Phone:
+1 877 302 8632
Fax:
+1 888 205 9894 (Toll-free)
E-Mail:
orders@antibodies-online.com

FLT1 ELISA Kit

FLT1 Reactivity: Human Colorimetric Sandwich ELISA 0.312-20 ng/mL Cell Culture Supernatant, Plasma, Serum
Catalog No. ABIN456775
  • Target See all FLT1 ELISA Kits
    FLT1 (Fms-Related tyrosine Kinase 1 (VEGFR1) (FLT1))
    Reactivity
    • 6
    • 6
    • 4
    • 2
    • 2
    • 2
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    Human
    Detection Method
    Colorimetric
    Method Type
    Sandwich ELISA
    Detection Range
    0.312-20 ng/mL
    Minimum Detection Limit
    0.312 ng/mL
    Application
    ELISA
    Purpose
    This immunoassay kit allows for the specific measurement of human Vascular endothelial growth factor receptor-1,VEGFR-1 concentrations in cell culture supernates, serum, and plasma.
    Sample Type
    Cell Culture Supernatant, Serum, Plasma
    Analytical Method
    Quantitative
    Specificity
    This assay recognizes recombinant and natural human VEGFR-1.
    Cross-Reactivity (Details)
    No significant cross-reactivity or interference was observed.
    Characteristics
    Homo sapiens,Human,Vascular endothelial growth factor receptor 1,VEGFR-1,Fms-like tyrosine kinase 1,FLT-1,Tyrosine-protein kinase FRT,Tyrosine-protein kinase receptor FLT,Vascular permeability factor
    Components
    Reagent (Quantity):
    • Assay plate (1),
    • Standard (2),
    • Sample Diluent (1×20 mL),
    • Assay Diluent A (1×10 mL),
    • Assay Diluent B (1×10 mL),
    • Detection Reagent A (1×120 μL),
    • Detection Reagent B (1×120 μL),
    • Wash Buffer(25 x concentrate) (1×30 mL),
    • Substrate (1×10 mL),
    • 2 Stop Solution (1×10 mL),
    • Plate sealer for 96 wells (5),
    • Instruction (1)
    Material not included
    Microplate reader. Pipettes and pipette tips. EP tube Deionized or distilled water.
    Featured
    Discover our best selling FLT1 ELISA Kit
    Top Product
    Discover our top product FLT1 ELISA Kit
  • Sample Volume
    100 μL
    Plate
    Pre-coated
    Protocol
    This assay employs the quantitative sandwich enzyme immunoassay technique. A monoclonal antibody specific for VEGFR-1 has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any VEGFR-1 present is bound by the immobilized antibody. An enzyme-linked polyclonal antibody specific for VEGFR-1 is added to the wells. Following a wash 2 to remove any unbound antibody-enzyme reagent, a substrate solution is added to the wells and color develops in proportion to the amount of VEGFR-1 bound in the initial step. The color development is stopped and the intensity of the color is measured.
    Reagent Preparation

    Bring all reagents to room temperature before use. Wash Buffer - If crystals have formed in the concentrate, warm to room temperature and mix gently until the crystals have completely dissolved. Dilute 30 mL of Wash Buffer Concentrate into deionized or distilled water to prepare 750 mL of Wash Buffer. Standard - Reconstitute the Standard with 1.0 mL of Sample Diluent. This reconstitution produces a stock solution. Allow the standard to sit for a minimum of 15 minutes with gentle agitation prior to making serial dilutions (Making serial dilution in the wells directly is not permitted). The undiluted standard serves as the high standard. The Sample Diluent serves as the zero standard (0 ng/ml).

    Sample Collection
    Cell culture supernates - Remove particulates by centrifugation and assay immediately or aliquot and store samples at ≤ -20 °C. Avoid repeated freeze-thaw cycles. Serum - Use a serum separator tube (SST) and allow samples to clot for 30 minutes before centrifugation for 15 minutes at approximately 1000 x g. Remove serum and assay immediately or aliquot and store samples at -20 °C. Plasma - Collect plasma using EDTA or heparin as an anticoagulant. Centrifuge samples for 15 minutes at 1000 x g at 2 - 8 °C within 30 minutes of collection. Store samples at ≤ -20 °C. Avoid repeated freeze-thaw cycles.
    Assay Procedure

    Allow all reagents to reach room temperature (Please do not dissolve the reagents at 37 °C directly.). All the reagents should be mixed thoroughly by gently swirling before pipetting. Avoid foaming. Keep appropriate numbers of strips for 1 experiment and remove extra strips from microtiter plate. Removed strips should be resealed and stored at 4 °C until the kits expiry date. Prepare all reagents, working standards and samples as directed in the previous sections. Please predict the concentration before assaying. If values for these are not within the range of the standard curve, users must determine the optimal sample dilutions for their particular experiments.
    1. Add 100 μL of Standard, Blank, or Sample per well. Cover with the Plate sealer. Incubate for 2 hours at 37 °C .
    2. Remove the liquid of each well, don ’ t wash.
    3. Add 100 μL of Detection Reagent A working solution to each well. Cover with the Plate sealer. Incubate for 1 hour at 37 °C . Detection Reagent A working solution may appear cloudy. Warm to room temperature and mix gently until solution appears uniform.
    4. Aspirate each well and wash, repeating the process three times for a total of three washes. Wash by filling each well with Wash Buffer (approximately 400 μL) using a squirt bottle, multi-channel pipette, manifold dispenser or autowasher. Complete removal of liquid at each step is essential to good performance. After the last wash, remove any remaining Wash Buffer by aspirating or decanting. Invert the plate and blot it against clean paper towels.
    5. Add 100 μL of Detection Reagent B working solution to each well. Cover with a new Plate sealer. Incubate for 1 hours at 37 °C .
    6. Repeat the aspiration/wash as in step 4.
    7. Add 90 μL of Substrate Solution to each well. Cover with a new Plate sealer. Incubate within 30 minutes at 37 °C . Protect from light.
    8. Add 50 μL of Stop Solution to each well. If color change does not appear uniform, gently tap the plate to ensure thorough mixing.
    9. Determine the optical density of each well at once, using a microplate reader set to 450 nm.
    Important Note:
    1. Absorbance is a function of the incubation time. Therefore, prior to starting the assay it is recommended that all reagents should be freshly prepared prior to use and all required strip-wells are secured in the microtiter frame. This will ensure equal elapsed time for each pipetting step, without interruption.
    2. Please carefully reconstitute Standards or working Detection Reagent A and B according to the instruction, and avoid foaming and mix gently until the crystals have completely dissolved. The reconstituted Standards can be used only once. This assay requires pipetting of small volumes. To minimize imprecision caused by pipetting, ensure that pipettors are calibrated. It is recommended to suck more than 10 μ l for once pipetting.
    3. To ensure accurate results, proper adhesion of plate sealers during incubation steps is necessary. Do not allow wells to sit uncovered for extended periods between incubation steps. Once reagents have been added to the well strips, DO NOT let the 5 strips DRY at any time during the assay.
    4. For each step in the procedure, total dispensing time for addition of reagents to the assay plate should not exceed 10 minutes.
    5. To avoid cross-contamination, change pipette tips between additions of each standard level, between sample additions, and between reagent additions. Also, use separate reservoirs for each reagent.
    6. The wash procedure is critical. Insufficient washing will result in poor precision and falsely elevated absorbance readings.
    7. Duplication of all standards and specimens, although not required, is recommended.
    8. Substrate Solution is easily contaminated. Please protect it from light.

    Calculation of Results

    Average the duplicate readings for each standard, control, and sample and subtract the average zero standard optical density. Create a standard curve by reducing the data using computer software capable of generating a four parameter logistic (4-PL) curve-fit. As an alternative, construct a standard curve by plotting the mean absorbance for each standard on the x-axis against the concentration on the y-axis and draw a best fit curve through the points on the graph. The data may be linearized by plotting the log of the SAA concentrations versus the log of the O.D. and the best fit line can be determined by regression analysis. It is recommended to use some related software to do this calculation, such as curve expert 13.0. This procedure will produce an adequate but less precise fit of the data. If samples have been diluted, the concentration read from the standard curve must be multiplied by the dilution factor.

    Restrictions
    For Research Use only
  • Handling Advice
    1. The kit should not be used beyond the expiration date on the kit label.
    2. Do not mix or substitute reagents with those from other lots or sources.
    3. If samples generate values higher than the highest standard, further dilute the samples with the Assay Diluent and repeat the assay. Any variation in standard diluent, operator, pipetting technique, washing technique,incubation time or temperature, and kit age can cause variation in binding.
    4. This assay is designed to eliminate interference by soluble receptors, ligands, binding proteins, and other factors present in biological samples. Until all factors have been tested in the Immunoassay, the possibility of interference cannot be excluded.
    Storage
    4 °C/-20 °C
    Storage Comment
    The Assay Plate, Standard, Detection Reagent A and Detection Reagent B should be stored at -20°C upon being received. After receiving the kit , Substrate should be always stored at 4°C.
  • Li, Liu, Bin, Wang, Chen, Xiu, Pei, Lai, Chen, Fan, Xie, Tao, Wu: "Mutant hypoxia inducible factor-1? improves angiogenesis and tissue perfusion in ischemic rabbit skeletal muscle." in: Microvascular research, Vol. 81, Issue 1, pp. 26-33, (2011) (PubMed).

  • Target See all FLT1 ELISA Kits
    FLT1 (Fms-Related tyrosine Kinase 1 (VEGFR1) (FLT1))
    Alternative Name
    FLT1 (FLT1 Products)
    Synonyms
    FLT ELISA Kit, FLT-1 ELISA Kit, VEGFR-1 ELISA Kit, VEGFR1 ELISA Kit, FLT1 ELISA Kit, flt1b ELISA Kit, sFlt1 ELISA Kit, AI323757 ELISA Kit, Flt-1 ELISA Kit, fms related tyrosine kinase 1 ELISA Kit, FMS-related tyrosine kinase 1 ELISA Kit, vascular endothelial growth factor receptor 1 ELISA Kit, fms-related tyrosine kinase 1 (vascular endothelial growth factor/vascular permeability factor receptor) ELISA Kit, FMS-like tyrosine kinase 1 ELISA Kit, vascular endothelial growth factor receptor-1 ELISA Kit, FLT1 ELISA Kit, Flt1 ELISA Kit, CpipJ_CPIJ012087 ELISA Kit, flt1 ELISA Kit, FLT-1 ELISA Kit
    Background
    Vascular endothelial growth factor receptor-1 (VEGF R1, also known as Flt-1) was originally discovered through the screening of a human placental cDNA library (1). It is a receptor tyrosine kinase (RTK) specific for the angiogenic factors VEGF (VEGF-A), PlGF, and VEGF-B. VEGF R1 is expressed in two forms via alternate splicing at the pre-mRNA level: a full-length, membrane bound receptor capable of transducing signal, and a truncated, soluble receptor (sVEGF R1) capable of sequestering ligand or dimerizing with full-length receptor and preventing signal transduction. Although VEGF R1 null mutations are lethal, deletions of the kinase domain are not, suggesting that the soluble form, or at least the extracellular domain, is all that is necessary for normal vascular development. Experimental and clinical administrations of sVEGF R1 have been employed successfully in the prevention of neovasculogenesis and tumor growth. The serum/plasma level of sVEGF R1 may vary by pathology, and may have prognostic value. It has been suggested that sVEGF R1 levels should be presented alongside VEGF levels in experimental and pathological conditions. For a review of VEGF R1. The human VEGF R1 gene produces two major transcripts of 3.0 and 2.4 kb, corresponding to the full-length receptor and soluble receptor, respectively. Full length VEGF R1 is an approximately 180 kDa glycoprotein featuring seven extracellular immunoglobulin (Ig)-like domains, a membrane-spanning region, and an intracellular tyrosine kinase domain containing a kinase insert sequence. The truncated sVEGF R1 consists of only the first six extracellular Ig-like domains. Ligand binding takes place within the first three N-terminal Ig-like domains while the fourth Ig-like domain is responsible for receptor dimerization, which is a prerequisite for activation through trans-phosphorylation. In addition to homodimers, VEGF R1 can form active heterodimers with VEGF R2. The soluble form of VEGF R1 forms inactive heterodimers with VEGF R2.
    Pathways
    RTK Signaling, Signaling Events mediated by VEGFR1 and VEGFR2, VEGFR1 Specific Signals
You are here:
Support