4E-BP1 [pT46] Phospho-ELISA Kit, Human

Catalog number: KHO0691

Novex™  Related applications: Protein Assays and Analysis

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Description

The 4E-BP1 [pT46] ELISA kit is designed to detect and quantify the level of 4E-BP1 protein phosphorylated at threonine residue 46. This assay is intended for the detection of 4E-BP1 [pT46] from mouse, human and rat cell extracts using 96-well plates and a microplate reader.

Easy to run
More sensitive than Westerns
Convenient precoated, removable 8-well strips

Easy-to-Run Sandwich ELISA
A monoclonal capture antibody specific for 4E-BP1 has been coated onto the wells of the 96-well plate provided. During the first incubation, standards of known 4E-BP1 [pT46] content and unknown samples are pipetted into the wells and the 4E-BP1 antigen binds to the immobilized (capture) antibody. After washing, a rabbit antibody specific for 4E-BP1 phosphorylated at threonine 46 is added to the wells. During the second incubation, this antibody serves as a detection antibody by binding to the immobilized 4E-BP1 protein captured during the first incubation. After washing, a horseradish peroxidase labeled anti-rabbit IgG is added. This binds to the detection antibody to complete the four member sandwich. After a third incubation and washing to remove all the unbound enzyme, a substrate solution (TMB) is added, which is acted upon by the bound enzyme to produce color. The intensity of this colored product is directly proportional to the concentration of 4E-BP1 [pT46] present in the original specimen and the optical density can be read on a standard microplate reader. The total assay incubation time is only 4 hours.

Detect Low Expressing Proteins and Use Less Sample
ELISA kits designed to measure intracellular signaling targets are 2–10X more sensitive than Western blotting. The improved sensitivity enables you to detect low-expressing proteins that otherwise may not be distinguishable from background. In addition, the amount of sample needed to run the assay is less than what is needed for Western blots.

Ready-to-Use 4E-BP1 [pT46] ELISA Kit Contains These Reagents, Buffers, and Supplies:

• 4E-BP1 Antibody-Coated 96-Well Plate
• 4E-BP1 [pT46] Standard
• Standard Diluent Buffer
• 4E-BP1 [pT46] Detection Antibody
• anti-rabbit IgG-HRP (100X)
• HRP Diluent
• Wash Buffer Concentrate (25X)
• Stabilized Chromogen, TMB
• Stop Solution
• Plate Covers
• Detailed protocol with validation tests

Convenient Kit Format
The kit comes with a capture antibody precoated to the plate, eliminating an overnight coating process. The plates are machine coated to provide low variability well-to-well. Furthermore, the plate comes as removable 8-well strips allowing you to run as few samples as you want. In addition, the standards provided in the kit can be used to obtain quantitative results from the standard curve and/or can be used as a positive control.

Rigorous Validation
Each ELISA kit is validated for sensitivity, specificity, precision, and lot-to-lot consistency. See protocol insert for more information on validation.

Part of the ELISA Kit Family
We have a wide array of ELISA kits targeted to disease-related proteins. With excellent sensitivity, proven specificity and consistent performance, these optimized ELISA kits offer you a choice when it comes to getting physiologically relevant and quantitative results.

Learn more about ELISA kits.
Learn more about other immunoassays.
For Research Use Only. Not for use in diagnostic procedures.

Specifications

Accession Number: NM_004095.3, NM_007918.3, NM_053857.1, NP_004086.1, NP_031944.3, NP_446309.1
Gene Aliases: 4e-bp1, AA959816, PHAS-I, PHAS-I, 4E-BP1, 4EBP1, BP-1, MGC4316, PHAS-I
Shipping Condition: Wet Ice
Concentration: 1.6 U⁄ml
Detection Method: Colorimetric
For Use With (Equipment): Microplate Reader
Gene ID: 116636, 13685, 1978
Gene Symbol: EIF4EBP1
Incubation Time: 4 hrs
Label or Dye: HRP (Horseradish Peroxidase)
Marker: 4E-BP1
Product Size: 96 assays
Protein Family: Transcription Factors
Research Category: Signal Transduction
Sample Type (Specific): Cell Extracts
Sample Volume: 10 µl
Sensitivity: <0.8 U⁄mL
Species: Human, Mouse, Rat
Target Gene: eukaryotic translation initiation factor 4E binding protein 1

Contents & storage

Pre-coated plate(s), standard or calibrator, detector antibody, HRP conjugate, diluents, wash buffer, substrate(s), and stop solution. Store kit at 2-8°C. See product manual for detailed contents and storage conditions for maximum stability.

Documents

Manuals & protocols