PDPK1 ELISA Kit

Produktnummer ABIN6953358

Produktdetails PDPK1 ELISA Kit

Target: PDPK1 (3-phosphoinositide Dependent Protein Kinase-1 (PDPK1))

Reaktivität: Maus

Nachweismethode: Colorimetric

Methodentyp: Sandwich ELISA

Detektionsbereich: 0.15 ng/mL - 10 ng/mL

Untere Nachweisgrenze: 0.15 ng/mL

Applikation: ELISA

Verwendungszweck: The kit is a sandwich enzyme immunoassay for in vitro quantitative measurement of PDPK1 in mouse tissue homogenates, cell lysates.

Proben: Cell Lysate, Tissue Homogenate

Analytische Methode: Quantitative

Spezifität: This assay has high sensitivity and excellent specificity for detection of Phosphoinositide Dependent Protein Kinase 1 (PDPK1)

Sensitivität: 0.055 ng/mL

Bestandteile:

• Pre-coated, ready to use 96-well strip plate, flat buttom
• Plate sealer for 96 wells
• Reference Standard
• Standard Diluent
• Detection Reagent A
• Detection Reagent B
• Assay Diluent A
• Assay Diluent B
• Reagent Diluent (if Detection Reagent is lyophilized)
• TMB Substrate
• Stop Solution
• Wash Buffer (30 x concentrate)
• Instruction manual

Anwendungsinformationen

Kommentare:

Information on standard material:
The standard might be recombinant protein or natural protein, that will depend on the specific kit. Moreover, the expression system is E.coli or yeast or mammal cell. There is 0.05% proclin 300 in the standard as preservative.

Information on reagents:
The stop solution used in the kit is sulfuric acid with concentration of 1 mol/L. And the wash solution is TBS. The standard diluent contains 0.02 % sodium azide, assay diluent A and assay diluent B contain 0.01% sodium azide. Some kits can contain is BSA in them.

Information on antibodies:
The provided antibodies and their host vary in different kits.

Probenmenge: 100 μL

Testdauer: 3 h

Plattentyp: Pre-coated

Protokoll:

1. Prepare all reagents, samples and standards,
2. Add 100μL standard or sample to each well. Incubate 1 hours at 37 °C,
3. Aspirate and add 100μL prepared Detection Reagent A. Incubate 1 hour at 37 °C,
4. Aspirate and wash 3 times,
5. Add 100μL prepared Detection Reagent B. Incubate 30 minutes at 37 °C,
6. Aspirate and wash 5 times,
7. Add 90μL Substrate Solution. Incubate 10-20 minutes at 37 °C,
8. Add 50μL Stop Solution. Read at 450nm immediately.

Aufbereitung der Reagenzien:

1. Bring all kit components and samples to room temperature (18-25 °C) before use.
2. Standard - Reconstitute the Standard with 1.0 mL of Standard Diluent, keep for 10 minutes at room temperature, shake gently (not to foam). The concentration of the standard in the stock solution is 10 ng/mL. Prepare 7 tubes containing 0.5 mL Standard Diluent and produce a double dilution series. Mix each tube thoroughly before the next transfer. Set up 7 points of diluted standard such as 10 ng/mL, 5 ng/mL, 2.5 ng/mL, 1.25 ng/mL, 0.625 ng/mL, 0.312 ng/mL, 0.156 ng/mL, and the last microcentrifuge tube with Standard Diluent is the blank as 0 ng/mL.
3. Assay Diluent A and Assay Diluent B - Dilute 6 mL of Assay Diluent A or B Concentrate(2x) with 6 mL of deionized or distilled water to prepare 12 mL of Assay Diluent A or B. (In fact, more than 6 mL Assay Diluent A and Assay Diluent B are contained in the bottles. Therefore, in every test, please precisely pipette required amount of Diluent and make double dilution in a new container. The prepared working dilution can't be frozen.)
4. Detection Reagent A and Detection Reagent B - Briefly spin or centrifuge the stock Detection A and Detection B before use. Dilute to the working concentration with working Assay Diluent A or B, respectively (1:100).
5. Wash Solution - Dilute 20 mL of Wash Solution concentrate (30x) with 580 mL of deionized or distilled water to prepare 600 mL of Wash Solution (1x).
6. TMB substrate - Aspirate the needed dosage of the solution with sterilized tips and do not dump the residual solution into the vial again.

Note:

1. Making serial dilution in the wells directly is not permitted. o
2. Prepare standard within 15 minutes before assay. Please do not dissolve the reagents at 37 °C directly.
3. Please carefully reconstitute Standards or working Detection Reagent A and B according to the instruction, and avoid foaming and mix gently until the crystals are completely dissolved. To minimize imprecision caused by pipetting, use small volumes and ensure that pipettors are calibrated. It is recommended to suck more than 10μL for once pipetting.
4. The reconstituted Standards, Detection Reagent A and Detection Reagent B can be used only once.
5. If crystals have formed in the Wash Solution concentrate (30x), warm to room temperature and mix gently until the crystals are completely dissolved.
6. Contaminated water or container for reagent preparation will influence the detection result.

Aufbereitung der Proben:

• It is recommended to use fresh samples without long storage, otherwise protein degradation and denaturationmay occur in these samples, leading to false results. Samples should therefore be stored for a short periodat 2 - 8 °C or aliquoted at -20 °C (≤1 month) or -80 °C (≤ 3 months). Repeated freeze-thawcycles should be avoided. Prior to assay, the frozen samples should be slowly thawed and centrifuged toremove precipitates.
• If the sample type is not specified in the instructions, a preliminary test is necessary to determinecompatibility with the kit.
• If a lysis buffer is used to prepare tissue homogenates or cell culture supernatant, there is a possibilityof causing a deviation due to the introduced chemical substance.The recommended dilution factor is for reference only.
• Please estimate the concentration of the samples before performing the test. If the values are not in therange of the standard curve, the optimal sample dilution for the particular experiment has to be determined.Samples should then be diluted with PBS (pH =7.0-7.2).

Testpräzision: Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level of target were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level of target were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: CV < 10%
Inter-Assay: CV < 12%

Beschränkungen: Nur für Forschungszwecke einsetzbar

Handhabung

Vorsichtsmaßnahmen: The Stop Solution suggested for use with this kit is an acid solution. Wear eye, hand, face, and clothing protection when using this material.

Lagerung: 4 °C/-20 °C

Informationen zur Lagerung:

1. For unopened kit: All reagents should be stored according to the labels on the vials. The Standard, Detection Reagent A, Detection Reagent B, and 96-well Strip Plate should be stored at -20 °C upon receipt, while the other reagents should be stored at 4 °C.
2. For opened kits: the remaining reagents must be stored according to the above storage conditions. In addition, please return the unused wells to the foil pouch containing the desiccant and seal the foil pouch with the zipper.

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Haltbarkeit: 6 months

Referenzen für PDPK1 ELISA Kit (ABIN6953358)

Tang, Yang, Wang, Gu, Xia, Feng, Yang, Zhao, Su, Su, Wang: "Nucleoside reverse transcriptase inhibitor-induced rat oocyte dysfunction and low fertility mediated by autophagy." in: Oncotarget, Vol. 9, Issue 3, pp. 3895-3907, (2018) (PubMed).

Details zu PDPK1

Target: PDPK1 (3-phosphoinositide Dependent Protein Kinase-1 (PDPK1))

Andere Bezeichnung: Phosphoinositide Dependent Protein Kinase 1 (PDPK1) (PDPK1 Produkte)

Synonyme: pdpk1 ELISA Kit, MGC82080 ELISA Kit, PDK1 ELISA Kit, PDPK2 ELISA Kit, PRO0461 ELISA Kit, Pdk1 ELISA Kit, zgc:153787 ELISA Kit, 3'-phosphoinositide-dependent protein kinase 1 ELISA Kit, 3-PHOSPHOINOSITIDE-DEPENDENT PROTEIN KINASE 1 ELISA Kit, ATPDK1 ELISA Kit, AtPDK1 ELISA Kit, T32M21.110 ELISA Kit, T32M21_110 ELISA Kit, zgc:77318 ELISA Kit, 3-phosphoinositide dependent protein kinase 1 ELISA Kit, 3-phosphoinositide dependent protein kinase 1 L homeolog ELISA Kit, 3-phosphoinositide dependent protein kinase-1 ELISA Kit, 3-phosphoinositide dependent protein kinase 1a ELISA Kit, 3-phosphoinositide-dependent protein kinase 1 ELISA Kit, 3'-phosphoinositide-dependent protein kinase 1 ELISA Kit, 3-phosphoinositide dependent protein kinase 1b ELISA Kit, PDPK1 ELISA Kit, pdpk1.L ELISA Kit, pdpk1 ELISA Kit, Pdpk1 ELISA Kit, pdpk1a ELISA Kit, LOC100380750 ELISA Kit, pdk-1 ELISA Kit, PDK1 ELISA Kit, pdpk1b ELISA Kit

Pathways: PI3K-Akt Signalweg, T-Zell Rezeptor Signalweg, Fc-epsilon Rezeptor Signalübertragung, EGFR Signaling Pathway, Neurotrophin Signalübertragung, Regulation of Leukocyte Mediated Immunity, Positive Regulation of Immune Effector Process, Cell-Cell Junction Organization, Regulation of Cell Size, Skeletal Muscle Fiber Development, CXCR4-mediated Signaling Events, Signaling Events mediated by VEGFR1 and VEGFR2, VEGFR1 Specific Signals