Recombinant Anti-SPARC antibody [EPR20121] (ab207743)
![Western blot - Anti-SPARC antibody [EPR20121] (ab207743)](https://www.abcam.com/ps/products/207/ab207743/Images/ab207743-316719-anti-sparc-antibody-epr20121-western-blot.jpg "Western blot - 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
Related conjugates and formulations
Overview
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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, IP, Flow Cyt (Intra)more details
Unsuitable for: ICC/IF -
Species reactivity
Reacts with: Human
Does not react with: Mouse -
Immunogen
Recombinant full length protein. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- 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.
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General notes
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
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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Form
Liquid -
Storage instructions
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. -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading... -
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR20121 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Recombinant Protein
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Related Products
Applications
The Abpromise guarantee
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).
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| IP |
1/40.
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| Flow Cyt (Intra) |
1/70.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
| Notes |
|---|
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WB
1/1000. Detects a band of approximately 43 kDa (predicted molecular weight: 35 kDa). |
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IP
1/40. |
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Flow Cyt (Intra)
1/70. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
Target
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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
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Database links
- Entrez Gene: 6678 Human
- Omim: 182120 Human
- SwissProt: P09486 Human
- Unigene: 111779 Human
- Unigene: 708558 Human
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Alternative names
- AA517111 antibody
- Basement membrane protein 40 antibody
- Basement-membrane protein 40 antibody
see all
Images
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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 kDaLanes 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.
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![Flow Cytometry (Intracellular) - Anti-SPARC antibody [EPR20121] (ab207743)](https://www.abcam.com/ps/products/207/ab207743/Images/ab207743-270810-anti-sparc-antibody-epr20121-flow-cytometry.jpg)
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.
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![Western blot - Anti-SPARC antibody [EPR20121] (ab207743)](https://www.abcam.com/ps/products/207/ab207743/Images/ab207743-270812-anti-sparc-antibody-epr20121-western-blot.jpg)
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).
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![Western blot - Anti-SPARC antibody [EPR20121] (ab207743)](https://www.abcam.com/ps/products/207/ab207743/Images/ab207743-270811-anti-sparc-antibody-epr20121-western-blot.jpg)
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 secondsBlocking/Dilution buffer: 5% NFDM/TBST.
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![Immunoprecipitation - Anti-SPARC antibody [EPR20121] (ab207743)](https://www.abcam.com/ps/products/207/ab207743/Images/ab207743-270809-anti-sparc-antibody-epr20121-immunoprecipitation.jpg)
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.
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![Anti-SPARC antibody [EPR20121] (ab207743)](https://www.abcam.com/ps/products/207/ab207743/Images/ab207743-6-benefits-of-recombinant-antibodies.png)
Anti-SPARC antibody [EPR20121] (ab207743)
Protocols
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (3)
ab207743 has been referenced in 3 publications.
- 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