SPARC Antibody [Unconjugated]

Western blot shows lysates of C2C12 mouse myoblast cell line and mouse placenta tissue. PVDF membrane was probed with 0.2 µg/mL of Goat Anti-Mouse SPARC Antigen Affinity-purified Polyclonal Antibody (Catalog # AF942) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for SPARC at approximately 35-37 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

SPARC/Osteonectin was detected in immersion fixed frozen sections of mouse embryo (E15) using Mouse SPARC/ Osteonectin Antigen Affinity-purified Polyclonal Antibody (Catalog # AF942) at 1.7 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS008) and counterstained with hematoxylin (blue). Specific staining was localized to developing cartilage. View our protocol for Chromogenic IHC Staining of Frozen Tissue Sections.

SPARC was detected in immersion fixed C2C12 mouse myoblast cell line using Goat Anti-Mouse SPARC Antigen Affinity-purified Polyclonal Antibody (Catalog # AF942) at 5 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counterstained with DAPI (blue). Specific staining was localized to endoplasmic reticuli. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Balb/C-3T3 mouse fibroblast cell line was stained with Goat Anti-Mouse SPARC Polyclonal Antibody (Catalog # AF942, filled histogram) or Goat IgG control antibody (AB-108-C, open histogram), followed by Phycoerythrin-conjugated anti-Goat IgG (F0107). To facilitate intracellular staining, cells were fixed with Flow Cytometry Fixation Buffer (FC004) and permeabilized with Flow Cytometry Permeabilization/Wash Buffer I (FC005). Staining was performed using our Staining Intracellular Molecules protocol.

• Reactivity: Mu
• Applications: WB, IHC, CyTOF-ready, Flow, ICC/IF
• Clonality: Polyclonal
• Host: Goat
• Conjugate: Unconjugated
• Concentration: LYOPH

SPARC Antibody [Unconjugated] Summary

• Immunogen: Mouse myeloma cell line NS0-derived recombinant mouse SPARC/Osteonectin
  Ala18-Ile302
  Accession # P07214
• Specificity: Detects mouse SPARC in direct ELISAs and Western blots.
• Source: N/A
• Isotype: IgG
• Clonality: Polyclonal
• Host: Goat
• Gene: SPARC
• Purity Statement: Antigen Affinity-purified

Applications/Dilutions

• Dilutions:
  • CyTOF-ready
  • Immunocytochemistry 5-15 ug/mL
  • Immunohistochemistry 1-15 ug/mL
  • Intracellular Staining by Flow Cytometry 0.25 ug/10^6 cells
  • Western Blot 0.2 ug/mL

Packaging, Storage & Formulations

• Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  12 months from date of receipt, -20 to -70 degreesC as supplied. 1 month, 2 to 8 degreesC under sterile conditions after reconstitution. 6 months, -20 to -70 degreesC under sterile conditions after reconstitution.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
• Preservative: No Preservative
• Concentration: LYOPH
• Reconstitution Instructions: Reconstitute at 0.2 mg/mL in sterile PBS.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for SPARC Antibody [Unconjugated]

• Basement-membrane protein 40
• BM-40
• ONcysteine-rich protein
• Osteonectin
• Secreted protein acidic and rich in cysteine
• secreted protein, acidic, cysteine-rich (osteonectin)
• SPARC

Background

SPARC, an acronym for “secreted protein, acidic and rich in cysteine”, is also known as osteonectin or BM-40 (1-5). It is the founding member of a family of secreted matricellular proteins with similar domain structure. The 302 amino acid (aa), 43 kDa protein contains a 17 aa signal sequence, an N-terminal acidic region that binds calcium, a follistatin domain containing Kazal-like sequences, and a C-terminal extracellular calcium (EC) binding domain with two EF-hand motifs (1-5). Crystal structure shows that residues implicated in cell binding, inhibition of cell spreading and disassembly of focal adhesions cluster on one face of SPARC, while a collagen binding epitope and an N-glycosylation site are opposite this face (6). SPARC is produced by fibroblasts, capillary endothelial cells, platelets, and macrophages, especially in areas of tissue morphogenesis and remodeling (3, 7). SPARC shows context-specific effects, but generally inhibits adhesion, spreading and proliferation, and promotes collagen matrix formation (3-5). For endothelial cells, SPARC disrupts focal adhesions and binds and sequesters PDGF and VEGF (3-5). SPARC is abundantly expressed in bone, where it promotes osteoblast differentiation and inhibits adipogenesis (5, 8). SPARC is potentially cleaved by metalloproteinases, producing an angiogenic peptide that includes the copper-binding sequence KGHK (7). Paradoxically, SPARC is highly expressed in many tumor types, yet expression mainly decreases the likelihood of metastasis and confers sensitivity to chemotherapy and radiation (4, 9, 10). Stabilin-1, which is expressed on alternately activated macrophages, is the first SPARC receptor to be identified. It binds the SPARC EC domain and mediates endocytosis for degradation (11). Mature mouse SPARC shows 97%, 92%, 92%, 92%, and 83% aa identity with rat, human, dog, cow, and chick SPARC, respectively.

1. Lankat-Buttgereit, B. et al. (1988) FEBS Lett. 236:352.
2. McVey, J.H. et al. (1988) J. Biol. Chem. 263:11111.
3. Sage, H. et al. (1989) J. Cell Biol. 109:341.
4. Framson, P.E. and E.H. Sage (2004) J. Cell. Biochem. 92:679.
5. Alford, A.I. and K.D. Hankenson (2006) Bone 38:749.
6. Hohenester, E. et al. (1997) EMBO J. 16:3778.
7. Sage, E.H. et al. (2003) J. Biol. Chem. 278:37849.
8. Delany, A.M. et al. (2003) Endocrinology 144:2588.
9. Koblinski, J.E. et al. (2005) Cancer Res. 65:7370.
10. Tai, I.T. et al. (2005) J. Clin. Invest. 115:1492.
11. Kzhyshkowska, J. et al. (2006) J. Immunol. 176:5825.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for SPARC Antibody (AF942)(40)

Publications using AF942

• Le, AD;Fu, M;Kumar, R;Zacharias, G;Garcia, ADR; Astrocyte modulation of synaptic plasticity mediated by activity-dependent Sonic hedgehog signaling bioRxiv : the preprint server for biology 2024-06-13 [PMID: 38915525] (Immunohistochemistry, Mouse)
• Niebergall, EB;Weekley, D;Mazur, A;Olszewski, NA;DeSchepper, KM;Radant, N;Vijay, AS;Risher, WC; Abnormal Morphology and Synaptogenic Signaling in Astrocytes Following Prenatal Opioid Exposure Cells 2024-05-14 [PMID: 38786059] (Western Blot, Rat)
• Laddach, A;Chng, SH;Lasrado, R;Progatzky, F;Shapiro, M;Erickson, A;Sampedro Castaneda, M;Artemov, AV;Bon-Frauches, AC;Amaniti, EM;Kleinjung, J;Boeing, S;Ultanir, S;Adameyko, I;Pachnis, V; A branching model of lineage differentiation underpinning the neurogenic potential of enteric glia Nature communications 2023-09-22 [PMID: 37737269] (IHC, Mouse)
• YS Song, N Jamali, CM Sorenson, N Sheibani Vitamin D Receptor Expression Limits the Angiogenic and Inflammatory Properties of Retinal Endothelial Cells Cells, 2023-01-16;12(2):. 2023-01-16 [PMID: 36672270] (Western Blot, Mouse)
• Luiza dos Santos Heringer, Julia Rios Carvalho, Julia Teixeira Oliveira, Bruna Texeira Silva, Domethila Mariano de Souza Aguiar dos Santos, Anna Lecticia Martinez Martinez Toledo et al. Altered excitatory and inhibitory neocortical circuitry leads to increased convulsive severity after pentylenetetrazol injection in an animal model of schizencephaly, but not of microgyria Epilepsia Open 9/1/2022 [PMID: 35808864]
• Christoph Ammer‐Herrmenau, Laurin Wolf, Syeda S. Nasrin, Iswarya Ramu, Roberta Roggiolani, Robert G. Goetze et al. Activity of acute pancreatitis is modified by secreted protein acidic and rich in cysteine ablation United European Gastroenterology Journal 7/1/2022 [PMID: 35699570]
• WW MacDonald, SS Swaminatha, JY Heo, A Castillejo, J Hsueh, BJ Liu, D Jo, A Du, H Lee, MH Kang, DJ Rhee Effect of SPARC Suppression in Mice, Perfused Human Anterior Segments, and Trabecular Meshwork Cells Investigative Ophthalmology & Visual Science, 2022-06-01;63(6):8. 2022-06-01 [PMID: 35671048] (IHC, Mouse)
• A Ghanemi, A Melouane, M Yoshioka, J St-Amand Secreted Protein Acidic and Rich in Cysteine (Sparc) KO Leads to an Accelerated Ageing Phenotype Which Is Improved by Exercise Whereas SPARC Overexpression Mimics Exercise Effects in Mice Metabolites, 2022-01-28;12(2):. 2022-01-28 [PMID: 35208200] (Western Blot, Mouse)
• Y Xie, AT Kuan, W Wang, ZT Herbert, O Mosto, O Olukoya, M Adam, S Vu, M Kim, D Tran, N Gómez, C Charpentie, I Sorour, TE Lacey, MY Tolstoruko, BL Sabatini, WA Lee, CC Harwell Astrocyte-neuron crosstalk through Hedgehog signaling mediates cortical synapse development Cell Reports, 2022-02-22;38(8):110416. 2022-02-22 [PMID: 35196485] (IHC, Mouse)
• RL Kosinsky, D Saul, C Ammer-Herr, WA Faubion, A Neesse, SA Johnsen USP22 Suppresses SPARC Expression in Acute Colitis and Inflammation-Associated Colorectal Cancer Cancers, 2021-04-10;13(8):. 2021-04-10 [PMID: 33920268] (IHC, Mouse)
• T Shoneye, AT Orrego, R Jarvis, Y Men, MSR Chiang, Y Yang Differential Proliferation and Maturation of Subcortical Astrocytes During Postnatal Development Front Neurosci, 2020-05-08;14(0):435. 2020-05-08 [PMID: 32457572] (IHC, Mouse)
• J Falero-Per, N Sheibani, CM Sorenson Bim expression modulates the pro-inflammatory phenotype of retinal astroglial cells PLoS ONE, 2020-05-04;15(5):e0232779. 2020-05-04 [PMID: 32365083] (Western Blot, Mouse)
• SD Praktiknjo, B Obermayer, Q Zhu, L Fang, H Liu, H Quinn, M Stoeckius, C Kocks, W Birchmeier, N Rajewsky Tracing tumorigenesis in a solid tumor model at single-cell resolution Nat Commun, 2020-02-20;11(1):991. 2020-02-20 [PMID: 32080185] (IHC, Mouse)
• Marie C. Hasselluhn, Lukas Klein, Melanie S. Patzak, Sören M. Buchholz, Philipp Ströbel, Volker Ellenrieder et al. Stromal Features of the Primary Tumor Are Not Prognostic in Genetically Engineered Mice of Pancreatic Cancer Cells 12/24/2019 [PMID: 31878349]
• I Ramu, SM Buchholz, MS Patzak, RG Goetze, SK Singh, FM Richards, DI Jodrell, B Sipos, P Ströbel, V Ellenriede, E Hessmann, A Neesse SPARC dependent collagen deposition and gemcitabine delivery in a genetically engineered mouse model of pancreas cancer EBioMedicine, 2019-10-06;0(0):. 2019-10-06 [PMID: 31597597] (IHC-P, Western Blot, Mouse)
• Methodios Ximerakis, Scott L. Lipnick, Brendan T. Innes, Sean K. Simmons, Xian Adiconis, Danielle Dionne et al. Single-cell transcriptomic profiling of the aging mouse brain Nature Neuroscience 10/1/2019 [PMID: 31551601]
• Ji Eun Park, Joonyoung Park, Yearin Jun, Yunseok Oh, Gongmi Ryoo, Yoo-Seong Jeong et al. Expanding therapeutic utility of carfilzomib for breast cancer therapy by novel albumin-coated nanocrystal formulation Journal of Controlled Release 5/1/2019 [PMID: 30954620]
• S Deckx, DM Johnson, M Rienks, P Carai, E Van Deel, J Van der Ve, KR Sipido, S Heymans, AP Papageorgi Extracellular SPARC increases cardiomyocyte contraction during health and disease PLoS ONE, 2019-04-01;14(4):e0209534. 2019-04-01 [PMID: 30933983] (ICC, Western Blot, Mouse)
• J Falero-Per, YS Song, Y Zhao, L Teixeira, CM Sorenson, N Sheibani Cyp1b1 expression impacts the angiogenic and inflammatory properties of liver sinusoidal endothelial cells PLoS ONE, 2018-10-29;13(10):e0206756. 2018-10-29 [PMID: 30372497] (Western Blot, Mouse)
• Lindsay T. McDonald, Michael R. Zile, Yuhua Zhang, An O. Van Laer, Catalin F. Baicu, Robert E. Stroud et al. Increased macrophage-derived SPARC precedes collagen deposition in myocardial fibrosis American Journal of Physiology-Heart and Circulatory Physiology 7/1/2018 [PMID: 29522370]
• EV Jones, Y Bernardine, JG Zarruk, S Chierzi, KK Murai SPARC and GluA1-Containing AMPA Receptors Promote Neuronal Health Following CNS Injury Front Cell Neurosci, 2018-02-01;12(0):22. 2018-02-01 [PMID: 29449802] (IHC-P, Western Blot, Mouse)
• M Farnoodian, CM Sorenson, N Sheibani PEDF Expression Affects the Oxidative and Inflammatory State of Choroidal Endothelial Cells Am. J. Physiol., Cell Physiol., 2018-01-10;0(0):. 1/10/2018 [PMID: 29351407]
• SF Gad, J Park, JE Park, GN Fetih, SS Tous, W Lee, Y Yeo Enhancing Docetaxel Delivery to Multidrug-Resistant Cancer Cells with Albumin-Coated Nanocrystals Mol. Pharm., 2018-01-29;0(0):. 1/29/2018 [PMID: 29341617]
• Jessica Wallingford, Angela L. Scott, Kelly Rodrigues, Laurie C. Doering Altered Developmental Expression of the Astrocyte-Secreted Factors Hevin and SPARC in the Fragile X Mouse Model Frontiers in Molecular Neuroscience 8/29/2017 [PMID: 28900386]
• L Morel, MSR Chiang, H Higashimor, T Shoneye, LK Iyer, J Yelick, A Tai, Y Yang Molecular and Functional Properties of Regional Astrocytes in the Adult Brain J. Neurosci., 2017-08-07;0(0):. 2017-08-07 [PMID: 28821665] (IHC, Mouse)
• Stabilin-1 is expressed in human breast cancer and supports tumor growth in mammary adenocarcinoma mouse model Oncotarget, 2016-05-24;7(21):31097-110. 2016-05-24 [PMID: 27105498] (IHC-Fr, Western Blot, Mouse)
• Tseng C, Kolonin M Proteolytic Isoforms of SPARC Induce Adipose Stromal Cell Mobilization in Obesity. Stem Cells, 2015-09-22;34(1):174-90. 2015-09-22 [PMID: 26381424] (IHC-P, Western Blot, Mouse)
• Mathsyaraja H, Thies K, Taffany D, Deighan C, Liu T, Yu L, Fernandez S, Shapiro C, Otero J, Timmers C, Lustberg M, Chalmers J, Leone G, Ostrowski M CSF1-ETS2-induced microRNA in myeloid cells promote metastatic tumor growth. Oncogene, 2014-09-22;34(28):3651-61. 2014-09-22 [PMID: 25241894] (Western Blot, Mouse)
• Sangaletti , Sabina, Tripodo , Claudio, Portararo , Paola, Dugo , Matteo, Vitali , Caterina, Botti , Laura, Guarnotta , Carla, Cappetti , Barbara, Gulino , Alessand, Torselli , Ilaria, Casalini , Patrizia, Chiodoni , Claudia, Colombo , Mario P Stromal niche communalities underscore the contribution of the matricellular protein SPARC to B-cell development and lymphoid malignancies. Oncoimmunology, 2014-06-05;3(0):e28989. 2014-06-05 [PMID: 25083326] (IHC, IHC-P, Mouse)
• Swarup S. Swaminathan, Dong-Jin Oh, Min Hyung Kang, Allan R. Shepard, Iok-Hou Pang, Douglas J. Rhee TGF-beta 2–Mediated Ocular Hypertension Is Attenuated in SPARC-Null Mice Investigative Opthalmology & Visual Science 7/1/2014 [PMID: 24906856]
• Stacey L. Thomas, Chad R. Schultz, Ezekiell Mouzon, William A. Golembieski, Reima El El Naili, Archanna Radakrishnan et al. Loss of Sparc in p 53‐null Astrocytes Promotes Macrophage Activation and Phagocytosis Resulting in Decreased Tumor Size and Tumor Cell Survival Brain Pathology 2015-07-01 [PMID: 24862407] (Immunohistochemistry, Western Blot, Transgenic Mouse)
• Stacey L. Thomas, Chad R. Schultz, Ezekiell Mouzon, William A. Golembieski, Reima El El Naili, Archanna Radakrishnan et al. Loss of Sparc in p 53‐null Astrocytes Promotes Macrophage Activation and Phagocytosis Resulting in Decreased Tumor Size and Tumor Cell Survival Brain Pathology 7/1/2015 [PMID: 24862407]
• Albrecht Neesse, Kristopher K Frese, Derek S Chan, Tashinga E Bapiro, William J Howat, Frances M Richards et al. SPARC independent drug delivery and antitumour effects of nab-paclitaxel in genetically engineered mice Gut 2014-06-01 [PMID: 24067278] (Immunohistochemistry, Western Blot, Mouse)
• Albrecht Neesse, Kristopher K Frese, Derek S Chan, Tashinga E Bapiro, William J Howat, Frances M Richards et al. SPARC independent drug delivery and antitumour effects of nab-paclitaxel in genetically engineered mice Gut 6/1/2014 [PMID: 24067278]
• Arnold SA, Rivera LB, Carbon JG, Toombs JE, Chang CL, Bradshaw AD, Brekken RA Losartan slows pancreatic tumor progression and extends survival of SPARC-null mice by abrogating aberrant TGF beta activation. PLoS ONE, 2012-02-14;7(2):e31384. 2012-02-14 [PMID: 22348081] (ICC, Mouse)
• Jones EV, Bernardinelli Y, Tse YC, Chierzi S, Wong TP, Murai KK Astrocytes control glutamate receptor levels at developing synapses through SPARC-beta-integrin interactions. J. Neurosci., 2011-03-16;31(11):4154-65. 2011-03-16 [PMID: 21411656] (IHC-Fr, Mouse)
• Schellings MW, Vanhoutte D, Swinnen M, Cleutjens JP, Debets J, van Leeuwen RE, d'Hooge J, Van de Werf F, Carmeliet P, Pinto YM, Sage EH, Heymans S Absence of SPARC results in increased cardiac rupture and dysfunction after acute myocardial infarction. J. Exp. Med., 2008-12-22;206(1):113-23. 2008-12-22 [PMID: 19103879] (Western Blot, Mouse, Rat)
• Faro-Trindade I, Cook PR A conserved organization of transcription during embryonic stem cell differentiation and in cells with high C value. Mol. Biol. Cell, 2006-04-19;17(7):2910-20. 2006-04-19 [PMID: 16624866] (ICC, Western Blot, Mouse)
• Nozaki M, Sakurai E, Raisler BJ, Baffi JZ, Witta J, Ogura Y, Brekken RA, Sage EH, Ambati BK, Ambati J Loss of SPARC-mediated VEGFR-1 suppression after injury reveals a novel antiangiogenic activity of VEGF-A. J. Clin. Invest., 2006-02-01;116(2):422-9. 2006-02-01 [PMID: 16453023] (Western Blot, Mouse)
• Sweetwyne MT, Brekken RA, Workman G, Bradshaw AD, Carbon J, Siadak AW, Murri C, Sage EH Functional analysis of the matricellular protein SPARC with novel monoclonal antibodies. J. Histochem. Cytochem., 2004-06-01;52(6):723-33. 2004-06-01 [PMID: 15150281] (IHC-P, Western Blot, Human)

Bioinformatics

• Gene Symbol: SPARC
• Uniprot:
  • P07214()