] to Lysosomal acid lipase/LAL
  • Suitable for: Flow Cyt, WB, IHC-P
  • Reacts with: Human, Recombinant fragment
  • Isotype: IgG2a

製品の概要

  • 製品名

    Anti-Lysosomal acid lipase/LAL antibody [9G7F12]
    Lysosomal acid lipase/LAL 一次抗体 製品一覧

  • 製品の詳細

    Mouse monoclonal [9G7F12] to Lysosomal acid lipase/LAL

  • 由来種

    Mouse

  • アプリケーション

    適用あり: Flow Cyt, WB, IHC-Pmore details

  • 種交差性

    交差種: Human, Recombinant fragment

  • 免疫原

    Recombinant fragment corresponding to Lysosomal acid lipase/LAL.

  • 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ab36597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
Use 0.5µg for 106 cells.

ab170191 - Mouse monoclonal IgG2a, is suitable for use as an isotype control with this antibody.
WB (1)
1/500 - 1/2000. Detects a band of approximately 45 kDa (predicted molecular weight: 46 kDa).
IHC-P
Use a concentration of 2 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
特記事項
Flow Cyt
Use 0.5µg for 106 cells.

ab170191 - Mouse monoclonal IgG2a, is suitable for use as an isotype control with this antibody.
WB
1/500 - 1/2000. Detects a band of approximately 45 kDa (predicted molecular weight: 46 kDa).
IHC-P
Use a concentration of 2 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

ターゲット情報

  • 機能

    Crucial for the intracellular hydrolysis of cholesteryl esters and triglycerides that have been internalized via receptor-mediated endocytosis of lipoprotein particles. Important in mediating the effect of LDL (low density lipoprotein) uptake on suppression of hydroxymethylglutaryl-CoA reductase and activation of endogenous cellular cholesteryl ester formation.

  • 関連疾患

    Defects in LIPA are the cause of Wolman disease (WOD) [MIM:278000]. WOD is a severe manifestation of LIPA deficiency, leading to the accumulation of cholesteryl esters and triglycerides in most tissues of the body. WOD occurs in infancy and is nearly always fatal before the age of 1 year.
    Defects in LIPA are the cause of cholesteryl ester storage disease (CESD) [MIM:278000]. CESD is a mild manifestation of LIPA deficiency, leading to the accumulation of cholesteryl esters and triglycerides in most tissues of the body. It is characterized by late-onset.

  • 配列類似性

    Belongs to the AB hydrolase superfamily. Lipase family.

  • 細胞内局在

    Lysosome.
  • Information by UniProt

  • 参照データベース

    • 別名

      • Acid cholesteryl ester hydrolase antibody
      • CESD antibody
      • cholesterol ester hydrolase antibody
      • cholesterol ester storage disease antibody
      • Cholesteryl esterase antibody
      • Hydrolase deficiency antibody
      • LAL antibody
      • LAL deficiency cholesterol ester antibody
      • LICH_HUMAN antibody
      • lipA antibody
      • LIPA deficiency antibody
      • Lipase A antibody
      • lipase A, lysosomal acid, cholesterol esterase antibody
      • lysosomal acid lipase antibody
      • lysosomal acid lipase deficiency antibody
      • Lysosomal acid lipase/cholesteryl ester hydrolase antibody
      • Sterol esterase antibody
      see all

    画像

    • Western blot - Anti-Lysosomal acid lipase/LAL antibody [9G7F12] (ab36597)
      Western blot - Anti-Lysosomal acid lipase/LAL antibody [9G7F12] (ab36597)
      All lanes : Anti-Lysosomal acid lipase/LAL antibody [9G7F12] (ab36597)

      Lane 1 : Molecular weight marker
      Lane 2 : Truncated LAL recombinant protein

      Predicted band size: 46 kDa
      Observed band size: ~45 kDa why is the actual band size different from the predicted?

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Lysosomal acid lipase/LAL antibody [9G7F12] (ab36597)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Lysosomal acid lipase/LAL antibody [9G7F12] (ab36597)

      ab36597 (2 µg/ml) staining Lysosomal acid lipase/LAL in human lung using an automated system (DAKO Autostainer Plus). Using this protocol there is lysosomal staining in the bronchiolar epithelium and resident macrophages .
      Sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffer citrate pH 6.0 in a DAKO PT link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 mins. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 min and detected with Dako envision flex amplification kit for 30 minutes. Colorimetric detection was completed with Diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that, for manual staining, optimization of primary antibody concentration and incubation time is recommended. Signal amplification may be required.

    • Flow Cytometry - Anti-Lysosomal acid lipase/LAL antibody [9G7F12] (ab36597)
      Flow Cytometry - Anti-Lysosomal acid lipase/LAL antibody [9G7F12] (ab36597)
      Overlay histogram showing HepG2 cells stained with ab36597 (red line). The cells were fixed with 80% 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 / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab36597, 0.5µg/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 IgG2a [ICIGG2A] (ab91361, 1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.

    参考文献 (3)

    ab36597 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。

    ab36597 は 3 報の論文で使用されています。

    • Cui W  et al. Lipophagy-derived fatty acids undergo extracellular efflux via lysosomal exocytosis. Autophagy 17:690-705 (2021). PubMed: 32070194
    • Cahova M  et al. The increased activity of liver lysosomal lipase in nonalcoholic Fatty liver disease contributes to the development of hepatic insulin resistance. Biochem Res Int 2012:135723 (2012). PubMed: 21904679
    • Wang F  et al. Role of lysosomal acid lipase in the intracellular metabolism of LDL-transported dehydroepiandrosterone-fatty acyl esters. Am J Physiol Endocrinol Metab 295:E1455-61 (2008). WB ; Human . PubMed: 18796546

    レビューと Q&A

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    1-4 of 4 Abreviews or Q&A

    Western blot abreview for Anti-Lysosomal acid lipase antibody [9G7F12]

     Average
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Mouse Cell lysate - whole cell (J774 mouse macrophages, Human fibroblasts)
    Loading amount
    50000 cells
    Specification
    J774 mouse macrophages, Human fibroblasts
    Treatment
    lysis with LDS sample buffer, DTT, 70C for 10 min.
    Gel Running Conditions
    Reduced Denaturing
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · 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 Jun 15 2007

    Question

    I’m using HepG2 as positive control and I don’t get any clear band using the LIPA antibody. I do get a lot of unspecific bands across all my samples. I’ve attached a pic of one of my blots. In previous experiments I was getting a quite clear band for HepG2 around 45 kDa, but for the last four blots I got a lot of unspecific binding , even though I tried new HepG2 lysates. My B-tubulin control looks ok.

    In the blot I send you 6 is HepG2 and 5 is the negative control (mouse cells). As you see I’m getting pretty much the same bands.

    Is there an isolated lipa protein that I could use as a control?

    Thank you very much for your help.

    Best wishes,

    Read More
    Answer

    Thank you for contacting us.

    We do have LIPA protein available in catalogue. The ID is ab152501; https://www.abcam.com/lysosomal-acid-lipase-protein-tagged-ab152501.html

    I have checked the image you have kindly provided, there is indeed bands at 45kDa and ˜65 kDa in HepG2 lane. Please also be aware that there are two isoforms of this protein, one has predicted molecular weight 39kDa and other 45kDa and due to heavy glycosylation the observed band size will be higher than 39 and 45. In HepG2 cells line the two bands at ˜45kDa and ˜65 kDa seems correspond to two isoforms, (http://www.uniprot.org/uniprot/P38571) so further tests about the identity of these isoform are recommended.

    Please also cross check the negative LIPA protein expression in publications in mouse cells.

    Though you have used the same antibody in past however we should expect some lot to lot variation so we always recommend our customers trying different dilutions of primary or secondary antibodies. The other important step is lysis of the cells, changing lysis buffer(ice cold) sometime helps. Pleas elaos make sure the lysis buffer contains protease inhibitors.

    I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

    Read More

    Question

    Product code: 36597
    Lot number: GR46581-1
    Inquiry: Hello, I've just purchased a new antibody and I can't get my western blot to work. The previous tube of the antibody was working ok and I didn't change anything in the protocol. I was wondering if you have a LIPA positive control that I can use to verify that it is not a problem with my samples. Thank you very much.

    Read More
    Answer

    Thank you for your email. I am sorry to hear that you have been experiencing problems with this antibody.

    This antibody has been tested with HepG2 cells and lung tissue sections; these samples will form a good positive control. Please find attached links to Human Protein Atlas; the database shows different tissue sections and cell lines in which this protein is highly expressed.

    http://www.proteinatlas.org/ENSG00000107798

    I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

    Read More

    Question

    BATCH NUMBER -- NOT SPECIFIED -- ORDER NUMBER 169514 DESCRIPTION OF THE PROBLEM Multiple bands, wrong size SAMPLE recombinant protein PRIMARY ANTIBODY Lysosomal acid lipase antibody [9G7F12] from abcam DETECTION METHOD ecl POSITIVE AND NEGATIVE CONTROLS USED negative control ANTIBODY STORAGE CONDITIONS -20 alliquot SAMPLE PREPARATION western blotting sample buffer clap 5 min boiling AMOUNT OF PROTEIN LOADED 30 ug ELECTROPHORESIS/GEL CONDITIONS 10% sds-page TRANSFER AND BLOCKING CONDITIONS general agent SECONDARY ANTIBODY GAM HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 1 HAVE YOU RUN A "NO PRIMARY" CONTROL? Yes ADDITIONAL NOTES Please give me more info about the 'Truncated LAL recombinant protein'. What's the antigen you use to produce this antibody, is it LAL or a part of it? (If yes, which part?) And where was the protein in your demonstrating picture truncated from?

    Read More
    Answer

    Dear Wei, Thank you for your patience. Regarding ab36597 (Mouse monoclonal [9G7F12] to Lysosomal acid lipase), information on the immunogen is proprietary. I am sorry that I can not provide additional information to you at this time. Please let me know if my suggestions improve your WB results. I look forward to your reply.

    Read More

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