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    sparc-antibody-epr20121-ab207743.pdf

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遺伝子ノックアウト細胞株 検証済リコンビナントRabMAb

Anti-SPARC 抗体 [EPR20121] (ab207743)

  • Datasheet
  • Certificate of Compliance
Reviews (2) Submit a question References (3)

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Western blot - Anti-SPARC antibody [EPR20121] (ab207743)
  • Flow Cytometry (Intracellular) - Anti-SPARC antibody [EPR20121] (ab207743)
  • Western blot - Anti-SPARC antibody [EPR20121] (ab207743)
  • Western blot - Anti-SPARC antibody [EPR20121] (ab207743)
  • Immunoprecipitation - Anti-SPARC antibody [EPR20121] (ab207743)
  • Anti-SPARC antibody [EPR20121] (ab207743)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR20121] to SPARC
  • Suitable for: WB, IP, Flow Cyt (Intra)
  • Knockout validated
  • Reacts with: Human

Conjugates logo Related conjugates and formulations

Carrier Free

こちらの製品もご検討ください

タンパク質
Recombinant human SPARC protein (ab107952)
二次抗体
Product image
Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
一次抗体
Product image
Anti-Contactin 1 antibody (ab66265)

関連製品

製品の概要

  • 製品名

    Anti-SPARC antibody [EPR20121]
    SPARC 一次抗体 製品一覧
  • 製品の詳細

    Rabbit monoclonal [EPR20121] to SPARC
  • 由来種

    Rabbit
  • アプリケーション

    適用あり: WB, IP, Flow Cyt (Intra)more details
  • 種交差性

    交差種: Human
  • 免疫原

    Recombinant full length protein. This information is proprietary to Abcam and/or its suppliers.

  • ポジティブ・コントロール

    • WB: SH-SY5Y and A-375 whole cell lysates; Human bone giant cell tumor lysate; Human serum and brain lysate. Flow Cyt (intra): A-375 cells. IP: A-375 whole cell lysate.
  • 特記事項

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

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 long term. Avoid freeze / thaw cycle.
  • バッファー

    pH: 7.2
    Preservative: 0.01% Sodium azide
    Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
  • Concentration information loading...
  • 精製度

    Protein A purified
  • ポリ/モノ

    モノクローナル
  • クローン名

    EPR20121
  • アイソタイプ

    IgG
  • 研究分野

    • Cell Biology
    • Cell Cycle
    • Cell Cycle Inhibitors
    • Other
    • Cardiovascular
    • Angiogenesis
    • Adhesion / ECM
    • Extracellular Matrix
    • Signal Transduction
    • Protein Phosphorylation
    • Tyrosine Kinases
    • Receptor Tyrosine Kinases
    • Signal Transduction
    • Cytoskeleton / ECM
    • Extracellular Matrix
    • Structures
    • Bone
    • Stem Cells
    • Lineage Markers
    • Endoderm
    • Stem Cells
    • Mesenchymal Stem Cells
    • Osteogenesis
    • Cancer
    • Signal transduction
    • Protein phosphorylation
    • Tyrosine kinases
    • Receptor tyrosine kinases
    • Developmental Biology
    • Lineage specification
    • Endoderm

関連製品

  • Alternative Versions

    • Anti-SPARC antibody [EPR20121] - Low endotoxin, Azide free (ab223156)
    • Anti-SPARC antibody [EPR20121] - BSA and Azide free (ab271980)
  • Compatible Secondaries

    • Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Recombinant Protein

    • Recombinant human SPARC protein (ab107952)
  • Related Products

    • VeriBlot for IP Detection Reagent (HRP) (ab131366)

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ab207743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
1/1000. Detects a band of approximately 43 kDa (predicted molecular weight: 35 kDa).
IP
1/40.
Flow Cyt (Intra)
1/70.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

特記事項
WB
1/1000. Detects a band of approximately 43 kDa (predicted molecular weight: 35 kDa).
IP
1/40.
Flow Cyt (Intra)
1/70.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

ターゲット情報

  • 機能

    Appears to regulate cell growth through interactions with the extracellular matrix and cytokines. Binds calcium and copper, several types of collagen, albumin, thrombospondin, PDGF and cell membranes. There are two calcium binding sites; an acidic domain that binds 5 to 8 Ca(2+) with a low affinity and an EF-hand loop that binds a Ca(2+) ion with a high affinity.
  • 配列類似性

    Belongs to the SPARC family.
    Contains 1 EF-hand domain.
    Contains 1 follistatin-like domain.
    Contains 1 Kazal-like domain.
  • 発生段階

    Expressed at high levels in tissues undergoing morphogenesis, remodeling and wound repair.
  • 細胞内局在

    Secreted > extracellular space > extracellular matrix > basement membrane. In or around the basement membrane.
  • Target information above from: UniProt accession P09486 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: 6678 Human
    • Omim: 182120 Human
    • SwissProt: P09486 Human
    • Unigene: 111779 Human
    • Unigene: 708558 Human
    • 別名

      • AA517111 antibody
      • Basement membrane protein 40 antibody
      • Basement-membrane protein 40 antibody
      • BM 40 antibody
      • BM-40 antibody
      • BM40 antibody
      • Cysteine rich protein antibody
      • hm:zeh0062 antibody
      • MGC128090 antibody
      • OI17 antibody
      • ON antibody
      • Osteonectin antibody
      • Secreted acidic cystein rich glycoprotein antibody
      • Secreted protein acidic and cysteine rich antibody
      • Secreted protein acidic and rich in cysteine antibody
      • Secreted protein acidic cysteine rich (osteonectin) antibody
      • Secreted protein acidic cysteine rich antibody
      • SPARC antibody
      • SPRC antibody
      • SPRC_HUMAN antibody
      see all

    画像

    • Western blot - Anti-SPARC antibody [EPR20121] (ab207743)
      Western blot - Anti-SPARC antibody [EPR20121] (ab207743)
      All lanes : Anti-SPARC antibody [EPR20121] (ab207743) at 1/1000 dilution

      Lane 1 : Wild-type HAP1 whole cell lysate
      Lane 2 : SPARC knockout HAP1 whole cell lysate

      Lysates/proteins at 20 µg per lane.

      Predicted band size: 35 kDa
      Observed band size: 35 kDa



      Lanes 1 - 2: Merged signal (red and green). Green - ab207743 observed at 35 kDa. Red - loading control, ab130007, observed at 130 kDa.

      ab207743 was shown to recognize SPARC in wild-type HAP1 cells as signal was lost at the expected MW in SPARC knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and SPARC knockout samples were subjected to SDS-PAGE. Ab207743 and ab130007 (Mouse anti-Vinculin loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

    • Flow Cytometry (Intracellular) - Anti-SPARC antibody [EPR20121] (ab207743)
      Flow Cytometry (Intracellular) - Anti-SPARC antibody [EPR20121] (ab207743)

      Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed A-375 (Human malignant melanoma cell line) cells labeling SPARCwith ab207743 at 1/70 dilution (red) compared with a rabbit monoclonal IgG isotype control (ab172730; black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (Alexa Fluor® 488) at 1/2000 dilution was used as the secondary antibody. 

       

       

    • Western blot - Anti-SPARC antibody [EPR20121] (ab207743)
      Western blot - Anti-SPARC antibody [EPR20121] (ab207743)
      All lanes : Anti-SPARC antibody [EPR20121] (ab207743) at 1/1000 dilution

      Lane 1 : SH-SY5Y (Human neuroblastoma cell line from bone marrow) whole cell lysate
      Lane 2 : A-375 (Human malignant melanoma cell line) whole cell lysate
      Lane 3 : Human bone giant cell tumor lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      Lanes 1-2 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
      Lane 3 : VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/4000 dilution

      Predicted band size: 35 kDa
      Observed band size: 43 kDa why is the actual band size different from the predicted?



      Blocking/Dilution buffer: 5% NFDM/TBST.

      Exposure time: Lane 1: 30 seconds; Lane 2/3: 3 minutes.

      The observed molecular weight is consistent with what has been described in the literature (PMID: 18442048).

    • Western blot - Anti-SPARC antibody [EPR20121] (ab207743)
      Western blot - Anti-SPARC antibody [EPR20121] (ab207743)
      All lanes : Anti-SPARC antibody [EPR20121] (ab207743) at 1/1000 dilution

      Lane 1 : Human serum
      Lane 2 : Human brain lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/4000 dilution

      Predicted band size: 35 kDa
      Observed band size: 43 kDa why is the actual band size different from the predicted?


      Exposure time: 5 seconds


      Blocking/Dilution buffer: 5% NFDM/TBST.

    • Immunoprecipitation - Anti-SPARC antibody [EPR20121] (ab207743)
      Immunoprecipitation - Anti-SPARC antibody [EPR20121] (ab207743)

      SPARC was immunoprecipitated from 0.35 mg of A-375 (Human malignant melanoma cell line) whole cell lysate with ab207743 at 1/40 dilution.

      Western blot was performed from the immunoprecipitate using ab207743 at 1/1000 dilution.

      VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

      Lane 1: A-375 whole cell lysate, 10µg (Input).

      Lane 2: ab207743 IP in A-375 whole cell lysate.

      Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab207743 in A-375 whole cell lysate.

      Blocking and dilution buffer and concentration: 5% NFDM/TBST.

      Exposure time: 3 minutes.

    • Anti-SPARC antibody [EPR20121] (ab207743)
      Anti-SPARC antibody [EPR20121] (ab207743)

    プロトコール

    • Western blot protocols
    • Flow cytometry protocols
    • Immunoprecipitation protocols

    Click here to view the general protocols

    データシートおよび資料

    • Datasheet download

      Download

    Certificate of Compliance

    To download a Certificate of Compliance, please enter your Lot number below:

    参考文献 (3)

    ab207743 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。

    ab207743 は 3 報の論文で使用されています。

    • Pan K  et al. SPARC promotes pancreatic cancer cell proliferation and migration through autocrine secretion into the extracellular milieu. Oncol Lett 21:485 (2021). PubMed: 33968201
    • Hao P  et al. MicroRNA-211 suppresses prostate cancer proliferation by targeting SPARC. Oncol Lett 15:4323-4329 (2018). PubMed: 29541199
    • Zhang S  et al. hsa-miR-29c-3p regulates biological function of colorectal cancer by targeting SPARC. Oncotarget 8:104508-104524 (2017). PubMed: 29262657

    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

    1-2 of 2 Abreviews or Q&A

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-SPARC antibody [EPR20121]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Rat Tissue sections (nasal turbinates)
    Antigen retrieval step
    None
    Permeabilization
    Yes - 0.1% Triton X-100
    Specification
    nasal turbinates
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 20°C
    Fixative
    Formaldehyde
    Read More

    Catherine Heyward

    Verified customer

    投稿 Sep 13 2021

    Immunocytochemistry/ Immunofluorescence abreview for Anti-SPARC antibody [EPR20121]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (periodontal ligament cells)
    Permeabilization
    Yes - 0.1% Triton X-100
    Specification
    periodontal ligament cells
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 20°C
    Fixative
    Paraformaldehyde
    Read More

    Catherine Heyward

    Verified customer

    投稿 Sep 09 2021

    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
    For licensing inquiries, please contact partnerships@abcam.com

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