Human HIF-1α(Hypoxia Inducible Factor 1 Alpha) ELISA Kit

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    • sandwich-Ab-ELISA-Elabscience
    • sandwich-Ab-ELISA-Elabscience
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    • sandwich-Ab-ELISA-Elabscience
    • sandwich-Ab-ELISA-Elabscience
    • sandwich-Ab-ELISA-Elabscience
    • sandwich-Ab-ELISA-Elabscience
    • sandwich-Ab-ELISA-Elabscience
    • sandwich-Ab-ELISA-Elabscience

      Catalog number:E-EL-H1277

      Synonyms:HIF1A, HIF-1A, MOP1, PASD8, bHLHe78, basic helix-loop-helix transcription factor

      Size:
      • 96T
      • 24T
      Qty:
      - +
      Price: $495

      Reactivity: Human

      Detection Range: 0.16~10 ng/mL

      Sensitivity: 0.10 ng/mL

      Lead Time: 7~10 daysWelcome to order from local distributors.

      Add to cart Compare Bulk request

      Intended use

      This ELISA kit applies to the in vitro quantitative determination of Human HIF-1α concentrations in serum, plasma and other biological fluids.

      Test principle

      This ELISA kit uses the Sandwich-ELISA principle. The micro ELISA plate provided in this kit has been pre-coated with an antibody specific to Human HIF-1α. Standards or samples are added to the micro ELISA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Human HIF-1α and Avidin-Horseradish Peroxidase (HRP) conjugate are added successively to each micro plate well and incubated. Free components are washed away. The substrate solution is added to each well. Only those wells that contain Human HIF-1α, biotinylated detection antibody and Avidin-HRP conjugate will appear blue in color. The enzyme-substrate reaction is terminated by the addition of stop solution and the color turns yellow. The optical density (OD) is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The OD value is proportional to the concentration of Human HIF-1α. You can calculate the concentration of Human HIF-1α in the samples by comparing the OD of the samples to the standard curve.

      Assay type Sandwich
      Format 96T
      Assay time 3.5h
      Reactivity Human
      Detection Method Colormetric
      Detection Range 0.16—10 ng/mL
      Sensitivity 0.10 ng/mL
      Sample Volume Required Per Well 100μL
      Sample Type Serum, plasma and other biological fluids

      Specificity

      This kit recognizes Human HIF-1α in samples. No significant cross-reactivity or interference between Human HIF-1α and analogues was observed.

      Typical data

      As the OD values of the standard curve may vary according to the conditions of the actual assay performance (e.g. operator, pipetting technique, washing technique or temperature effects), the operator should establish a standard curve for each test. Typical standard curve and data is provided below for reference only.

      Concentration
      (ng/mL)
      O.D Average Corrected
      10 2.341
      2.373
      2.357 2.269
      5 1.557
      1.601
      1.579 1.491
      2.5 0.882
      0.874
      0.878 0.79
      1.25 0.461
      0.469
      0.465 0.377
      0.63 0.267
      0.245
      0.256 0.168
      0.31 0.188
      0.178
      0.183 0.095
      0.16 0.129
      0.149
      0.139 0.051
      0 0.086
      0.09
      0.088 --

      Precision

      Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human HIF-1α were tested 20 times on one plate, respectively.
      Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human HIF-1α were tested on 3 different plates, 20 replicates in each plate.

      Intra-assay Precision Inter-assay Precision
      Sample 1 2 3 1 2 3
      n 20 20 20 20 20 20
      Mean (ng/mL) 0.50 1.60 3.70 0.50 1.70 3.60
      Standard deviation 0.03 0.08 0.11 0.03 0.07 0.18
      C V (%) 6.00 5.00 2.97 6.00 4.12 5.00

      Recovery

      The recovery of Human HIF-1α spiked at three different levels in samples throughout the range of the assay was evaluated in various matrices.

      Sample Type Range (%) Average Recovery (%)
      Serum (n=5) 90-101 96
      EDTA plasma (n=5) 88-102 93
      Cell culture media (n=5) 92-107 98

      Linearity

      Samples were spiked with high concentrations of Human HIF-1α and diluted with Reference Standard & Sample Diluent to produce samples with values within the range of the assay.

      Serum (n=5) EDTA plasma (n=5) Cell culture media (n=5)
      1:2 Range (%) 95-110 91-106 89-102
      Average (%) 101 99 96
      1:4 Range (%) 95-106 86-99 92-105
      Average (%) 101 92 97
      1:8 Range (%) 98-115 88-99 96-109
      Average (%) 105 93 103
      1:16 Range (%) 93-109 87-101 96-113
      Average (%) 101 93 104

      Kit components & Storage

      An unopened kit can be stored at 2-8°C for 1 month. If the kit is not used within 1 month, store the items separately according to the following conditions once the kit is received.

      Item Specifications Storage
      Micro ELISA Plate(Dismountable) 8 wells ×12 strips -20°C, 6 months
      Reference Standard 2 vials
      Concentrated Biotinylated Detection Ab (100×) 1 vial, 120 μL
      Concentrated HRP Conjugate (100×) 1 vial, 120 μL -20°C(shading light), 6 months
      Reference Standard & Sample Diluent 1 vial, 20 mL 2-8°C, 6 months
      Biotinylated Detection Ab Diluent 1 vial, 14 mL
      HRP Conjugate Diluent 1 vial, 14 mL
      Concentrated Wash Buffer (25×) 1 vial, 30 mL
      Substrate Reagent 1 vial, 10 mL 2-8°C(shading light)
      Stop Solution 1 vial, 10 mL 2-8°C
      Plate Sealer 5 pieces
      Product Description 1 copy
      Certificate of Analysis 1 copy

      Note: All reagent bottle caps must be tightened to prevent evaporation and microbial pollution.
      The volume of reagents in partial shipments is a little more than the volume marked on the label, please use accurate measuring equipment instead of directly pouring into the vial(s).

      Other supplies required

      • Microplate reader with 450 nm wavelength filter
      • High-precision transfer pipette, EP tubes and disposable pipette tips
      • Incubator capable of maintaining 37°C
      • Deionized or distilled water
      • Absorbent paper
      • Loading slot for Wash Buffer

      Assay Procedure

      elisa assay procedure 1

      1. Add 100μL standard or sample to the wells. Incubate for 90 min at 37°C

      elisa assay procedure 2

      2. Discard the liquid, immediately add 100μL Biotinylated Detection Ab working solution to each well. Incubate for 60 min at 37°C

      elisa assay procedure 3

      3. Aspirate and wash the plate for 3 times

      elisa assay procedure 4

      4. Add 100μL HRP conjugate working solution. Incubate for 30 min at 37°C. Aspirate and wash the plate for 5 times

      elisa assay procedure 5

      5. Add 90μL Substrate Reagent. Incubate for 15 min at 37°C

      elisa assay procedure 6

      6. Add 50μL Stop Solution

      elisa assay procedure 7

      7. Read the plate at 450nm immediately. Calculation of the results

      Citations

      1. Publication: Chandravanshi B, Bhonde R. Small Molecules Exert Anti-Apoptotic Effect And Reduce Oxidative Stressaugmenting Insulin Secretion In Stem Cells Engineered Isletsagainst Hypoxia[J]. European Journal of Pharmacology, 2016, 791: 424-432.
        Species: Human
        Sample Type: Culture supernatant
      2. Publication: Nath M, Chandra P, Halder N, et al. Involvement Of Renin-Angiotensin System In Retinopathy Of Prematurity - A Possible Target For Therapeutic Intervention[J]. Plos One, 2016.
        Species: Human
        Sample Type: Vitreous humour
      3. Publication: Chandravanshi B, Bhonde R R. Shielding Engineered Islets With Mesenchymal Stem Cells Enhance Survival Under Hypoxia[J]. Journal of Cellular Biochemistry, 2017.
        Species: Human
        Sample Type: Cell lysis
      4. Publication: Mathew S A, Chandravanshi B, Bhonde R. Hypoxia Primed Placental Mesenchymal Stem Cells For Wound Healing[J]. Life Sciences, 2017.
        Species: Human
        Sample Type: cell lysis
      5. Publication: Jalkanen J, Maksimow M, Jalkanen S, et al. Hypoxia-Induced Inflammation And Purinergic Signaling In Cross Clamping The Human Aorta[J]. Springerplus, 2016.
        Species: Human
        Sample Type: Serum
      6. Publication: Tan F, Zhu H, He X, et al. Role of TXNDC5 in tumorigenesis of colorectal cancer cells: In vivo and in vitro evidence[J]. International Journal of Molecular Medicine, 2018, 42(2): 935-945.
        Species: Human
        Sample Type: Tissue
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