U-Blot® Acetylated NF-KB p65 and Total NF-KB p65 ELISA Kit
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$999.00
Product details
The Acetylated-NF-KB p65 and Total-NF-KB p65 ELISA Kit is an in vitro enzyme-linked immunosorbent assay for the measurement of human acetylated ATP-citrate synthase and total ATP-citrate synthase. An anti-pan NF-KB p65 antibody has been coated onto a 96-well plate. Samples are pipetted into the wells and NF-KB p65 present in a sample is bound to the wells by the immobilized antibody. The wells are washed and biotinylated anti-acetylation antibody is used to detect acetylated NF-KB p65 or biotinylated anti-NF-KB p65 antibody is used to detect total NF-KB p65 . After washing away unbound antibody, HRP-conjugated streptavidin is pipetted to the wells. The wells are again washed, a TMB substrate solution is added to the wells and color develops in proportion to the amount of acetylated NF-KB p65 or total NF-KB p65 bound. The Stop Solution changes the color from blue to yellow, and the intensity of the color is measured at 450 nm.This assay enables quick and easy quantification of target proteins and allows high-throughput screening of numerous different lysates without the need for traditional Western Blot analysis. It features minimal hands-on operation time, simple workflow, and completely non-radioactive detection system, ensuring safe and convenient experimental operation.
Specifications
| Size | 1 Plate Kit, 2 Plate Kit, 5 Plate Kit |
| Species | Human |
| Compatible Sample Types | Tissue Lysates, Cell Lysates |
| Accession Number | Q04206 |
| Gene Id | 5970 |
| Gene Symbols | RELA, NFKB3 |
| Protein Name / Synonyms | Transcription factor p65; Nuclear factor NF-kappa-B p65 subunit; Nuclear factor of kappa light polypeptide gene enhancer in B-cells 3 |
| Solid Support | 96-well Microplate |
| Design Principle | Sandwich-based |
| Method Of Detection | Colorimetric |
| Research Area | Post-Translational Modifications, NFkB Signaling, Acetylation, Epigenetics |
| Modification | Acetylation |
| Shipping Type | 2‑8 ℃ |
| Storage | -20°C |
| Kit Components | Microplate Coated With Anti-Pan-NF-KB p65 Antibody:1 plate;Anti-Acetylation Detection Antibody:1 vial;Anti-NF-KB p65 Detection Antibody:1 vial;HRP-Conjugated Streptavidin:200 µL;Wash Buffer:25 mL;Assay Diluent:15 mL;Lysis Buffer:5 mL;TMB One-Step Substrate Reagent:12 mL;Stop Solution:8 mL; |
| Additional Materials Required | Microplate reader capable of measuring absorbance at 450 nm;Protease and Phosphatase inhibitors;Precision pipettes to deliver 2 µL to 1 mL volumes;Adjustable 1-25 mL pipettes for reagent preparation;100 mL and 1 liter graduated cylinders;Log-log graph paper or computer and software for ELISA data analysis;Absorbent paper;Tubes to prepare sample dilutions;Distilled or deionized water;Shaker |
| Assay Procedure Summary | Prepare all reagents and samples as instructed; Add 100 µL sample to each well. Incubate 2.5 hours at room temperature or overnight at 4°C with gentle shaking; Add 100 µL prepared detection antibody to each well. Incubate for 1 hour at room temperature with gentle shaking; Add 100 µL prepared HRP-Conjugated Streptavidin solution. Incubate for 1 hour at room temperature with gentle shaking; Add 100 µL TMB One-Step Substrate Reagent to each well. Incubate 30 minutes at room temperature; Add 50 µL Stop Solution to each well. Read at 450 nm immediately. |
| Storage/Stability | The entire kit may be stored at -20°C for up to 6 months from the date of shipment. Avoid repeated freeze-thaw cycles. |
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