Mouse SPARC Antibody

R&D Systems | Catalog # AF942

R&D Systems

Key Product Details

Species Reactivity

Validated:

Mouse

Cited:

Human, Mouse, Rat, Transgenic Mouse

Applications

Validated:

Immunohistochemistry, Western Blot, Intracellular Staining by Flow Cytometry, Immunocytochemistry, CyTOF-ready

Cited:

Immunohistochemistry, Immunohistochemistry-Paraffin, Immunohistochemistry-Frozen, Western Blot, Flow Cytometry, Immunocytochemistry, Western Blo

Label

Unconjugated

Antibody Source

Polyclonal Goat IgG
View all SPARC Primary Antibodies »

Product Specifications

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.

Clonality

Polyclonal

Host

Goat

Isotype

IgG

Scientific Data Images for Mouse SPARC Antibody

Detection of Mouse SPARC antibody by Western Blot.

Detection of Mouse SPARC by Western Blot.

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 antibody in Mouse Embryo by Immunohistochemistry (IHC-Fr).

SPARC/Osteonectin in Mouse Embryo.

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 antibody in C2C12 Mouse Cell Line by Immunocytochemistry (ICC).

SPARC in C2C12 Mouse Cell Line.

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.

Detection of SPARC in Balb/C-3T3 Mouse Cell Line by Flow Cytometry.

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.
Detection of SPARC by Western Blot

Detection of SPARC by Western Blot

SPARC controls adiposity.(A) Schematic of experiments with inducible global Sparc KO mice. (B) Immunoblot analysis of SPARC protein in bone, VAT, and SAT in control Sparcfl/fl (Con), heterozygote (Sparcfl/+;CAG-CreER, Het iKO), and homozygote (Sparcfl/fl;CAG-CreER, Hom iKO) KO mice 6 weeks after tamoxifen injection. (C and D) Percentage of weight change of male (C) and female (D) littermate control (n = 10, 10), Het iKO (n = 10, 10), and Hom iKO (n = 5, 4) mice after tamoxifen injection. (E and F) Glucose tolerance test (GTT) (E), and insulin tolerance test (ITT) (F) of 14-month old (8 months after tamoxifen injection) male Con, Het iKO, and Hom iKO mice (n = 10, 10, 5). The blue star indicates statistical significance between Con and Het iKO mice, and the red star indicates statistical significance between Con and Hom iKO mice. Error bars represent the mean ± SEM. 2-way ANOVA test with Dunnett’s multiple comparisons test for adjusted P values (C–F) were performed for statistical analysis. *P < 0.05; **P < 0.01; ***P < 0.001. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/37781916), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of SPARC by Western Blot

Detection of SPARC by Western Blot

SPARC activates inflammation in macrophages via JNK signaling.(A) Human SPARC or mock vector was overexpressed in RAW 264.7 cells by transient transfection (0.5 and 2.5 μg) and Western-blot analyses of SPARC, JNK and p38 MAPK are shown. The experiment was repeated in triplicate and performed twice. (B) q-PCR analysis of Il1b, Tnf, Nos2, and Il6 in RAW 264.7 cells with SPARC or mock vector overexpression. (C) Primary BMDMs were treated with SPARC (5–60 minutes) and JNK, p65 NF-kappa B, and p38 MPAK were quantified by immunoblot analysis. (D) Representative immunoblot of p-p65 NF-kappa B in BMDMs pretreated with STAT1, p38, and JNK inhibitor and in presence of SPARC (20 μg/mL). (E) q-PCR analysis of Il1b, Tnf, Nos2, and Il6 in BMDMs pretreated with p38 or JNK inhibitor followed by SPARC treatment (20 μg/mL). (F) Primary BMDMs were transfected with JNK and STAT1 siRNA and immunoblot analysis was performed to quantify Pro-IL-1 beta protein levels. The experiment was repeated in triplicate and performed twice. Error bars represent the mean ± SEM. 2-tailed unpaired t tests (B) and 1-way ANOVA test with Bonferroni’s multiple comparisons test for adjusted P values (E) were performed for statistical analysis. *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/37781916), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of SPARC by Western Blot

Detection of SPARC by Western Blot

SPARC activates inflammation in macrophages via JNK signaling.(A) Human SPARC or mock vector was overexpressed in RAW 264.7 cells by transient transfection (0.5 and 2.5 μg) and Western-blot analyses of SPARC, JNK and p38 MAPK are shown. The experiment was repeated in triplicate and performed twice. (B) q-PCR analysis of Il1b, Tnf, Nos2, and Il6 in RAW 264.7 cells with SPARC or mock vector overexpression. (C) Primary BMDMs were treated with SPARC (5–60 minutes) and JNK, p65 NF-kappa B, and p38 MPAK were quantified by immunoblot analysis. (D) Representative immunoblot of p-p65 NF-kappa B in BMDMs pretreated with STAT1, p38, and JNK inhibitor and in presence of SPARC (20 μg/mL). (E) q-PCR analysis of Il1b, Tnf, Nos2, and Il6 in BMDMs pretreated with p38 or JNK inhibitor followed by SPARC treatment (20 μg/mL). (F) Primary BMDMs were transfected with JNK and STAT1 siRNA and immunoblot analysis was performed to quantify Pro-IL-1 beta protein levels. The experiment was repeated in triplicate and performed twice. Error bars represent the mean ± SEM. 2-tailed unpaired t tests (B) and 1-way ANOVA test with Bonferroni’s multiple comparisons test for adjusted P values (E) were performed for statistical analysis. *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/37781916), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of SPARC by Western Blot

Detection of SPARC by Western Blot

Adipocyte-derived SPARC controls macrophage inflammation.(A and B) Schematic and weight change of Con and Adip-KO mice with 21 weeks of HFD followed by 13 weeks of chow diet (n = 5, 5, respectively). (C) Body composition analysis of female Con and Adip-KO mice before and after diet change from HFD to chow diet (n = 4, 4, respectively). (D and E) q-PCR analysis of inflammatory gene (D) and components of inflammasome (E) in VAT macrophages (F4/80+) in obese mice switched to chow diet (n = 4, 4, respectively). (F) Inflammasome activation after pretreatment of SPARC protein for 24 hours following ATP (5mM) treatment with or without LPS (1 μg/mL) measured by caspase-1 Western blot analysis in cell lysate (lower) and supernatant (upper). Error bars represent the mean ± SEM. 2-tailed unpaired t tests were performed for statistical analysis. *P < 0.05; **P < 0.01; ***P < 0.001. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/37781916), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of SPARC by Western Blot

Detection of SPARC by Western Blot

Changes in synapse-related protein expression in POE astrocytes. (A) Western blot images reveal expression patterns of prominent synapse-associated proteins from the DIV10 vehicle and POE astrocyte lysates. Bottom row shows a representative band from one of the total protein blot images that was used for normalization purposes (full image shown in Figure S1). (B) Quantification of Western blots for proteins indicated in (A). Data shown as fold-change in band intensity values for POE astrocyte lysates compared to vehicle (n = 3–4 experimental replicates; unpaired t-test (unlabeled p values were not significant)). Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/38786059), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications for Mouse SPARC Antibody

Application
Recommended Usage

CyTOF-ready

Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.

Immunocytochemistry

5-15 µg/mL
Sample: Immersion fixed C2C12 mouse myoblast cell line

Immunohistochemistry

1-15 µg/mL
Sample: Immersion fixed frozen sections of mouse embryo (E15)

Intracellular Staining by Flow Cytometry

0.25 µg/106 cells
Sample: Balb/C-3T3 mouse embryonic fibroblast cell line fixed with paraformaldehyde and permeabilized with saponin

Western Blot

0.2 µg/mL
Sample: C2C12 mouse myoblast cell line and mouse placenta tissue

Reviewed Applications

Read 3 reviews rated 4.7 using AF942 in the following applications:

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Advanced Features

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Flow Cytometry

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Formulation

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.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: SPARC

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.

References

  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.

Long Name

Secreted Protein Acidic and Rich in Cysteine

Alternate Names

BM-40, Osteonectin

Entrez Gene IDs

6678 (Human); 20692 (Mouse)

Gene Symbol

SPARC

UniProt

P07214

Additional SPARC Products

Product Documents for Mouse SPARC Antibody

Certificate of Analysis

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Product Specific Notices for Mouse SPARC Antibody

For research use only

Citations for Mouse SPARC Antibody

Customer Reviews for Mouse SPARC Antibody (3)

4.7 out of 5
3 Customer Ratings
5 Stars
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4 Stars
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Customer Images

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  • Mouse SPARC Antibody
    Name: Emma Jones
    Application: Western Blot
    Sample Tested: Mouse hippocampal tissue
    Species: Mouse
    Verified Customer | Posted 02/02/2018
    Used at 1:3000 in 5% BSA and secondary goat-HRP antibody (R&D) at 1:5000
    Mouse SPARC Antibody AF942
  • Mouse SPARC Antibody
    Name: Anonymous
    Application: Western Blot
    Sample Tested: trabecular meshwork
    Species: Mouse
    Verified Customer | Posted 01/12/2018
    mouse SPARC was overexpressed by adeno-SPARC in mouse trabecular meshork (TM) cells.
    Mouse SPARC Antibody AF942
  • Mouse SPARC Antibody
    Name: Anonymous
    Application: Immunohistochemistry
    Sample Tested: Colon cancer tissue and Colon tissue
    Species: Mouse
    Verified Customer | Posted 05/18/2017
    Worked well in methacarn fixed tissues
    Mouse SPARC Antibody AF942

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