HIF1A Kit ELISA

N° du produit ABIN1115458

Détail du produit HIF1A Kit ELISA

Antigène: HIF1A (Hypoxia Inducible Factor 1, alpha Subunit (Basic Helix-Loop-Helix Transcription Factor) (HIF1A))

Reactivité: Humain

Méthode de détection: Colorimetric

Type de méthode: Sandwich ELISA

Gamme de detection: 62.5 pg/mL - 4000 pg/mL

Seuil minimal de détection: 62.5 pg/mL

Application: ELISA

Fonction: The kit is a sandwich enzyme immunoassay technique for the in vitro quantitative measurement in various sample types.

Type d'échantillon: Plasma, Serum

Analytical Method: Quantitative

Specificité: This kit recognizes natural and recombinantHumanHIF-1α. No significant cross-reactivity or interference between HumanHIF-1α and analogues was observed. Note: Limited by existing techniques, cross reaction may still exist, as it is impossible for us to complete the cross-reactivity detection between HumanHIF-1α and all the analogues.

Réactivité croisée (Details): No significant cross-reactivity or interference between human Hypoxia Inducible Factor 1 Alpha and analogues was observed. Note: Limited by current skills and knowledge, it is impossible for us to complete the cross-reactivity detection between Human bFGF and all the analogues, therefore, cross reaction may still exist.

Sensibilité: 37.5 pg/mL

Ingrédients:

• Pre-coated, ready to use 96-well strip plate, flat buttom
• Plate sealer for 96 wells
• Reference Standard
• Reference Standard & Sample Diluent
• Biotinylated Detection Antibody (100 x concentrate)
• HRP Conjugate (100 x concentrate)
• Biotinylated Detection Antibody Diluent
• HRP Conjugate Diluent
• Substrate Reagent
• Stop Solution
• Wash Buffer (25 x concentrate)
• Instruction manual

Information d'application

Indications d'application: ELISA Plate: The just opened ELISA Plate may appear water-like substance, which is normal and will not have any impact on the experiment results.
Add Sample: The interval of sample adding between the first well and the last well should not be too long, otherwise will cause different pre-incubation time, which will significantly affect the experiment’s accuracy and repeatability. For each step in the procedure, total dispensing time for addition of reagents or samples to the assay plate should not exceed 10 minutes. Parallel measure ment is recommended.
Incubation: To prevent evaporation and 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. Do not let the strips dry at any time during the assay. Strict compliance with the given incubation time and temperature.
Washing: The wash procedure is critical. Insufficient washing will result in poor precision and falsely elevated absorbance readings. Residual liquid in the reaction wells should be pat dry against absorbent paper in the washing process. But don’t put absorbent paper into reaction wells directly. Note that clear the residual liquid and fingerprint in the bottom before measurement, so as not to affect the micro-titer plate reader.
Reagent Preparation: As the volume of Detection Ab and HRP Conjugate is very small, liquid may adhere to the tube wall or tube cap when being transported. You better hand-throw it or centrifugal it for 1 minute at 1000rpm. Please pipette the solution for 4-5 times before pippeting. Please carefully reconstitute Standards, working solutions of Detection Ab and HRP Conjugate according to the instructions. To minimize imprecision caused by pipetting, ensure that pipettors are calibrated. It is recommended to suck more than 10μL for once pipetting. Do not reuse standard solution, working solution of Detection Ab and HRP Conjugate, which have been diluted. If you need to use standard repeatedly, you can divide the standard into small pack according to the amount of each assay, keep them at -20°C to -80°C and avoid repeated freezing and thawing.
Reaction Time Control: Please control reaction time strictly following this product description!
Substrate: Substrate Solution is easily contaminated. Please protect it from light.Stop Solution: As it is an acid solution, please pay attention to the protection of your eyes, hands, face and clothes when using this solution.
Mixing: You’d better use micro-oscillator at the lowest frequency, as sufficient and gentle mixing is particularly important to reaction result. If there is no micro-oscillator available, you can knock the ELISA plate frame gently with your finger before reaction.
Security: Please wear lab coats and latex gloves for protection. Especially detecting samples of blood or other body fluid, please perform following the national security columns of biological laboratories.
Do not use component from different batches of kit(washing buffer and stop solution can be an exception)
To avoid cross-contamination, change pipette tips between adding of each standard level, between sample adding, and between reagent adding. Also, use separate reservoirs for each reagent. Otherwise, the results will be inaccurate!

Commentaires:

Information on standard material:
The formulation of the standard is 0.01 M PBS. The standard contains additives (1 % BSA).

Information on reagents:
Reagents include 1 M SO₂. Azide, thimerosal, 2-mercaptoethanol (2-ME) or any other poisonous materials are not used.

Information on antibodies:
The provided antibodies and their host vary in different kits. All antibodies are affinity purified

The sensitivity of this assay, or Lower Limit of Detection (LLD) was defined as the lowest protein concentration that could be differentiated from zero. It was determined by adding two standard deviations to the mean optical density value of twenty zero standard replicates and calculating the corresponding concentration.

Volume d'échantillon: 100 µL

Plaque: Pre-coated

Protocole:

1. Add 100 µL standard or sample to each well. Incubate for 90 min at 37 °C.
2. Remove the liquid. Add 100 µL Biotinylated Detection Antibody. Incubate for 1 hour at 37 °C.
3. Aspirate and wash 3 times.
4. Add 100 µL HRP Conjugate. Incubate for 30 min at 37 °C.
5. Aspirate and wash 5 times.
6. Add 90 µL Substrate Reagent. Incubate for 15 min at 37 °C.
7. Add 50 µL Stop Solution. Read at 450 nm immediately.
8. Calculation of results.

Préparation des réactifs:

1. Bring all reagents to room temperature (18-25 °C) before use. If the kit will not be used up in one assay, please only take out the necessary strips and reagents for present experiment, and store the remaining strips and reagents at required condition.
2. Wash Buffer: Dilute 30 mL of Concentrated Wash Buffer with 720 mL of deionized or distilled water to prepare 750 mL of Wash Buffer. Note: if crystals have formed in the concentrate, warm it in a 40 °C water bath and mix it gently until the crystals have completely dissolved.
3. Standard working solution: Centrifuge the standard at 10,000xg for 1 min. Add 1.0 mL of Reference Standard &Sample Diluent, let it stand for 10 min and invert it gently several times. After it dissolves fully, mix it thoroughly with a pipette. This reconstitution produces a working solution of 4000pg/mL(or add 1.0 mL of Reference Standard &Sample Diluent, let it stand for 1-2 min and then mix it thoroughly with a vortex meter of low speed. Bubbles generated during vortex could be removed by centrifuging at a relatively low speed). Then make serial dilutions as needed. The recommended dilution gradient is as follows: 4000, 2000, 1000, 500, 250, 125, 62.5, 0pg/mL. Dilution method: Take 7 EP tubes, add 500μL of Reference Standard & Sample Diluent to each tube. Pipette 500μL of the 4000pg/mL working solution to the first tube and mix up to produce a 2000pg/mL working solution. Pipette 500μL of the solution from the former tube into the latter one according to this step. The illustration below is for reference. Note: the last tube is regarded as a blank. Don't pipette solution into it from the former tube.
4. Biotinylated Detection Antibody working solution: Calculate the required amount before the experiment (100 μL/well). In preparation, slightly more than calculated should be prepared. Centrifuge the Concentrated Biotinylated Detection Antibody at 800xg for 1 min, then dilute the 100x Concentrated Biotinylated Detection Antibody to 1x working solution with Biotinylated Detection Antibody Diluent(Concentrated Biotinylated Detection Antibody: Biotinylated Detection Antibody Diluent= 1: 99).
5. HRP Conjugate working solution: Calculate the required amount before the experiment (100 μL/well). In preparation, slightly more than calculated should be prepared. Centrifuge the Concentrated HRP Conjugate at 800xg for 1 min, then dilute -6- the 100x Concentrated HRP Conjugate to 1x working solution with HRP Conjugate Diluent(Concentrated HRP Conjugate: HRP Conjugate Diluent= 1: 99). 4000 2000 1000 500 250 125 62.5 0

Restrictions: For Research Use only

Stockage

Conseil sur la manipulation: All the reagents in the kit should be stored according to the labels on vials. Unused wells should be returned to the foil pouch with the desiccant pack and resealed along entire edge of zip-seal. Substrate Reagent shouldn't be kept at -20 °C (Check!). Exposure of reagents to strong light should be avoided in the process of incubation and storage. All the taps of reagents should be tightened to prevent evaporation and microbial contamination. If not to store reagents according to above suggestions, erroneous results may occur.

Stock: 4 °C/-20 °C

Stockage commentaire: The unopened kit can be stored at 4℃ for 1 month. If the kit is not used within 1 month, store the items separately according to the conditions since the kit is received.

Références pour HIF1A Kit ELISA (ABIN1115458)

Tan, Zhu, He, Yu, Zhang, Xu, Pei: "Role of TXNDC5 in tumorigenesis of colorectal cancer cells: In vivo and in vitro evidence." dans: International journal of molecular medicine, Vol. 42, Issue 2, pp. 935-945, (2018) (PubMed).

Mathew, Chandravanshi, Bhonde: "Hypoxia primed placental mesenchymal stem cells for wound healing." dans: Life sciences, Vol. 182, pp. 85-92, (2017) (PubMed).

Nath, Chandra, Halder, Singh, Deorari, Kumar, Azad, Velpandian: "Involvement of Renin-Angiotensin System in Retinopathy of Prematurity - A Possible Target for Therapeutic Intervention." dans: PLoS ONE, Vol. 11, Issue 12, pp. e0168809, (2017) (PubMed).

Chandravanshi, Bhonde: "Small molecules exert anti-apoptotic effect and reduce oxidative stress augmenting insulin secretion in stem cells engineered islets against hypoxia." dans: European journal of pharmacology, Vol. 791, pp. 424-432, (2016) (PubMed).

Jalkanen, Maksimow, Jalkanen, Hakovirta: "Hypoxia-induced inflammation and purinergic signaling in cross clamping the human aorta." dans: SpringerPlus, Vol. 5, pp. 2, (2016) (PubMed).

Détail du antigène

Antigène: HIF1A (Hypoxia Inducible Factor 1, alpha Subunit (Basic Helix-Loop-Helix Transcription Factor) (HIF1A))

Autre désignation: Hypoxia Inducible Factor 1 Alpha (HIF1A Produits)

Synonymes: hif1a Kit ELISA, AA959795 Kit ELISA, HIF1alpha Kit ELISA, MOP1 Kit ELISA, bHLHe78 Kit ELISA, HIF-1A Kit ELISA, HIF-1alpha Kit ELISA, HIF1 Kit ELISA, HIF1-ALPHA Kit ELISA, PASD8 Kit ELISA, HIF1-alpha Kit ELISA, hif-1a Kit ELISA, hypoxia inducible factor 1 alpha Kit ELISA, hypoxia inducible factor 1, alpha subunit Kit ELISA, hypoxia inducible factor 1 alpha subunit Kit ELISA, HIF (Hypoxia Inducible Factor) homologa Kit ELISA, hif-1a Kit ELISA, Hif1a Kit ELISA, HIF1A Kit ELISA, hif1a Kit ELISA, hif-1 Kit ELISA

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

Pathways: Positive Regulation of Peptide Hormone Secretion, Regulation of Hormone Metabolic Process, Regulation of Hormone Biosynthetic Process, Cellular Response to Molecule of Bacterial Origin, Carbohydrate Homeostasis, Transition Metal Ion Homeostasis, Tube Formation, Regulation of Carbohydrate Metabolic Process, Signaling Events mediated by VEGFR1 and VEGFR2, VEGFR1 Specific Signals, L'effet Warburg