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QIA52 VEGF ELISA Kit, Mouse

VEGF ELISA Kit, Mouse : FDS (Fiches de données de sécurité), certificats d’analyse (CoA) et de qualité (CoQ), dossiers, brochures et autres documents disponibles.

Synonymes: Vascular Endothelial Growth Factor ELISA Kit, Vascular Permeability Factor ELISA Kit, VPF ELISA Kit

Detection Methods: Colorimetric
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      Aperçu

      Replacement Information

      Tableau de caractéristiques principal

      Detection Methods
      Colorimetric
      Description
      OverviewDetects and quantitates total, mouse VEGF.

      This product has been discontinued.
      Catalogue NumberQIA52
      Brand Family Calbiochem®
      SynonymsVascular Endothelial Growth Factor ELISA Kit, Vascular Permeability Factor ELISA Kit, VPF ELISA Kit
      Materials Required but Not Delivered• 2-20 µl, 20-200 µl and 200-1000 µl precision pipettors with disposable tips.
      • Wash bottle or multichannel dispenser for washing.
      • 500 ml graduated cylinder.
      • Deionized or distilled H2O.
      • Spectrophotometer capable of measuring absorbance in 96-well plates using dual wavelength of 450/540 nm or 450/570nm.
      • Polypropylene test tubes.
      References
      ReferencesClaffey, K.P., et al. 1996. Cancer Res. 56, 172.
      Joukov, V., et al. 1996. EMBO J. 16, 3898.
      Millauer, B., et al. 1996. Cancer Res. 56, 1615.
      Olofsson, B., et al. 1996. Proc. Natl. Acad. Sci. USA 93, 2576.
      Paavonen, K., et al. 1996. Circulation 93, 1079.
      Wei, M.H., et al. 1996. Hum. Genet. 97, 794.
      Asano, M., et al. 1995. Cancer Res. 55, 5296.
      Kolch, W., et al. 1995. Breast Cancer Res. Treat. 36, 139.
      Sato, T., et al. 1995. Nature 376, 70.
      Dumont, D.J., et al. 1994. Genes Dev. 8, 1897.
      Millauer, B., et al. 1994. Nature 367, 576.
      Takahashi, A., et al. 1994. Cancer Res. 54, 4233.
      Brown, L.F., et al. 1993. Cancer Res. 53, 4727.
      Brown, L.F., et al. 1993. Am. J. Pathol. 143, 1255.
      Millauer, B., et al. 1993. Cell 72, 835.
      Senger, D.R., et al. 1993. Cancer Metastasis Rev. 12, 303.
      Berse, B., et al. 1992. Mol. Biol. Cell 3, 211.
      Claffey, K.P., et al. 1992. J. Biol. Chem. 267, 16317.
      De Vries, C., et al. 1992. Science 255, 989.
      Plate, K.H., et al. 1992. Nature 359, 845.
      Terman, B., et al. 1992. Biochem. Biophys. Res. Commun. 187, 1579.
      Maglione, D., et al. 1991. Proc. Natl. Acad. Sci. USA 88, 9267.
      Matthews, W., et al. 1991. Proc. Natl. Acad. Sci. USA 88, 9026.
      Tischer, E., et al. 1991. J. Biol. Chem. 266, 11947.
      Shibuya, M., et al. 1990. Oncogene 5, 519.
      Keck, P.J., et al. 1989. Science 246, 1309.
      Leung, D.W., et al. 1989. Science 246, 1306.
      Dvorak, H.F., et al. 1979. J. Immunol. 122, 166.
      Product Information
      Detection methodColorimetric
      Form192 Tests
      Format96-well plate
      Kit containsTwo 96-well Coated Plates, Standard, VEGF Control, HRP Conjugate, Diluents, Duffers, Color Reagents, Stop Solution, and a user protocol.
      Applications
      Biological Information
      Assay range7.8-500 pg/ml
      Assay time4.5 h
      Sample TypeCell culture media, serum, tissue homogenates or mouse plasma
      Physicochemical Information
      Sensitivity3.0 pg/ml
      Dimensions
      Materials Information
      Toxicological Information
      Safety Information according to GHS
      Safety Information
      R PhraseR: 23/24-28-32-35-36/38-40-42/43-61

      Toxic by inhalation and in contact with skin.
      Very toxic if swallowed.
      Contact with acids liberates very toxic gas.
      Causes severe burns.
      Irritating to eyes and skin.
      Limited evidence of a carcinogenic effect.
      May cause sensitization by inhalation and skin contact.
      May cause harm to the unborn child.
      S PhraseS: 3/9/49-26-36/37/39-45

      Keep only in the original container in a cool, well ventilated place.
      In case of contact with eyes, rinse immediately with plenty of water and seek medical advice.
      Wear suitable protective clothing, gloves and eye/face protection.
      In case of accident or if you feel unwell, seek medical advice immediately (show the label where possible).
      Product Usage Statements
      Intended useThe Calbiochem® mouse VEGF ELISA Kit is a non-isotopic immunoassay for the in vitro quantitation of mouse vascular endothelial growth factor in cell culture media and mouse serum. This assay is for research use only and not for use in diagnostic or therapeutic procedures.
      Storage and Shipping Information
      Ship Code Blue Ice Only
      Toxicity Multiple Toxicity Values, refer to MSDS
      Storage +2°C to +8°C
      Storage ConditionsUpon arrival store the entire contents of the kit at 4°C.
      Do not freeze Ok to freeze
      Packaging Information
      Transport Information
      Supplemental Information
      Kit containsTwo 96-well Coated Plates, Standard, VEGF Control, HRP Conjugate, Diluents, Duffers, Color Reagents, Stop Solution, and a user protocol.
      Specifications
      Global Trade Item Number
      Référence GTIN
      QIA52 0

      Documentation

      VEGF ELISA Kit, Mouse Certificats d'analyse

      TitreNuméro de lot
      QIA52

      Références bibliographiques

      Aperçu de la référence bibliographique
      Claffey, K.P., et al. 1996. Cancer Res. 56, 172.
      Joukov, V., et al. 1996. EMBO J. 16, 3898.
      Millauer, B., et al. 1996. Cancer Res. 56, 1615.
      Olofsson, B., et al. 1996. Proc. Natl. Acad. Sci. USA 93, 2576.
      Paavonen, K., et al. 1996. Circulation 93, 1079.
      Wei, M.H., et al. 1996. Hum. Genet. 97, 794.
      Asano, M., et al. 1995. Cancer Res. 55, 5296.
      Kolch, W., et al. 1995. Breast Cancer Res. Treat. 36, 139.
      Sato, T., et al. 1995. Nature 376, 70.
      Dumont, D.J., et al. 1994. Genes Dev. 8, 1897.
      Millauer, B., et al. 1994. Nature 367, 576.
      Takahashi, A., et al. 1994. Cancer Res. 54, 4233.
      Brown, L.F., et al. 1993. Cancer Res. 53, 4727.
      Brown, L.F., et al. 1993. Am. J. Pathol. 143, 1255.
      Millauer, B., et al. 1993. Cell 72, 835.
      Senger, D.R., et al. 1993. Cancer Metastasis Rev. 12, 303.
      Berse, B., et al. 1992. Mol. Biol. Cell 3, 211.
      Claffey, K.P., et al. 1992. J. Biol. Chem. 267, 16317.
      De Vries, C., et al. 1992. Science 255, 989.
      Plate, K.H., et al. 1992. Nature 359, 845.
      Terman, B., et al. 1992. Biochem. Biophys. Res. Commun. 187, 1579.
      Maglione, D., et al. 1991. Proc. Natl. Acad. Sci. USA 88, 9267.
      Matthews, W., et al. 1991. Proc. Natl. Acad. Sci. USA 88, 9026.
      Tischer, E., et al. 1991. J. Biol. Chem. 266, 11947.
      Shibuya, M., et al. 1990. Oncogene 5, 519.
      Keck, P.J., et al. 1989. Science 246, 1309.
      Leung, D.W., et al. 1989. Science 246, 1306.
      Dvorak, H.F., et al. 1979. J. Immunol. 122, 166.

      Brochure

      Titre
      Angiogenesis and Tumor Metastasis Brochure and Technical Guide
      Kit SourceBook - 2nd Edition EURO
      Kits SourceBook - 2nd Edition GBP

      Citations

      Titre
    • Vivian Y. Shin, et al. (2005) Nicotine induces cyclooxygenase-2 and vascular endothelial growth factor receptor-2 in association with tumor-associated invasion and angiogenesis in gastric cancer. Molecular Cancer Research 3, 607-615.
      		•
      Protocole Utilisateur

      Revision28-September-2009 RFH
      SynonymsVascular Endothelial Growth Factor ELISA Kit, Vascular Permeability Factor ELISA Kit, VPF ELISA Kit
      Form192 Tests
      Format96-well plate
      Detection methodColorimetric
      Speciesmouse
      StorageUpon arrival store the entire contents of the kit at 4°C.
      Intended useThe Calbiochem® mouse VEGF ELISA Kit is a non-isotopic immunoassay for the in vitro quantitation of mouse vascular endothelial growth factor in cell culture media and mouse serum. This assay is for research use only and not for use in diagnostic or therapeutic procedures.
      BackgroundAngiogenesis, a term given to the growth of capillaries from pre-existing blood vessels, is found during embryonic development and is almost absent in the adult. While there are certain normal and disease states in the adult in which angiogenesis plays a role, most recent research work has concentrated on the role angiogenesis plays in tumor development and metastasis. Many factors have been shown to stimulate angiogenic growth including EGF, αFGF, βFGF, TGFα and VEGF to name but a few. VEGF (vascular endothelial growth factor) was firstly called VPF (vascular permeability factor) but now both names are used synonomously. VPF was originally described upon its ability to increase the permeability of the microvasculature to circulating macromolecules, such as plasma proteins. Later, VPF was shown to be a selective mitogenic factor for endothelial cells and was therefore given its second name, VEGF. The important role VEGF plays in tumorigenesis has been well documented. VEGF's effect in tumor development manifests itself in at least two ways: firstly as an endothelial growth factor stimulating endothelial cell proliferation and the formation of new blood vessels, and secondly as a vascular permeability factor generating fibrin networks which serve as a substratum for newly formed tissue. VEGF expression can be induced in many cell types by a number of stimuli including hypoxia, differentiation, growth factors and certain tumor promoters. VEGF's presence has been demonstrated in a wide range of tumors; including those from the central nervous system, gastrointestinal tract, kidney, and bladder. Another study has shown that VEGF can promote melanoma growth by inducing angiogenic growth and that constitutive VEGF expression dramatically promotes tumor colonization in the lung. This body of evidence, along with the multitude of other reports linking VEGF with tumor growth and metastasis, solidifies the notion that VEGF plays an important role in tumorigenesis. The gene for human VEGF has been localized, by fluorescence in situ hybridization, to chromosome 6p12. Human VEGF protein is a Mr 35,000-43,0000 dimeric polypeptide that can be expressed in different isoforms of 121, 165, 189 and 205 amino acids. The murine form of VEGF is closely related to its human counterpart, and murine VEGF mRNA is found in relatively low levels in normal murine tissues but is dramatically increased in models of differentiation and transformation. In addition to VEGF, three other members of the VEGF family have been identified, VEGF-B, VEGF-C and PIGF which all share sequence and structural homology to the primary family member, VEGF. The genes for human VEGF-B and -C have also been identified and are localized to 11q13 and 4q34, respectively. The additional three members of the VEGF family, just like VEGF itself, have been shown to play roles in angiogenesis. Two VEGF receptors have been identified, both being endothelial receptor tyrosine kinases (RTKs), and are called FLK1/KDR and FLT1. Glioblastoma growth in nude mice has been inhibited in vivo through the introduction of dominant-negative FLK1 mutants. In addition, inhibition of FLK1 signaling has been shown to inhibit the growth of many solid tumors. Finally, the injection of anti-VEGF antibody has demonstrated growth suppression activity in subcutaneously implanted fibrosarcoma cells. Taken together, these data suggest an important role for FLK1 in tumor angiogenesis. Three other RTKs have been identified, FLT4, TIE and TEK, of which TIE and TEK are essential for the development of proper vasculature. Mouse VEGF bioassays, based on the mitogenic activity of mouse VEGF are known to be tedious and non-specific for mouse VEGF. The Calbiochem® Mouse VEGF ELISA Kit (Cat. No. QIA52), measures mouse VEGF in tissue culture medium and mouse serum. The kit includes an Sf21-expressed mouse VEGF and anti-recombinant VEGF antibodies. The kit allows the quantitation of both natural and recombinant mouse VEGF, with the dose response curve for the natural version being parallel to that of the kit's recombinant standard curve. This demonstrates the kit's ability to quantitate natural mouse VEGF.
      Principles of the assayThe Calbiochem® mouse VEGF ELISA Kit is a "sandwich" enzyme immunoassay employing purified polyclonal antibodies. An affinity purified polyclonal antibody, specific for mouse VEGF protein, has been immobilized onto the surface of the plastic wells provided in the kit. The sample to be assayed (test samples, standards and controls) are pipetted into the wells and any mouse VEGF present binds to the capture antibody. Unbound material is washed away and a polyclonal, horseradish peroxidase (HRP)-conjugated anti-VEGF antibody is added to the wells. Following this incubation and a wash step, a chromogenic substrate is added to the wells. The horseradish peroxidase catalyses the conversion of the chromogenic substrate tetra-methylbenzidine (TMB) from a colorless solution to a blue solution (or yellow after the addition of stopping reagent), the intensity of which is proportional to the amount of mouse VEGF protein in the test sample. The colored reaction product is quantified using a spectrophotometer.

      Quantitation is achieved by the construction of a standard curve using known concentrations of mouse VEGF protein (provided lyophilized). By comparing the absorbance obtained from a sample containing an unknown amount of mouse VEGF protein with that obtained from the standards, the concentration of mouse VEGF protein in the test sample can be determined.
      Materials providedStandards should be assayed in duplicate. A standard curve must be performed on the same plate and at the same time as the samples. The mouse VEGF ELISA Kit is a two plate kit and therefore sufficient reagents are supplied to run 192 tests (including controls and standard curves).

      • Anti-VEGF (Mouse) Coated Plate (Kit Component No. JA1790): Two 96 well plates coated with anti-mouse VEGF polyclonal antibody. The plates are supplied as 12 removable strips containing 8 wells each.
      • MOUSE VEGF STANDARD (Kit Component No. JA1791): 2.5 ng of lyophilized recombinant mouse VEGF.
      • MOUSE VEGF CONJUGATE (Kit Component No. JA1792): 23 ml of a polyclonal antibody against mouse VEGF conjugated to horseradish peroxidase with preservatives.
      • MOUSE VEGF CONTROL (Kit Component No. JA1794): 1 vial of lyophilized mouse VEGF. The mouse VEGF control value obtained in the assay should be within the control value range stated on the vial label.
      • ASSAY DILUENT RD1N (Kit Component No. JA1795): 12.5 ml of buffer used to dilute standards and samples.
      • CALIBRATOR DILUENT (Kit Component No. JA1796): 2 vials (21 ml/vial) of buffer used to dilute mouse VEGF standard and mouse serum samples.
      • WASH BUFFER CONCENTRATE (Kit Component No. JA1797): 50 ml of a 25X concentrated solution of buffered surfactant.
      • COLOR REAGENT A (Kit Component No. JA1798): 12.5 ml of hydrogen peroxide.
      • COLOR REAGENT B (Kit Component No. JA1799): 12.5 ml of tetramethylbenzidine (TMB) chromogen.
      • STOP SOLUTION (Kit Component No. JA1813): 23 ml of a diluted hydrochloric acid.
      • PLATE SEALERS (Kit Component No. JA9373): To cover plates during incubations.
      Materials Required but not provided• 2-20 µl, 20-200 µl and 200-1000 µl precision pipettors with disposable tips.
      • Wash bottle or multichannel dispenser for washing.
      • 500 ml graduated cylinder.
      • Deionized or distilled H2O.
      • Spectrophotometer capable of measuring absorbance in 96-well plates using dual wavelength of 450/540 nm or 450/570nm.
      • Polypropylene test tubes.
      Precautions and recommendations• Store unopened kit at 4°C. Do not expose reagents to excessive light. Do not freeze any component.
      • Bring all reagents to room temperature before use.
      • The Stop Solution in the kit contains an acid solution. Exercise caution when handling this solution.
      • Use only the wells provided with the kit.
      • Do not mix reagents from different kits.
      • Do not mouth pipette or ingest any of the reagents.
      • The kit buffers and reagents contain preservatives. Do not make direct contact with these products.
      • Do not smoke, eat, or drink when performing the assay or in areas where samples/reagents are handled.
      • Samples may be contaminated with infectious agents. Do not ingest, expose to open wounds, or breathe aerosols. Dispose of samples properly.
      • Avoid adding reagents to the side of the wells. Add samples directly to the center of the well.
      • Add Stop Solution in same well order as the Substrate Solution.
      • Change pipette tips between different sample additions to avoid cross-contamination.
      PreparationSample Collection and Storage Cell Culture Medium: Centrifuge all samples to remove particulate material before assaying. Store all samples at ≤-20°C and avoid freeze/thaw cycles. Mouse serum: Note - grossly hemolyzed or lipemic samples are not suitable for analysis in this assay. Blood samples must be clotted for 2 h at room temperature or overnight at 4°C. Once clotted, samples are centrifuged at 2000 X g for 20 min. Carefully remove serum and assay. Store all serum samples at ≤-20°C and avoid freeze/thaw cycles. Sample Preparation For mouse serum samples it is recommended that the samples be diluted 1:5 in Calibrator Diluent RD5T (e.g. 50 µl sample mixed with 200 µl Calibrator Diluent).
      Reagent preparationNote: Bring all reagents to room temperature before use. Mouse VEGF Control: Reconstitute the control by adding 1ml of deionized or distilled water. The control is assayed undiluted. For repeat use of the control, it should be aliquoted and stored at ≤-20°C for up to 1 month (provided this is still within the shelf life stated on the kit). Avoid freeze/thaw cycles of the control. Wash Buffer: Warm the Wash Buffer Concentrate to room temperature before use (this will dissolve any crystals which may be present in the concentrate). Mix 25 ml of Wash Buffer Concentrate with 600 ml of deionized or distilled water to provide enough 1X wash buffer (625 ml) for one plate. The diluted Wash Buffer may be stored at 4°C for up to 1 month. Substrate Solution: Active Substrate Equal Volumes of Solution is produced by adding Color Reagents A and B together. 100 µl of substrate solution is required per well. Color Reagents A and B should be mixed together within 15 min of use. Mouse VEGF Standard: Reconstitute the Mouse VEGF Standard with 5 ml of Calibrator Diluent RD5T and allow to mix gently for 5 min before making serial dilutions. This reconstitution makes a stock solution of 500 pg/ml. For repeat use of the standard, it should be aliquoted and stored at ≤-20°C for up to 1 month. Avoid freeze/thaw cycles of the standard. To make serial dilutions of the standard, obtain six tubes and label them 250, 125, 62.5, 31.2, 15.6 and 7.8 pg/ml. Add 200 µl of Calibrator Diluent into each tube. Undiluted reconstituted standard serves as the top point in the standard curve (500 pg/ml) and Calibrator Diluent alone acts as the low point (0 pg/ml). Remove 200 µl from the undiluted, reconstituted standard vial (500 pg/ml) and add it to the first tube (250 pg/ml). Mix gently. Remove 200 µl from the first tube (250 pg/ml) and add it to the second tube (125 pg/ml) and mix gently. Repeat this procedure until you reach the sixth tube (7.8 pg/ml). Any remaining serially diluted standard should be discarded after one use. Note: do not discard the undiluted, reconstituted standard vial (500 pg/ml) - this should be aliquoted and stored at ≤-20°C
      Detailed protocolThe Mouse VEGF ELISA Kit is provided with removable well strips so the assay can be carried out on separate occasions. Since conditions may vary, a standard curve MUST be determined each time the assay is performed. Standards, controls and test samples should be assayed in duplicate. Disposable pipette tips and reagent troughs should be used for all transfers to avoid cross-contamination of reagents or samples. Bring all reagents to room temperature before use.
      1. Prepare all samples, controls, standards and reagents as described in the previous sections.
      2. Remove the appropriate number of wells from the foil pouch and place them into the empty well holder. Return any unused wells to the foil pouch, refold, seal and store at 4°C. Unused wells may be stored at 4°C for up to 1 month.
      3. Pipette 50 µl of Assay Diluent RD1N to each well.
      4. Add 50 µl of samples (standards, control and test sample*) into the appropriate wells on top of the Assay Diluent RD1N. Note: use clean pipette tips for each sample. Mix the sample/Assay Diluent RD1N mixture by gently tapping the plate frame for 1 min.
      5. Following mixing, cover the plate with the plate sealer provided and incubate the plate at room temperature for 2 h.
      6. Wash wells 5 times with 1X Wash Buffer making sure each well is filled completely. Each well is washed by filling with 400 µl of 1X Wash Buffer (using a multichannel pipette, automatic plate washer or wash bottle). It is essential to completely remove the Wash Buffer after each step and to ensure that the wells do not contain any buffer after the last wash. This can be achieved by inverting the plate and tapping it on paper towels.
      7. Add 100 µl of Mouse VEGF Conjugate solution for each of the samples, standards and control wells. Cover the plate with the plate sealer provided and incubate the plate at room temperature for 2 h.
      8. Wash wells 5 times with 1X Wash Buffer making sure each well is filled completely. Follow procedure as outlined in Step 6.
      9. Pipette 100 µl of Substrate Solution to each well and incubate in the dark at room temperature for 30 min.
      10. Add 100 µl of Stop Solution to each well in the same order as the previously added Substrate Solution. Mix the Substrate/Stop Solution mixture by gently tapping the plate frame.
      11. Measure absorbance in each well using a spectrophotometric plate reader at dual wavelengths of 450/540 nm (570 nm can be used as an alternative reference wavelength). If wavelength reference is not available read the plate at 540 or 570 nm and subtract this value from that obtained at 450 nm. Wells must be read within 30 min of adding the Stop Solution.

      Note: Samples that give values higher than the highest standard should be diluted with Calibrator Diluent and re-evaluated in the assay.
      Note: All mouse serum samples should firstly be diluted 1:5 in Calibrator Diluent RD5T as outlined in Sample Preparation.
      Calculations1. Average the duplicate absorbance values for the control, each standard (including the zero) and all samples. 2. On graph paper, plot the mean absorbance values for each of the standards on the Y axis, versus the concentration of each standard (pg/ml) on the X axis. 3. Determine the concentration of unknowns by interpolation from the standard curve. There are a variety of microplate reader software packages available (Softmax, Molecular Devices Corporation, Menlo Park, CA; KinetiCalc, BioTek Instruments, Inc. Winooski, VT) for analysis of microplate data, which simplify this process. 4. For samples which have been diluted (such as mouse serum samples), the mouse VEGF protein concentration must be multiplied by the dilution factor (i.e. since mouse serum samples are diluted five fold, then the mouse VEGF protein concentration value obtained from the standard curve must be multiplied by five).
      Assay characteristics and examplesCalibration: The mouse VEGF ELISA Kit is calibrated against an Sf 21-expressed, recombinant mouse VEGF protein. The recombinant protein is a disulfide linked homodimeric glycosylated molecule containing two 164 amino acid subunits with a total molecular weight of 42 kDa as demonstrated by non-reducing SDS-PAGE.

      Expected Values:

      I. Mouse Serum:
      Forty different mouse serum samples were evaluated for serum mouse VEGF protein levels using the Mouse VEGF ELISA. All samples demonstrated VEGF protein levels less than the lowest mouse VEGF standard of 7.8 pg/ml.

      II. Cell Culture Medium:
      Mouse macrophage cells (P388D1; 1x10⁵ cells/ml) were grown for 4 days in RPMI medium containing 5% FBS. The culture supernatant was assayed for mouse VEGF and contained 7.6 ng/ml.

      Mouse lung conditioned medium (1 lung; 1-2 mm pieces in 10 ml medium; 5 day culture) was assayed and found to contain 4.66 ng/ml mouse VEGF.

      Mouse heart conditioned medium (1 heart; 1-2 mm pieces in 10 ml medium; 5 day culture) was assayed and found to contain 378 pg/ml mouse VEGF.
      Standard curveA typical standard curve is shown below. This particular curve is for demonstration only and should not be used for data extrapolation purposes. Data should be extrapolated from standard curves run within, and in conjunction with the test samples. A standard curve should be run in duplicate each time an assay is performed.

      Figure 1: Standard Curve
      Sensitivity3.0 pg/ml
      Sensitivity NotesThe minimum dose of mouse VEGF protein detectable using this assay is ~3 pg/ml. This value is calculated by adding two standard deviations to the mean optical density of 20 zero standard replicates and calculating the corresponding concentration.
      Assay Range7.8-500 pg/ml
      Precision

      Table 1: Inter-Assay Precision

      This reflects the precision between assays. Samples containing three different concentrations of mouse VEGF were analyzed in 20 separate assays.


      Table 2: Intra-Assay Precision

      This reflects the precision within an assay. Samples containing three different concentrations of mouse VEGF were analyzed 20 times each, in a single assay.
      RecoveryThe average % recovery of mouse VEGF protein, spiked to three levels throughout the range of the assay in various matrices was evaluated.

      Table 3: Recovery
      LinearityThe assay linearity was assessed using five or more samples containing and/or spiked with various concentrations of mouse VEGF diluted with Calibrator Diluent. The measured mouse VEGF concentrations at each dilution within the range of the assay were within 20% of the expected values. Some typical are shown below:

      Table 4: Linearity

      *Note: All mouse serum samples should firstly be diluted 1:5 in Calibrator Diluent RD5T as outlined in Sample Preparation.
      SpecificityThe Mouse VEGF ELISA Kit detects both natural and recombinant mouse VEGF. No significant cross-reactivity or interference was observed with the following factors at a concentration of 50 ng/ml:

      Recombinant mouse: IL-1α, 1β, 2-10, 10 sR, 12, 13; C-10; G-CSF; GM-CSF; IFN-γ; KC; JE/MCP-1; LIF; M-CSF; MIP-1α, 1β, 2; SCF; TNF-α; Tpo.
      Recombinant human: FIT-3 Ligand; PIGF; PDGF-AA, AB.
      Other: hPDGF

      Note: Some cross-reactivity was observed with rhVEGF¹⁶⁵ (0.2% cross-reactivity) and rhVEGF¹⁶⁵/PIGF (0.05% cross-reactivity).
      Protocol Summary

      Figure 2: Protocol Summary
      Registered TrademarksCalbiochem® is a registered trademark of EMD Chemicals, Inc.
      Interactive Pathways™ is a trademark of EMD Chemicals, Inc.