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Cell Biology Proteolysis / Ubiquitin Proteasome / Ubiquitin Proteasome
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Anti-Proteasome 20S LMP7 抗体 (ab3329)

  • Datasheet
Reviews (12)Q&A (3)References (40)

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Western blot - Anti-Proteasome 20S LMP7 antibody (ab3329)
  • Immunocytochemistry - Anti-Proteasome 20S LMP7 antibody (ab3329)
  • Western blot - Anti-Proteasome 20S LMP7 antibody (ab3329)
  • Immunocytochemistry - Anti-Proteasome 20S LMP7 antibody (ab3329)

Key features and details

  • Rabbit polyclonal to Proteasome 20S LMP7
  • Suitable for: WB, ICC
  • Reacts with: Human
  • Isotype: IgG

リコンビナント抗体で、ロット間での高い再現性を実現

Product image
Anti-Proteasome 20S LMP7 antibody [EPR14482(B)] (ab180606)
  • 異なるロット間での安定した再現性
  • 容易なスケールアップ
  • 評価試験による特異性の確認済み
  • 倫理基準に準拠 - アニマル・フリーの生産

製品の概要

  • 製品名

    Anti-Proteasome 20S LMP7 antibody
    Proteasome 20S LMP7 一次抗体 製品一覧
  • 製品の詳細

    Rabbit polyclonal to Proteasome 20S LMP7
  • 由来種

    Rabbit
  • アプリケーション

    適用あり: WB, ICCmore details
  • 種交差性

    交差種: Human
    交差が予測される動物種: Sheep, Cow
  • 免疫原

    Synthetic peptide corresponding to Human Proteasome 20S LMP7 aa 250-350.
    Database link: P28062

    Run BLAST with BLAST the sequence with ExPASy Run BLAST with BLAST the sequence with NCBI
  • 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

  • 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

    Constituents: 0.1% BSA, 99% PBS
  • Concentration information loading...
  • 精製度

    Immunogen affinity purified
  • ポリ/モノ

    ポリクローナル
  • アイソタイプ

    IgG
  • 研究分野

    • Cell Biology
    • Proteolysis / Ubiquitin
    • Proteasome / Ubiquitin
    • Proteasome
    • Neuroscience
    • Processes

関連製品

  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
  • Conjugation kits

    • PE / R-Phycoerythrin Conjugation Kit - Lightning-Link® (ab102918)
    • APC Conjugation Kit - Lightning-Link® (ab201807)
  • Isotype control

    • Rabbit IgG, polyclonal - Isotype Control (ChIP Grade) (ab171870)
  • Recombinant Protein

    • Recombinant Human Proteasome 20S LMP7 protein (denatured) (ab137150)

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ab3329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
WB (12)
Use a concentration of 1 - 3 µg/ml. Detects a band of approximately 20 kDa.
ICC
Use a concentration of 5 µg/ml.
特記事項
WB
Use a concentration of 1 - 3 µg/ml. Detects a band of approximately 20 kDa.
ICC
Use a concentration of 5 µg/ml.

ターゲット情報

  • 機能

    The proteasome is a multicatalytic proteinase complex which is characterized by its ability to cleave peptides with Arg, Phe, Tyr, Leu, and Glu adjacent to the leaving group at neutral or slightly basic pH. The proteasome has an ATP-dependent proteolytic activity. This subunit is involved in antigen processing to generate class I binding peptides. Replacement of PSMB5 by PSMB8 increases the capacity of the immunoproteasome to cleave model peptides after hydrophobic and basic residues. Acts as a major component of interferon gamma-induced sensitivity. Plays a key role in apoptosis via the degradation of the apoptotic inhibitor MCL1. May be involved in the inflammatory response pathway. In cancer cells, substitution of isoform 1 (E2) by isoform 2 (E1) results in immunoproteasome deficiency.
  • 関連疾患

    Defects in PSMB8 are the cause of JMP syndrome (JMPS) [MIM:613732]; also called joint contractures muscular atrophy microcytic anemia and panniculitis-induced lipodystrophy. JBTS1 is an autoinflammatory disorder characterized by childhood onset of joint stiffness and severe contractures of the hands and feet, erythematous skin lesions with subsequent development of severe lipodystrophy, and laboratory evidence of immune dysregulation. Accompanying features include muscle weakness and atrophy, hepatosplenomegaly, and microcytic anemia.
  • 配列類似性

    Belongs to the peptidase T1B family.
  • 発生段階

    Highly expressed in immature dendritic cells (at protein level).
  • 翻訳後修飾

    Autocleaved. The resulting N-terminal Thr residue of the mature subunit is responsible for the nucleophile proteolytic activity.
  • 細胞内局在

    Cytoplasm. Nucleus.
  • Target information above from: UniProt accession P28062 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • 参照データベース

    • Entrez Gene: 282013 Cow
    • Entrez Gene: 5696 Human
    • Entrez Gene: 100174905 Sheep
    • Omim: 177046 Human
    • SwissProt: Q3T112 Cow
    • SwissProt: P28062 Human
    • Unigene: 180062 Human
    • 別名

      • ALDD antibody
      • D6S216 antibody
      • D6S216E antibody
      • Large multifunctional peptidase 7 antibody
      • Large multifunctional protease 7 antibody
      • LMP 7 antibody
      • LMP7 antibody
      • Low molecular mass protein 7 antibody
      • Low molecular weight protein 7 antibody
      • Macropain subunit C13 antibody
      • MGC1491 antibody
      • Multicatalytic endopeptidase complex subunit C13 antibody
      • NKJO antibody
      • OTTHUMP00000062981 antibody
      • Protease component C13 antibody
      • Proteasome (prosome macropain) subunit beta type 8 antibody
      • Proteasome (prosome, macropain) subunit, beta type, 8 (large multifunctional peptidase 7) antibody
      • Proteasome beta 8 subunit antibody
      • Proteasome catalytic subunit 3i antibody
      • Proteasome component C13 antibody
      • Proteasome related gene 7 antibody
      • Proteasome subunit beta 5i antibody
      • Proteasome subunit beta 8 antibody
      • Proteasome subunit beta type 8 antibody
      • Proteasome subunit beta type antibody
      • Proteasome subunit beta type-8 antibody
      • Proteasome subunit beta-5i antibody
      • Proteasome subunit Y2 antibody
      • PSB8_HUMAN antibody
      • PSMB 8 antibody
      • PSMB5i antibody
      • PSMB8 antibody
      • Really interesting new gene 10 protein antibody
      • RING 10 antibody
      • RING10 antibody
      • Y2 antibody
      see all

    画像

    • Western blot - Anti-Proteasome 20S LMP7 antibody (ab3329)
      Western blot - Anti-Proteasome 20S LMP7 antibody (ab3329)
      All lanes : Anti-Proteasome 20S LMP7 antibody (ab3329) at 2 µg/ml

      Lane 1 : HeLa whole cell extracts
      Lane 2 : HeLa treated with IFN gamma (100ng/ml IFN gamma for 72h) whole cell extracts
      Lane 3 : U-937 whole cell extracts
      Lane 4 : Raji whole cell extracts
      Lane 5 : Ramos whole cell extracts

      Lysates/proteins at 30 µg per lane.

      Secondary
      All lanes : Goat anti-Rabbit IgG (H+L) HRP conjugate at 1/2500 dilution

      Observed band size: 20 kDa why is the actual band size different from the predicted?



      Detected by chemiluminescence.

    • Immunocytochemistry - Anti-Proteasome 20S LMP7 antibody (ab3329)
      Immunocytochemistry - Anti-Proteasome 20S LMP7 antibody (ab3329)

      Immunocytochemistry/ Immunofluorescence analysis of HeLa cells labeling Proteasome 20S LMP7 with ab3329 at 5µg/ml. The cells were fixed with 4% paraformaldehyde for 15 minutes, permeabilized with 0.1% Triton X-100 in TBS for 10 minutes, and blocked with 3% Blocker BSA in PBS for 15 minutes at room temperature. Cells were stained with or without Anti-Proteasome 20S LMP7 antibody (ab3329), at a concentration of 5µg/ml for 1 hour at room temperature, and then incubated with a Alexa Fluor® 488 goat anti-rabbit IgG secondary antibody at a dilution of 1/1000 for 1 hour s at room temperature (both panels, green). Nuclei (both panels, blue) were stained with Hoechst 33342 dye.

    • Western blot - Anti-Proteasome 20S LMP7 antibody (ab3329)
      Western blot - Anti-Proteasome 20S LMP7 antibody (ab3329)
      Western blot of proteasome 20S LMP7 from HeLa cell extract using ab3329.
    • Immunocytochemistry - Anti-Proteasome 20S LMP7 antibody (ab3329)
      Immunocytochemistry - Anti-Proteasome 20S LMP7 antibody (ab3329)
      ICC/IF image of ab3329 stained MCF7 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab3329, 5µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

    プロトコール

    • Flow cytometry protocols
    • Immunohistochemistry protocols
    • Immunocytochemistry & immunofluorescence protocols
    • Western blot protocols

    Click here to view the general protocols

    データシートおよび資料

    • Datasheet download

      Download

    参考文献 (40)

    ab3329 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。

    ab3329 は 40 報の論文で使用されています。

    • Menzel S  et al. ADP-Ribosylation Regulates the Signaling Function of IFN-?. Front Immunol 12:642545 (2021). PubMed: 33763084
    • Farini A  et al. Defective dystrophic thymus determines degenerative changes in skeletal muscle. Nat Commun 12:2099 (2021). PubMed: 33833239
    • Khilji MS  et al. The inducible ß5i proteasome subunit contributes to proinsulin degradation in GRP94-deficient ß-cells and is overexpressed in type 2 diabetes pancreatic islets. Am J Physiol Endocrinol Metab 318:E892-E900 (2020). PubMed: 32255680
    • Farini A  et al. PTX3 Predicts Myocardial Damage and Fibrosis in Duchenne Muscular Dystrophy. Front Physiol 11:403 (2020). PubMed: 32508664
    • Khilji MS  et al. The intermediate proteasome is constitutively expressed in pancreatic beta cells and upregulated by stimulatory, low concentrations of interleukin 1 ß. PLoS One 15:e0222432 (2020). PubMed: 32053590
    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-Proteasome 20S LMP7 antibody

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (THP-1 cells)
    Gel Running Conditions
    Reduced Denaturing (4 - 12%)
    Loading amount
    50000 cells
    Specification
    THP-1 cells
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 20°C
    Read More

    Abcam user community

    Verified customer

    投稿 May 21 2019

    Western blot abreview for Anti-Proteasome 20S LMP7 antibody

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (HTB-16 cells)
    Gel Running Conditions
    Reduced Denaturing (12%)
    Loading amount
    10 µg
    Treatment
    1ug INFgamma for 12hrs
    Specification
    HTB-16 cells
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C
    Read More

    Abcam user community

    Verified customer

    投稿 Oct 08 2018

    Western blot abreview for Anti-Proteasome 20S LMP7 antibody

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Rat Tissue lysate - whole (Skeletal muscle)
    Gel Running Conditions
    Reduced Denaturing (12%)
    Loading amount
    20 µg
    Specification
    Skeletal muscle
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C
    Read More

    Abcam user community

    Verified customer

    投稿 Feb 29 2016

    Western blot abreview for Anti-Proteasome 20S LMP7 antibody

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Mouse Tissue lysate - whole (Skeletal muscle)
    Gel Running Conditions
    Reduced Denaturing (12%)
    Loading amount
    20 µg
    Specification
    Skeletal muscle
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C
    Read More

    Abcam user community

    Verified customer

    投稿 Feb 29 2016

    Western blot abreview for Anti-Proteasome 20S LMP7 antibody

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Mouse Tissue lysate - whole (Skeletal muscle)
    Loading amount
    30 µg
    Specification
    Skeletal muscle
    Gel Running Conditions
    Reduced Denaturing (15% gel)
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C
    Read More

    DR. G.K Sakellariou

    Verified customer

    投稿 May 15 2013

    Western blot abreview for Anti-Proteasome 20S LMP7 antibody

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Recombinant protein (Recombinant LMP7-GST)
    Loading amount
    0.025 µg
    Specification
    Recombinant LMP7-GST
    Gel Running Conditions
    Reduced Denaturing (4-12% bis/tris)
    Blocking step
    Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
    Read More

    Abcam user community

    Verified customer

    投稿 May 31 2012

    Western blot abreview for Anti-Proteasome 20S LMP7 antibody

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Mouse Cell lysate - whole cell (dendritic cell)
    Loading amount
    1e+006 cells
    Specification
    dendritic cell
    Gel Running Conditions
    Non-reduced Denaturing
    Blocking step
    BSA as blocking agent for 16 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 4°C
    Read More

    Abcam user community

    Verified customer

    投稿 Jan 05 2010

    Question

    Dear Madam/Sir, Thank you very much for cooperation in resolving our problem with the antibodies! I am attaching a film image of the lysates from mouse endothelial cells (first 4 lanes) and Pericytes (lanes 5-8) blotted with Anti-20S core Ab from abcam. The higher band is about 52 kDa and the lower band is almost 5 kDa. I could not see any other band with size 20 kDa as expected. The smurf2 antibody is now working by increasing the concentration to 1/100 and washing 5X with TBST. I am attaching the ordering information as you requested. Please let us know when we could get the new batch of the anti- 20S Ab. Thank you, Best regards,

    Read More

    Abcam community

    Verified customer

    Asked on Jan 03 2012

    Answer

    Thank you for sending me this image. This data will be used to keep records of how our products are performing and for internal invetigation if needed. I noted that all tissue lysates that you have used have given the same non-specific signal. However I was wondering if you had tried a positive control lysate with your experiments. I ask this because according to SwissProt, in mouse tissue,this protreasome is expressed in spleen, thymus, lung, liver, heart and, at a very low level, in kidney.However it is not expressed in brain nor testis. My concern is that whilethe antibody may be recognizing non-specifically it may still be performing as there is not expression in the brain. This information is in the general annotations on this page: http://www.uniprot.org/uniprot/P28063 I would highly recommend using a control tissue lysate such as ab7937 Spleen (Mouse) Whole Cell Lysate - normal tissue ( https://www.abcam.com/Spleen-Mouse-Whole-Cell-Lysate-normal-tissue-ab7937.html). If you have any questions please do not hesitate to contact me.

    Read More

    Abcam Scientific Support

    Answered on Jan 03 2012

    Question

    Dear Madam/Sir, We have recently purchased an antibody (20s LMP7) #ab3329 Lot no. GR39959-4, from Abcam but it does not seem to be functional. Me and my colleague have tried it with both milk and BSA blocking solution and it yields two non-specific bands much higher and lower than expected (20kDa) size. We also have purchase the Smurf1 and Smurf2 antibodies. The Smurf1 antibody works good but Smurf2 (#ab38543) antibody weakly gives the expected band but it shows also lots of sand shaped spots on the membrane and washing for long time does not solve this problem. I wonder to know if we could get replacement for these two new purchases from another batch of antibody. Thank you for considering our case. Our shipping address is

    Read More

    Abcam community

    Verified customer

    Asked on Dec 28 2011

    Answer

    Thank you for taking time to send us your protocols. I am sorry to hear that this antibody is not providing satisfactory results. The details provided will enable us to investigate this case and will provide us with vital information for monitoring product quality. Having reviewed this case, I would like to offer some suggestions to help optimize the results from ab38543. Spotty/sandy looking blots are often the result of aggregates in the block or antibody. I would recommend when preparing the milk in TBST that you stir it with a stir bar for quite some time. This will help dissolve particulate matter that can stick to your blot. Filtering the milk will also remove any milk that has not dissolved. I would also suggest that you centrifuge the antibody before use and use the supernatant from the top should aggregates appear. You may use the supernatant at recommended dilutions without a reduction in quality. I would also recommend using a lower concentration for your secondary antibody. Some people have noted that rabbit secondaries used at high concentration may cause issues like this. To increase target signal, I would suggest using a higher concentration of ant- SMURF 2 antibody. This product has been used successfully at concentrations as high as 1:50. Should the suggestions not improve the results, please do let me know. In the event that a product is not functioning in the species and applications cited on the product datasheet (and the problem has been reported within 6 months of purchase), we would be pleased to provide a free of charge replacement, credit note, or refund. In regards to ab3329. I've seen that others have posted abreviews which have shown a higher non-specific band at about 66 kDa. Is this about the range of the higher band that you have reported? Do you have an image that you might send us? Would it also be possible to send me the original order number or purchase order for these products. If we need to replace them having that information will expedite the process. I hope this information is helpful, and I thank you for your cooperation.

    Read More

    Abcam Scientific Support

    Answered on Dec 28 2011

    Question

    Dear Madam/Sir, We have recently purchased an antibody (20s LMP7) #ab3329 from Abcam but it does not seem to be functional. Me and my colleague have tried it with both milk and BSA blocking solution and it yields two non-specific bands much higher and lower than expected (20kDa) size. We also have purchase the Smurf1 and Smurf2 antibodies. The Smurf1 antibody works good but Smurf2 (#ab38543) antibody weakly gives the expected band but it shows also lots of sand shaped spots on the membrane and washing for long time does not solve this problem. I wonder to know if we could get replacement for these two new purchases from another batch of antibody. Thank you for considering our case.

    Read More

    Abcam community

    Verified customer

    Asked on Dec 28 2011

    Answer

    Thank you for contacting us. I would like to ask you if you might send me the protocols used with these productsso that we can gather further information regarding the samples tested and the protocol used. Once we receive your response, we will look at the protocol and see if there are any suggestions we can make that may improve the results. This information will also allow us to investigate this case internally and initiate additional testing where necessary. If the product was purchased in the last six months and is being used according to our Abpromise, we would be happy to replace or refund the antibody. For our records could you please include the following details in your protocols for each antibody? What tissue type was used? How was the tissue prepared, including lysis buffer and conditions? Were the samples denatured? How much total protein was loaded per well? What were the running and transfer conditions and buffers? Did you use a loading control? Did you usethe recommended positive controls? What were the results of those controls? What blocking reagents did you use? You have mentioned that milk and BSA were used for ab3329, what were the concentrations of these in your block? What incubation times and temperatures did you use? What concentrations did you use the primary antibody concentration at? What incubation times, temperatures and reagents did you use? What was the secondary antibody? What were the concentrations, incubation times, temperatures and reagents used with this? Were different conditions tried, including different antibody concentrations and incubation times? What wash steps did you use? Which detection system did you use? I look forward to hearing form you soon.

    Read More

    Abcam Scientific Support

    Answered on Dec 28 2011

    1-10 of 15 Abreviews or Q&A

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    製品・研究分野別
    • 癌
    • 循環器
    • 細胞生物学
    • 発生
    • エピジェネティクス&核内情報
    • 免疫学
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    • 感染症
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    • ステムセル
    製品・タイプ別
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