RayBiotech/人/小鼠磷酸-4E-BP1（T36）ELISA/1平板试剂盒/PEL-4EBP1-T36-1
商品编号：
PEL-4EBP1-T36-1
品牌：
RayBiotech
市场价：
¥0.00
美元价：
0.00
产品分类：
抗小鼠
公司分类：
Anti_mouse
联系Q Q：
3392242852
电话号码：
4000-520-616
电子邮箱：
info@ebiomall.com
商品介绍
Antigen Information
Accession Number:
- Q13541
Gene ID:
- 1978
Gene Symbols:
- EIF4EBP1
Protein Name & Synonyms:
Eukaryotic translation initiation factor 4E-binding protein 1 (4E-BP1) (eIF4E-binding protein 1) (Phosphorylated heat- and acid-stable protein regulated by insulin 1) (PHAS-I)
Assay Format
Pathway:
- AMPK Signaling
- PI3K-AKT Signaling
- Translation
Specificity:
The antibody pair provided in this kit recognizes human and mouse 4E-BP1 phosphorylated at site Threonine-36.
Number of Targets Detected:
1
Species Detected:
- Human
- Mouse
Compatible Sample Types:
- Cell Lysates
- Tissue Lysates
Design Principle:
- Sandwich-based
Method of Detection:
Colorimetric
Quantitative/Semi-Quantitative:
- Semi-Quantitative
Solid Support:
96-well Microplate
Product Specifications
Size:
1, 2, or 5 x 96-Well Strip Microplate Kit
Introduction
RayBio® Phospho-4E-BP1 (Thr36) ELISA kit is a very rapid, convenient and sensitive assay kit that can monitor the activation or function of important biological pathways in human and mouse cell lysates. By determining 4E-BP1 protein in your experimental model system, you can verify pathway activation in your cell lysates. You can simultaneously measure numerous different cell lysates without spending excess time and effort in performing a Western Blotting analysis.This Sandwich ELISA kit is an in vitro enzyme-linked immunosorbent assay for the measurement of human and mouse phospho-4E-BP1. An anti-pan 4E-BP1 antibody has been coated onto a 96-well plate. Samples are pipetted into the wells and 4E-BP1 present in a sample is bound to the wells by the immobilized antibody. The wells are washed and rabbit anti-4E-BP1 (Thr36) antibody is used to detect phosphorylated 4E-BP1. After washing away unbound antibody, HRP-conjugated anti-rabbit IgG 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 4E-BP1 (Thr36) bound. The Stop Solution changes the color from blue to yellow, and the intensity of the color is measured at 450 nm.
Product Features
- Rapidly measure phosphorylated protein in lysates
- Screen numerous different cell lysates without performing a Western Blot analysis
- Minimal hands-on time, convenient, and non-radioactive material
Application Notes
Kit Components
- Pre-Coated 96-well Strip Microplate
- Wash Buffer
- Anti-Phospho Antibody
- HRP-Conjugated Secondary Antibody
- Assay Diluent
- TMB One-Step Substrate
- Stop Solution
- Lysis Buffer
- Positive Control Sample
Other Materials Required
- Distilled or deionized water
- 100 ml and 1 liter graduated cylinders
- Tubes to prepare sample dilutions
- Protease and Phosphatase inhibitors
- Precision pipettes to deliver 2 µl to 1 ml volumes
- Adjustable 1-25 ml pipettes for reagent preparation
- Benchtop rocker or shaker
- Microplate reader capable of measuring absorbance at 450 nm
Protocol Outline
- Prepare all reagents and samples as instructed in the manual.
- Add 100 µl of sample or positive control to each well.
- Incubate 2.5 h at RT or O/N at 4 °C.
- Add 100 µl of prepared primary antibody to each well.
- Incubate 1 h at RT.
- Add 100 µl of prepared 1X HRP-Streptavidin to each well.
- Incubate 1 h at RT.
- Add 100 µl of TMB One-Step Substrate Reagent to each well.
- Incubate 30 min at RT.
- Add 50 µl of Stop Solution to each well.
- Read at 450 nm immediately.
Typical Data
Positive Control
HeLa cells were treated with FBS. Solubilize cells at 4 x 107 cells/ml in Cell Lysate Buffer. Serial dilutions of lysates were analyzed in this ELISA. Please see step 3 of Part VI Reagent Preparation for detail. 
Insulin Stimulation of 293 Cell Line
293 cells were treated or untreated with Insulin. Cell lysates were analyzed using this phosphoELISA and Western Blot. 
Storage/Stability
Upon receipt, the kit should be stored at –20°C. Please use within 6 months from the date of shipment. After initial use, Wash Buffer Concentrate (Item B), Assay Diluent (Item E), TMB One-Step Substrate Reagent (Item H), Stop Solution (Item I) and Cell Lysate Buffer (Item J) should be stored at 4°C to avoid repeated freeze-thaw cycles. Return unused wells to the pouch containing desiccant pack, reseal along entire edge, and store at –20°C. Item D, store at 2-8°C for up to one month (store at -20°C for up to 6 months, avoid repeated freeze-thaw cycles). Reconstituted Positive Control (Item K) should be stored at -70°C.
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