Multiplex Assay Kit for Hypoxia Inducible Factor 1 Alpha (HIF1a) ,etc. by FLIA (Flow Luminescence Immunoassay) Oryctolagus cuniculus (Rabbit) Multiplex ELISA

HIF1-A; MOP1; PASD8; Basic Helix-Loop-Helix Transcription Factor; ARNT-interacting protein; Basic-helix-loop-helix-PAS protein MOP1; Class E basic helix-loop-helix protein 78

(Note: Up to 8-plex in one testing reaction)

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  • Multiplex Assay Kit for Hypoxia Inducible Factor 1 Alpha (HIF1a) ,etc. by FLIA (Flow Luminescence Immunoassay) Packages (Simulation)
  • Multiplex Assay Kit for Hypoxia Inducible Factor 1 Alpha (HIF1a) ,etc. by FLIA (Flow Luminescence Immunoassay) Packages (Simulation)
  • Multiplex Assay Kit for Hypoxia Inducible Factor 1 Alpha (HIF1a) ,etc. by FLIA (Flow Luminescence Immunoassay) Results demonstration
  • LMA798Rb.jpg Typical Standard Curve
  • Certificate ISO9001: 2008, ISO13485: 2003 Registered

Recovery

Matrices listed below were spiked with certain level of recombinant Hypoxia Inducible Factor 1 Alpha (HIF1a) ,etc. by FLIA (Flow Luminescence Immunoassay) and the recovery rates were calculated by comparing the measured value to the expected amount of Hypoxia Inducible Factor 1 Alpha (HIF1a) ,etc. by FLIA (Flow Luminescence Immunoassay) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 92-99 96
EDTA plasma(n=5) 83-101 86
heparin plasma(n=5) 84-92 87
sodium citrate plasma(n=5) 93-105 102

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Hypoxia Inducible Factor 1 Alpha (HIF1a) ,etc. by FLIA (Flow Luminescence Immunoassay) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Hypoxia Inducible Factor 1 Alpha (HIF1a) ,etc. by FLIA (Flow Luminescence Immunoassay) were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: CV<10%
Inter-Assay: CV<12%

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Hypoxia Inducible Factor 1 Alpha (HIF1a) ,etc. by FLIA (Flow Luminescence Immunoassay) and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.

Sample 1:2 1:4 1:8 1:16
serum(n=5) 91-101% 90-101% 91-105% 80-89%
EDTA plasma(n=5) 86-94% 97-105% 99-105% 91-98%
heparin plasma(n=5) 80-101% 83-90% 94-101% 98-105%
sodium citrate plasma(n=5) 95-103% 99-105% 79-99% 83-92%

Stability

The stability of kit is determined by the loss rate of activity. The loss rate of this kit is less than 5% within the expiration date under appropriate storage condition.
To minimize extra influence on the performance, operation procedures and lab conditions, especially room temperature, air humidity, incubator temperature should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same operator from the beginning to the end.

Reagents and materials provided

Reagents Quantity Reagents Quantity
96-well plate 1 Plate sealer for 96 wells 4
Pre-Mixed Standard 2 Standard Diluent 1×20mL
Pre-Mixed Magnetic beads (22#:HIF1a) 1 Analysis buffer 1×20mL
Pre-Mixed Detection Reagent A 1×120μL Assay Diluent A 1×12mL
Detection Reagent B (PE-SA) 1×120μL Assay Diluent B 1×12mL
Sheath Fluid 1×10mL Wash Buffer (30 × concentrate) 1×20mL
Instruction manual 1

Assay procedure summary

1. Preparation of standards, reagents and samples before the experiment;
2. Add 100μL standard or sample to each well,
    add 10μL magnetic beads, and incubate 90min at 37°C on shaker;
3. Remove liquid on magnetic frame, add 100μL prepared Detection Reagent A. Incubate 60min at 37°C on shaker;
4. Wash plate on magnetic frame for three times;
5. Add 100μL prepared Detection Reagent B, and incubate 30 min at 37°C on shaker;
6. Wash plate on magnetic frame for three times;
7. Add 100μL sheath solution, swirl for 2 minutes, read on the machine.

Multiplex Assay Kit for Hypoxia Inducible Factor 1 Alpha (HIF1a) ,etc. by FLIA (Flow Luminescence Immunoassay)

Test principle

Analyte-specific antibodies are pre-coated onto color-coded microparticles. Microparticles, standards, and samples are pipetted into wells and the immobilized antibodies bind the analytes of interest. After washing away any unbound substances, a biotinylated antibody cocktail specific to the analytes of interest is added to each well. Following a wash to remove any unbound biotinylated antibody, Streptavidin-Phycoerythrin conjugate (Streptavidin-PE), which binds to the biotinylated detection antibodies, is added to each well. A final wash removes unbound Streptavidin-PE and the microparticles are resuspended in buffer and read using the Luminex or Bio-Plex analyzer.The MFI developed is proportional to the concentration of analytes of interest in the sample.

Giveaways

Citations

  • Quercetin greatly improved therapeutic index of doxorubicin against 4T1 breast cancer by its opposing effects on HIF-1α in tumor and normal cells.PubMed: 19466611
  • Baicalin suppresses lung carcinoma and lung metastasis by SOD mimic and HIF-1α inhibitionScienceDirect: S0014299909011352
  • Lactate Dehydrogenase 5 Expression in Non-Hodgkin Lymphoma Is Associated with the Induced Hypoxia Regulated Protein and Poor PrognosisPubMed: PMC3781153
  • Selective inhibition of PKCβ2 preserves cardiac function after myocardial infarction and is associated with improved angiogenesis of ischemic myocardium in diabetic ratsPubmed: 24002353
  • Modulation of arginine and asymmetric dimethylarginine concentrations in liver and plasma by exogenous hydrogen sulfide in LPS-induced endotoxemiaPubmed: 24289078
  • Method for preventing or treating a disease, disorder or condition induced by retina ischemiaUS8569372
  • Effects of adrenomedullin and vascular endothelial growth factor on ischemia/reperfusion injury in skeletal muscle in rats.Pubmed: 23816242
  • HIF-1α Is Essential for Effective PMN Bacterial Killing, Antimicrobial Peptide Production and Apoptosis in Pseudomonas aeruginosa KeratitisPlos: Source
  • Telomeres are elongated in rats exposed to moderate altitudePubmed:Pmc4088304
  • Resveratrol abrogates the effects of hypoxia on cell proliferation, invasion and EMT in osteosarcoma cells through downregulation of the HIF-1α protein.Pubmed:25384583
  • The enhancement of gene silencing efficiency with chitosan-coated liposome formulations of siRNAs targeting HIF-1α and VEGFPubmed:25445537
  • Effect of combined treatment with rosuvastatin and protein kinase Cβ2 inhibitor on angiogenesis following myocardial infarction in diabetic ratsPubmed:25524396
  • Maternal omega-3 fatty acid supplementation to a vitamin B 12 deficient diet normalizes angiogenic markers in the pup brain at birthPubMed: 25889224
  • Determination of the Preferred Conditions for the Isolated Perfusion of Porcine KidneysPubMed: 25341590
  • Oxygen-induced cell migration and on-line monitoring biomarkers modulation of cervical cancers on a microfluidic systemPubMed: 25905434
  • Metabolomic Analysis of Biochemical Changes in the Plasma of High-Fat Diet and Streptozotocin-Induced Diabetic Rats after Treatment with Isoflavones Extract of Radix Puerariaepmc:PMC4794592
  • Roles of Copper in Hepatocarcinogenesis via the Activation of Hypoxia-Inducible Factor-1αPubmed:27121973
  • Использование гипоксического прекондиционирования для подготовки аутодермотрансплантата у больных со скомпрометированной микроциркуляцией91
  • Roles of Copper in Hepatocarcinogenesis via the Activation of Hypoxia-Inducible Factor-1α.pubmed:27121973
  • Expression of hypoxia-inducible factor-1α in synovial fluid and articular cartilage is associated with disease severity in knee osteoarthritisetm:13
  • Correlating oxidative stress-related factors with bone metabolic markers in older adult male patients exhibiting degenerative osteoporosis in the high-altitude hypoxic area of China: study protocol for a non-randomized controlled trialissn:2542-3975
  • R132H Mutation in IDH1 Gene is Associated with Increased Tumor HIF1-Alpha and Serum VEGF Levels in Primary Glioblastoma Multiforme pubmed:28667042
  • Is there any relationship between adipocytokines and angiogenesis factors to address endothelial dysfunction and platelet aggregation in untreated patients with preeclampsia?10.1007/s00404-017-4461-2
  • Expression of hypoxia-inducible factor-1α in synovial fluid and articular cartilage is associated with disease severity in kneepubmed:28123469
  • Study of acute hypoxia markers in healthy subjects: Utility in post-crash investigation10.1016/j.mjafi.2017.04.003
  • Telomere elongation protects heart and lung tissue cells from fatal damage in rats exposed to severe hypoxiaPubmed:29454386
  • Signal Mechanism of the Protective Effect of Combined Preconditioning by Amtizole and Moderate HypoxiaPubmed:29308565
  • Determination of hypoxia-inducible factor-1 by using a ratiometric colorimetric test based on click-mediated growth of gold nanoparticlesPubmed: 30209641
  • Expression of Hif-1α, Nf-κb, and Vegf Genes in the Liver and Blood Serum Levels of HIF-1α, Erythropoietin, VEGF, TGF-β, 8-Isoprostane, and Corticosterone in Wistar Rats with High and Low Resistance to Hypoxia. Corticosterone in …Pubmed: 30353332
  • OKN-007 Increases temozolomide (TMZ) Sensitivity and Suppresses TMZ-Resistant Glioblastoma (GBM) Tumor GrowthPubmed: 30468988
  • Plasma Cellular Hypoxia, Mitochondrial Dysfunction, Disease Risk and Prognostic Factors in Type 2 Diabetic Patients in Saudi Arabia.
  • Age-Specific Features of Hypoxia Tolerance and Intensity of Lipopolysaccharide-Induced Systemic Inflammatory Response in Wistar RatsPubmed: 30903510
  • The role of amniotic fluid exosome, hypoxia inducible fator-1α, and inflammatory cytokine in cervical insufficiency
  • Vitexin alleviates interleukin‐1β induced inflammatory responses in chondrocytes from osteoarthritis patients: involvement of HIF‐1α pathwayPubmed: 31055848
  • A disintegrin‐like and metalloproteinase with thrombospondin‐1 (ADAMTS‐1) levels in gingival crevicular fluid correlate with vascular endothelial growth factor‐A …Pubmed: 31020669
  • Hypoxia Preconditioned Adipose Derived Endothelial Progenitor Cells Promote Bladder AugmentationPubmed: 31238789
  • ПОТЕНЦИРОВАНИЕ АНТИГИПОКСАНТАМИ ЭФФЕКТА ГИПОКСИЧЕСКОГО ПРЕКОНДИЦИОНИРОВАНИЯ
  • Maternal omega-3 fatty acids and vitamin E improve placental angiogenesis in late-onset but not early-onset preeclampsiaPubmed: 31420792
  • Regulation of MicroRNA-155 and Its Related Genes Expression by Inositol Hexaphosphate in Colon Cancer CellsPubmed: 31744065
  • Short-wave enhances mesenchymal stem cell recruitment in fracture healing by increasing HIF-1 in callusPubmed: 32894200
  • PLCε regulates metabolism and metastasis signaling via HIF‐1α/MEK/ERK pathway in prostate cancerPubmed: 32383180
  • Гипоксия-зависимый транскрипционный контроль активности деструктивных изменений пародонта воспалительного и злокачественного генеза
  • Prenatal vitamin D supplementation reduces blood pressure and improves placental angiogenesis in an animal model of preeclampsia33237074
  • Lactucaxanthin protects retinal pigment epithelium from hyperglycemia-regulated hypoxia/ER stress/VEGF pathway mediated angiogenesis in ARPE-19 cell and rat ¡­33705802
  • Astaxanthin mediated regulation of VEGF through HIF1¦Á and XBP1 signaling pathway: An insight from ARPE-19 cell and streptozotocin mediated diabetic rat model33789142
  • Serum hypoxia-inducible factor-1¦Á and uterine artery Doppler ultrasound during the first trimester for prediction of preeclampsia33758274
  • Umbilical cord-derived mesenchymal stem cell conditioned medium reverses neuronal oxidative injury by inhibition of TRPM2 activation and the JNK signaling …Pubmed:35585377

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