Overview

  • Product name

    Anti-SPARC antibody [EPR20121]
    See all SPARC primary antibodies
  • Description

    Rabbit monoclonal [EPR20121] to SPARC
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, Flow Cyt, IPmore details
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Recombinant full length protein within Human SPARC aa 1 to the C-terminus. The exact sequence is proprietary.
    Database link: P09486

  • Positive control

    • WB: SH-SY5Y and A-375 whole cell lysates; Human bone giant cell tumor lysate; Human serum and brain lysate. Flow Cyt: A-375 cells. IP: A-375 whole cell lysate.
  • General notes

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

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab207743 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
WB 1/1000. Detects a band of approximately 43 kDa (predicted molecular weight: 35 kDa).
Flow Cyt 1/70.

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

 

IP 1/40.

Target

  • Function

    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.
  • Sequence similarities

    Belongs to the SPARC family.
    Contains 1 EF-hand domain.
    Contains 1 follistatin-like domain.
    Contains 1 Kazal-like domain.
  • Developmental stage

    Expressed at high levels in tissues undergoing morphogenesis, remodeling and wound repair.
  • Cellular localization

    Secreted > extracellular space > extracellular matrix > basement membrane. In or around the basement membrane.
  • Information by UniProt
  • Database links

  • Alternative names

    • 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

Images

  • 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 cytometric analysis of 4% paraformaldehyde-fixed A-375 (Human malignant melanoma cell line) cells labeling SPARC with 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.

  • 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).

  • 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.

  • 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.

References

This product has been referenced in:

See all 2 Publications for this product

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