33-5500

antibody from Invitrogen Antibodies

Targeting: SPARC BM-40, ON, ONT

Provider product page for 33-5500

Western blot

ELISA

Immunohistochemistry

Flow cytometry

Other assay

Antibody data

Antibody Data
Antigen structure
References [28]
Comments [0]
Validations
Other assay [6]

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Product number: 33-5500 - Provider product page
Provider: Invitrogen Antibodies
Product name: SPARC Monoclonal Antibody (ON1-1)
Antibody type: Monoclonal
Antigen: Other
Description: Source: Monoclonal antibody was obtained by fusing the mouse myeloma cell-line P3-X63-Ag8-U1 with spleen cells of BALB/c mouse after immunization with bovine bone osteonectin (SPARC). The monoclonal antibody was harvested from ascitic fluid. Specificity: This antibody reacts with both bone derived and platelet derived Osteonectin (ELISA). This antibody does not react with non-activated platelet and reacts with thrombin-stimulated platelet (flow cytometric analysis). This antibody reacts with human and bovine Osteonectin (Western blot analysis). This antibody cross reacts with rabbit and porcine Osteonectin. (ELISA) Storage: 4°C. This product does not contain preservative. The stock solution (2.0 mg/mL) should be stored in aliquots at -20°C for 1 year, or should be stored at 4°C for 6 months after adding 0.1% sodium azide. Avoid repeated freeze-thaw cycles. Reconstitution: Dissolve the lyophilized antibody in 50 µL of distilled water (final concentration: 2.0 mg/mL). This solution can be used as a stock solution. If dilution is necessary for your application, dilute the stock solution with the following Dilution solution just prior to use. When the entire amount of antibody is to be used over a short time period, it may be dissolved directly in 500 µL or more of the Dilution solution. Be sure to store the antibody at a minimum concentration of 2.0 mg/mL. A lower antibody concentration may result in decreased stability. Reconstituted antibody solution should contain 0.1% sodium azide as a preservative when stored at 4°C.
Reactivity: Human, Bovine, Porcine, Rabbit
Host: Mouse
Isotype: IgG
Antibody clone number: ON1-1
Vial size: 100 µg
Concentration: 2.0 mg/mL
Storage: 4° C

SPARC protein structure - 33-5500 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 2 ( a ) Adenoma; ( b ) adenoma with epithelial misplacement--epithelial misplacement in the submucosa is marked by asterix *; ( c ) adenoma with early carcinoma; invasive carcinoma in the submucosa is marked by asterix *. HE staining, orig. magnification 4x. FN 1: Mild positive reaction in the stroma, mostly in capillaries, in adenoma ( d ). More intensive reaction in the stroma in epithelial misplacement ( e ). Focal reaction in carcinoma cells and in the stroma of invasive carcinoma ( f ). SPARC: Very mild positive reaction in epithelial cells in adenoma ( g ). Mild positive reaction in epithelial cells and focal positive reaction in the stroma in epithelial misplacement ( h ). Mild positive reaction in carcinoma cells and strong positive reaction in the stroma of invasive carcinoma ( i ). DCN: Mild positive reaction in epithelial cells in adenoma ( j ). Mild positive reaction in epithelial cells and strong positive reaction in the stroma in epithelial misplacement ( k ). No positive reaction in carcinoma cells and strong positive reaction in the stroma of invasive carcinoma ( l ). Immunohistochemistry, orig. magnification 10x.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Fig. 1 ECM related molecules are increased in CAF and selectively released in their conditioned medium: a Western Blot analysis of whole lysates of NF and CAF showing the protein levels of the three molecules (COL10A1, SPARC and COL11A1,) in CAF and NF cell cultures. The histograms show relative quantification by densitometric analysis normalized to Vinculin. b Western Blot analysis for the presence of the proteins COL10A1, SPARC and COL11A1 released in the CM of CAF or NF. The histograms show the densitometric analysis (in arbitrary Units) after normalization with respect to total number of cells. The difference was considered significant at p -value < 0.05

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 1 Expression of SPARC in biopsy samples of patients with PCa. ( a ) Representative images of IHC against SPARC in TMAs of samples of BPH and PCa of low, intermediate, and high GS. Lower row corresponds to a magnification of the upper images. Scale bars = 100 mum. ( b ) Number of biopsy samples analyzed per group. ( c ) Quantification of DAB signal. The box-plots depict the intensity SPARC signal in each group. Dots outside the box-plot depict the outliers. * P

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 2 Basal expression, silencing, and overexpression of SPARC in PCa cell lines. ( a ) Representative image of western blot against SPARC in PCa cell lines 22Rv1, LNCaP, DU145 and PC3. ( b ) RT-qPCR of SPARC in PCa cell lines. DeltaDeltaCt was obtained after normalizing to PUM1 and PC3 cell lines ( n = 3). (c and d) PC3 cells were stable transduced with shRNA against SPARC (ShSPARC) or scramble (ShScr). LNCaP cells were stable transduced with SPARC sequence (SPARC-HA) or an empty vector (). Parental cells (Input) were used as control. ( c ) Representative images of western blot against SPARC in the different cell lines produced. Quantification of optic density was normalized to beta-actin and parental cells, numbers show median ( n = 3). ( d ) SPARC mRNA expression assessed through RT-qPCR ( n = 3); *** P

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Fig 3 Serum Validation. A) Percent recovery for three representative biomarkers. Matrix effects can either dampen signal (CA1, PAP) or inflate signal (SPARC). Ideal dilution factors were dependent on sensitivity of the assay and reference range of the biomarker ( Table 1 ). Black dashed line indicates 100% spike recovery. B) Bland-Altman plot [ 38 ] validating specimen integrity of a subset (N = 35) of the clinical samples with high sample volumes. Red dashed line indicates 95% confidence interval.