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リコンビナントRabMAb

Anti-SPARC 抗体 [EPR25122-122] - BSA and Azide free (ab290647)

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Immunocytochemistry/ Immunofluorescence - Anti-SPARC antibody [EPR25122-122] - BSA and Azide free (ab290647)
  • Immunocytochemistry/ Immunofluorescence - Anti-SPARC antibody [EPR25122-122] - BSA and Azide free (ab290647)
  • Western blot - Anti-SPARC antibody [EPR25122-122] - BSA and Azide free (ab290647)
  • Western blot - Anti-SPARC antibody [EPR25122-122] - BSA and Azide free (ab290647)
  • Western blot - Anti-SPARC antibody [EPR25122-122] - BSA and Azide free (ab290647)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SPARC antibody [EPR25122-122] - BSA and Azide free (ab290647)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SPARC antibody [EPR25122-122] - BSA and Azide free (ab290647)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SPARC antibody [EPR25122-122] - BSA and Azide free (ab290647)
  • Flow Cytometry (Intracellular) - Anti-SPARC antibody [EPR25122-122] - BSA and Azide free (ab290647)
  • Flow Cytometry (Intracellular) - Anti-SPARC antibody [EPR25122-122] - BSA and Azide free (ab290647)
  • Immunoprecipitation - Anti-SPARC antibody [EPR25122-122] - BSA and Azide free (ab290647)
  • Immunoprecipitation - Anti-SPARC antibody [EPR25122-122] - BSA and Azide free (ab290647)
  • Anti-SPARC antibody [EPR25122-122] - BSA and Azide free (ab290647)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR25122-122] to SPARC - BSA and Azide free
  • Suitable for: WB, Flow Cyt (Intra), IP, IHC-P, ICC/IF
  • Reacts with: Mouse, Rat

Conjugates logo Related conjugates and formulations

Unconjugated

製品の概要

  • 製品名

    Anti-SPARC antibody [EPR25122-122] - BSA and Azide free
    SPARC 一次抗体 製品一覧

  • 製品の詳細

    Rabbit monoclonal [EPR25122-122] to SPARC - BSA and Azide free

  • 由来種

    Rabbit

  • 特異性

    This antibody does not react with Rat species for IHC application. 

  • アプリケーション

    適用あり: WB, Flow Cyt (Intra), IP, IHC-P, ICC/IFmore details

  • 種交差性

    交差種: Mouse, Rat
    非交差種: Human

  • 免疫原

    Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.

  • ポジティブ･コントロール

    • WB: Mouse brain, heart, placenta tissue lysate. Rat brain, heart, placenta tissue lysates. C2C12, Neuro-2a, bEnd.3 and C6 whole cell lysate, untreated mouse and rat brain tissue lysate, mouse and rat brain tissue lysate treated with Protein Deglycosylation MIX II. Flow Cyt (intra): rat and mouse primary neuron cells. IP: Neuro-2a whole cell lysate, rat brain tissue. IHC-P: mouse hepatocellular, kidney, cerebrum tissues. ICC/IF: Permeabilized bEnd.3 and C6 cells.

  • 特記事項

    ab290647 is a carrier free version of ab290636. 

    Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.

    This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

    Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    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

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ab290647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
Use at an assay dependent concentration. Predicted molecular weight: 35 kDa.
Flow Cyt (Intra)
Use at an assay dependent concentration.
IP
Use at an assay dependent concentration.
IHC-P
Use at an assay dependent concentration.
ICC/IF
Use at an assay dependent concentration.
特記事項
WB
Use at an assay dependent concentration. Predicted molecular weight: 35 kDa.
Flow Cyt (Intra)
Use at an assay dependent concentration.
IP
Use at an assay dependent concentration.
IHC-P
Use at an assay dependent concentration.
ICC/IF
Use at an assay dependent concentration.

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.
  • Information by UniProt

  • 参照データベース

    • 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

    • Immunocytochemistry/ Immunofluorescence - Anti-SPARC antibody [EPR25122-122] - BSA and Azide free (ab290647)
      Immunocytochemistry/ Immunofluorescence - Anti-SPARC antibody [EPR25122-122] - BSA and Azide free (ab290647)

      Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized bEnd.3 (mouse brain endothelioma cell line) cells labelling SPARC with primary antibody anti-SPARC (ab290636) at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150081) secondary antibody at 1/1000 dilution (2.0 µg/mL). Confocal image showing cytoplasmic staining in bEnd.3 cells. Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) (ab195889) was used to counterstain tubulin at 1/200 dilution (2.5 µg/mL). The nuclear counter stain is DAPI (blue).
      The secondary antibody only control is : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 dilution (2.0 µg/mL).

      This data was developed using ab290636, the same antibody clone in a different buffer formulation.

    • Immunocytochemistry/ Immunofluorescence - Anti-SPARC antibody [EPR25122-122] - BSA and Azide free (ab290647)
      Immunocytochemistry/ Immunofluorescence - Anti-SPARC antibody [EPR25122-122] - BSA and Azide free (ab290647)

      Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized C6 (rat glial tumour cell line) cells labelling SPARC with primary antibody anti-SPARC (ab290636) at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150081) secondary antibody at 1/1000 dilution (2.0 µg/mL). Confocal image showing cytoplasmic staining in C6 cells. Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) (ab195889) was used to counterstain tubulin at 1/200 dilution (2.5 µg/mL). The nuclear counter stain is DAPI (blue).
      The secondary antibody only control is : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 dilution (2.0 µg/mL).

      This data was developed using ab290636, the same antibody clone in a different buffer formulation.

    • Western blot - Anti-SPARC antibody [EPR25122-122] - BSA and Azide free (ab290647)
      Western blot - Anti-SPARC antibody [EPR25122-122] - BSA and Azide free (ab290647)
      All lanes : Anti-SPARC antibody [EPR25122-122] (ab290636) at 1/1000 dilution

      Lane 1 : Mouse brain tissue lysate
      Lane 2 : Mouse heart tissue lysate
      Lane 3 : Mouse placenta tissue lysate
      Lane 4 : Rat brain tissue lysate
      Lane 5 : Rat heart tissue lysate
      Lane 6 : Rat placenta tissue lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

      Predicted band size: 35 kDa
      Observed band size: 40, 37 kDa why is the actual band size different from the predicted?



       This data was developed using ab290636, the same antibody clone in a different buffer formulation.

      Blocking and diluting buffer and concentration was 5% NFDM/TBST. 

      The expression profile/molecular weight observed is consistent with that described in the literature (PMID: 29449802; PMID: 9008236).
      The additional band is expected to be a SPARC cleavage product (PMID: 9008236).

      Exposure time: 

      Lane 2: 5.5 seconds
      Lane 1, 3-6: 3.25 seconds

    • Western blot - Anti-SPARC antibody [EPR25122-122] - BSA and Azide free (ab290647)
      Western blot - Anti-SPARC antibody [EPR25122-122] - BSA and Azide free (ab290647)
      All lanes : Anti-SPARC antibody [EPR25122-122] (ab290636) at 1/1000 dilution

      Lane 1 : C2C12 (mouse myoblast) whole cell lysate
      Lane 2 : bEnd.3 (mouse brain endothelioma) whole cell lysate
      Lane 3 : Neuro-2a (mouse neuroblastoma) whole cell lysate
      Lane 4 : C6 (rat glioma) whole cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

      Predicted band size: 35 kDa
      Observed band size: 40, 37 kDa why is the actual band size different from the predicted?



      This data was developed using ab290636, the same antibody clone in a different buffer formulation.

      Blocking and diluting buffer and concentration was 5% NFDM/TBST. 

      Exposure time: 3.25 seconds. 

      The expression profile/molecular weight observed is consistent with that described in the literature (PMID: 29449802; PMID: 9008236).
      The additional band is expected to be a SPARC cleavage product (PMID: 9008236).

    • Western blot - Anti-SPARC antibody [EPR25122-122] - BSA and Azide free (ab290647)
      Western blot - Anti-SPARC antibody [EPR25122-122] - BSA and Azide free (ab290647)
      All lanes : Anti-SPARC antibody [EPR25122-122] (ab290636) at 1/1000 dilution

      Lane 1 : Untreated mouse brain tissue lysate
      Lane 2 : Mouse brain tissue lysate treated with Protein Deglycosylation MIX II
      Lane 3 : Untreated rat brain tissue lysate
      Lane 4 : Rat brain tissue lysate treated with Protein Deglycosylation MIX II

      Lysates/proteins at 15 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

      Predicted band size: 35 kDa
      Observed band size: 40, 37 kDa why is the actual band size different from the predicted?


      Exposure time: 103 seconds


      This data was developed using ab290636, the same antibody clone in a different buffer formulation.

      Blocking and diluting buffer and concentration was 5% NFDM/TBST. 

      The expression profile/molecular weight observed is consistent with that described in the literature (PMID: 29449802; PMID: 9008236).
      SPARC is a glycosylated protein and can be deglycosylated by Protein Deglycosylation MIX II.
      The additional band is expected to be a SPARC cleavage product (PMID: 9008236).

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SPARC antibody [EPR25122-122] - BSA and Azide free (ab290647)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SPARC antibody [EPR25122-122] - BSA and Azide free (ab290647)

      This data was developed using ab290636, the same antibody clone in a different buffer formulation.

      Immunohistochemical analysis of paraffin-embedded mouse cerebrum tissue labelling SPARC with ab290636 at 1/15000 (0.041 µg/ml) followed by a ready to use LeicaDS9800 (BOND™ Polymer Refine Detection) . Positive staining on microglia of mouse cerebrum. The section was incubated with ab290636 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

      Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (BOND™ Polymer Refine Detection) .

      Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins. 

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SPARC antibody [EPR25122-122] - BSA and Azide free (ab290647)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SPARC antibody [EPR25122-122] - BSA and Azide free (ab290647)

      This data was developed using ab290636, the same antibody clone in a different buffer formulation.

      Immunohistochemical analysis of paraffin-embedded Mouse kidney tissue labelling SPARC with ab290636 at 1/15000 (0.041 µg/ml) followed by a ready to use LeicaDS9800 (BOND™ Polymer Refine Detection) . Positive staining on glomerulus and endothelium of mouse kidney. The section was incubated with ab290636 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.

      Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (BOND™ Polymer Refine Detection) .

      Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SPARC antibody [EPR25122-122] - BSA and Azide free (ab290647)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SPARC antibody [EPR25122-122] - BSA and Azide free (ab290647)

      This data was developed using ab290636, the same antibody clone in a different buffer formulation.

      Immunohistochemical analysis of paraffin-embedded mouse hepatocellular tissue labelling SPARC with ab290636 at 1/15000 (0.041 µg/ml) followed by a ready to use LeicaDS9800 (BOND™ Polymer Refine Detection) . Positive staining on interstitial cells of mouse hepatocellular carcinoma. The section was incubated with ab290636 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

      Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (BOND™ Polymer Refine Detection) .

      Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins. 

    • Flow Cytometry (Intracellular) - Anti-SPARC antibody [EPR25122-122] - BSA and Azide free (ab290647)
      Flow Cytometry (Intracellular) - Anti-SPARC antibody [EPR25122-122] - BSA and Azide free (ab290647)

      This data was developed using ab290636, the same antibody clone in a different buffer formulation.

      Intracellular flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Mouse primary neuron cell cells labelling SPARC with ab290636 at 1/600 dilution (0.1µg) (Right) compared with a Rabbit monoclonal IgG (ab172730) (Left) isotype control 0. A Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/2000 dilution was used as the secondary antibody.

    • Flow Cytometry (Intracellular) - Anti-SPARC antibody [EPR25122-122] - BSA and Azide free (ab290647)
      Flow Cytometry (Intracellular) - Anti-SPARC antibody [EPR25122-122] - BSA and Azide free (ab290647)

      This data was developed using ab290636, the same antibody clone in a different buffer formulation.

      Intracellular flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Rat primary neuron cells labelling SPARC with ab290636 at 1/600 dilution (0.1µg) (Right) compared with a Rabbit monoclonal IgG (ab172730) (Left). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/2000 dilution was used as the secondary antibody.

    • Immunoprecipitation - Anti-SPARC antibody [EPR25122-122] - BSA and Azide free (ab290647)
      Immunoprecipitation - Anti-SPARC antibody [EPR25122-122] - BSA and Azide free (ab290647)

      This data was developed using ab290636, the same antibody clone in a different buffer formulation.

      SPARC was immunoprecipitated from Neuro-2a (mouse neuroblastoma neuroblast) whole cell lysate with ab290636 at 1/30 dilution (2µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab290636 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

      Lane 1: Neuro-2a (mouse neuroblastoma neuroblast) whole cell lysate 10 µg

      Lane 2: ab290636 IP in Neuro-2a whole cell lysate 

      Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab290636 in Neuro-2a whole cell lysate

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

      Exposure time: 10 seconds

    • Immunoprecipitation - Anti-SPARC antibody [EPR25122-122] - BSA and Azide free (ab290647)
      Immunoprecipitation - Anti-SPARC antibody [EPR25122-122] - BSA and Azide free (ab290647)

      This data was developed using ab290636, the same antibody clone in a different buffer formulation.

      SPARC was immunoprecipitated from rat brain tissue lysate with ab290636 at 1/30 dilution (2µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab290636 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

      Lane 1: Rat brain tissue lysate 10 µg

      Lane 2: ab290636 IP in Rat brain tissue lysate

      Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab290636 in rat brain tissue lysate

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

      Exposure time: 10 seconds

    • Anti-SPARC antibody [EPR25122-122] - BSA and Azide free (ab290647)
      Anti-SPARC antibody [EPR25122-122] - BSA and Azide free (ab290647)

    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 (0)

    ab290647 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。

    ab290647 は論文での使用が確認できていません。

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