PIK3CA ELISA Kit

Catalog No. ABIN6974973

Product Details PIK3CA ELISA Kit

Target: PIK3CA (Phosphoinositide-3-Kinase, Catalytic, alpha Polypeptide (PIK3CA))

Reactivity: Mouse

Detection Method: Colorimetric

Method Type: Sandwich ELISA

Detection Range: 23.5 pg/mL - 1500 pg/mL

Minimum Detection Limit: 23.5 pg/mL

Application: ELISA

Purpose: For the quantitative determination of mouse phosphotylinosital 3 kinase (PI3K) concentrations in serum, plasma, tissue homogenates.

Sample Type: Plasma, Serum, Tissue Homogenate

Analytical Method: Quantitative

Specificity: This assay has high sensitivity and excellent specificity for detection of mouse PI3K. No significant cross-reactivity or interference between mouse PI3K and analogues was observed. Note: Limited by current skills and knowledge, it is impossible for us to complete the cross-reactivity detection between mouse PI3K and all the analogues, therefore, cross reaction may still exist.

Sensitivity: 5.8 pg/mL

Components:

• Assay plate
• Standard
• HRP-avidin (100 x concentrate)
• Biotin-antibody (100 x concentrate)
• Sample Diluent
• HRP-avidin Diluent
• Biotin-antibody Diluent
• Wash Buffer (25 x concentrate)
• TMB Substrate
• Stop Solution
• Adhesive Strip

Application Details

Application Notes: Optimal working dilution should be determined by the investigator.

Sample Volume: 100 μL

Assay Time: 1 - 4.5 h

Plate: Pre-coated

Protocol:

1. Prepare reagents, samples and standards as instructed.
2. Add 100 µL standard or sample to each well. Incubate 2 hours at 37 °C.
3. Remove the liquid of each well, don't wash.
4. Add 100 µL Biotin-antibody (1x) to each well. Incubate 1 hour at 37 °C.
5. Aspirate and wash 3 times.
6. Add 100 µL HRP-avidin (1x) to each well. Incubate 1 hour at 37 °C
7. Aspirate and wash 5 times.
8. Add 90 μL of TMB Substrate to each well. Incubate for 15-30 minutes at 37 °C. Protect from light.
9. Add 50 µL Stop Solution to each well. Read at 450 nm within 5 minutes.

Reagent Preparation:

1. Biotin-antibody (1x) - Centrifuge the vial before opening. Biotin-antibody requires a 100-fold dilution. A suggested 100-fold dilution is 10 μL of Biotin-antibody + 990 μL of Biotin-antibody Diluent.
2. HRP-avidin (1x) - Centrifuge the vial before opening. HRP-avidin requires a 100-fold dilution. A suggested 100-fold dilution is 10 μL of HRP-avidin + 990 μL of HRP-avidin Diluent.
3. Wash Buffer(1x)- If crystals have formed in the concentrate, warm up to room temperature and mix gently until the crystals have completely dissolved. Dilute 20 mL of Wash Buffer Concentrate (25 x) into deionized or distilled water to prepare 500 mL of Wash Buffer (1 x).
4. Standard Centrifuge the standard vial at 6000-10000rpm for 30s. Reconstitute the Standard with 1.0 mL of Sample Diluent. Do not substitute other diluents. This reconstitution produces a stock solution of 1500 pg/mL. Mix the standard to ensure complete reconstitution and allow the standard to sit for a minimum of 15 minutes with gentle agitation prior to making dilutions. Pipette 250 μL of Sample Diluent into each tube (S0-S6). Use the stock solution to produce a 2-fold dilution series (below). Mix each tube thoroughly before the next transfer. The undiluted Standard serves as the high standard (1500 pg/mL). Sample Diluent serves as the zero standard (0 pg/mL).

Note:

• Kindly use graduated containers to prepare the reagent. Please don't prepare the reagent directly in the Diluent vials provided in the kit.
• Bring all reagents to room temperature (18-25 °C) before use for 30 min.
• Prepare fresh standard for each assay. Use within 4 hours and discard after use.
• Making serial dilution in the wells directly is not permitted.
• Please carefully reconstitute Standards according to the instruction, and avoid foaming and mix gently until the crystals have completely dissolved. To minimize imprecision caused by pipetting, use small volumes and ensure that pipettors are calibrated. It is recommended to suck more than 10 μL for once pipetting.
• Distilled water is recommended to be used to make the preparation for reagents. Contaminated water or container for reagent preparation will influence the detection result.

Sample Preparation:

• 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).

Assay Precision: Intra-assay Precision (Precision within an assay): CV%<8% Three samples of known concentration were tested twenty times on one plate to assess.
Inter-assay Precision (Precision between assays): CV%<10% Three samples of known concentration were tested in twenty assays to assess.

Restrictions: For Research Use only

Handling

Storage: 4 °C,-20 °C

Storage Comment: Unopened kit Store at 2 - 8°C. Do not use the kit beyond the expiration date. May be stored for up to 1 month at 2 - 8°C. Coated assay Try to keep it in a sealed aluminum foil bag, plate and avoid the damp. Standard May be stored for up to 1 month at 2 - 8° C. If Biotin-antibody don't make recent use, better keep it store at HRP-avidin -20°C. Biotin-antibody Diluent Opened kit HRP-avidin Diluent Sample May be stored for up to 1 month at 2 - 8°C. Diluent Wash Buffer TMB Substrate Stop Solution *Provided this is within the expiration date of the kit.

Expiry Date: 6 months

References for PIK3CA ELISA Kit (ABIN6974973)

Song, Ju, Xiao, Wang, Liu, Fu, Liu, Gu, Li, Cheng: "Application of multifunctional targeting epirubicin liposomes in the treatment of non-small-cell lung cancer." in: International journal of nanomedicine, Vol. 12, pp. 7433-7451, (2018) (PubMed).

Shin, Choi, Kim, Yu, Shin, Choi, Roh: "High doses of caffeine reduce in vivo osteogenic activity in prepubertal rats." in: Journal of anatomy, Vol. 227, Issue 1, pp. 10-20, (2015) (PubMed).

Target Details for PIK3CA

Target: PIK3CA (Phosphoinositide-3-Kinase, Catalytic, alpha Polypeptide (PIK3CA))

Alternative Name: phosphoinositide-3-kinase, catalytic, alpha polypeptide (PIK3CA Products)

Synonyms: CLOVE ELISA Kit, CWS5 ELISA Kit, MCAP ELISA Kit, MCM ELISA Kit, MCMTC ELISA Kit, PI3K ELISA Kit, p110-alpha ELISA Kit, C-P3K ELISA Kit, PI3-kinase ELISA Kit, PI3K-alpha ELISA Kit, pi3-k ELISA Kit, pi3k ELISA Kit, AA414921 ELISA Kit, C530050K14 ELISA Kit, p50alpha ELISA Kit, p55alpha ELISA Kit, p85alpha ELISA Kit, 6330412C24Rik ELISA Kit, caPI3K ELISA Kit, p110 ELISA Kit, p110alpha ELISA Kit, phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic subunit alpha ELISA Kit, Phosphoinositide-3-kinase, catalytic, alpha polypeptide ELISA Kit, phosphatidylinositol-4,5-bisphosphate 3-kinase, catalytic subunit alpha ELISA Kit, phosphoinositide-3-kinase regulatory subunit 1 ELISA Kit, PIK3CA ELISA Kit, GL50803_14855 ELISA Kit, pik3ca ELISA Kit, Pik3ca ELISA Kit, Pik3r1 ELISA Kit

Background:

Abbreviation: PIK3CA

Alias: MGC142161, MGC142163, PI3K, p110-alpha, PI3-kinase p110 subunit alpha|PtdIns-3-kinase p110|phosphatidylinositol 3-kinase, catalytic, 110-KD, alpha|phosphatidylinositol 3-kinase, catalytic, alpha pol

UniProt: P42337

Pathways: PI3K-Akt Signaling, RTK Signaling, TCR Signaling, AMPK Signaling, Interferon-gamma Pathway, TLR Signaling, Fc-epsilon Receptor Signaling Pathway, EGFR Signaling Pathway, Neurotrophin Signaling Pathway, Inositol Metabolic Process, Hepatitis C, CXCR4-mediated Signaling Events, Signaling Events mediated by VEGFR1 and VEGFR2, Signaling of Hepatocyte Growth Factor Receptor, VEGFR1 Specific Signals, VEGF Signaling