eIF4EBP1 (EIF4EBP1)
(Eukaryotic Translation Initiation Factor 4E Binding Protein 1 (EIF4EBP1))
Bindungsspezifität
pThr36, total
Reaktivität
Maus, Human
Nachweismethode
Colorimetric
Methodentyp
Cell ELISA
Applikation
ELISA
Verwendungszweck
Human and Mouse Phospho-4E-BP1 (Thr36) and Total 4E-BP1 Cell-Based ELISA Cell-Based ELISA Kit. This assay semi-quantitatively measures 4E-BP1 phosphorylated at Threonine-36 as well as total 4E-BP1 in adherent cell lines.
Proben
Adherent Cell Culture
Analytische Methode
Semi-Quantitative
Spezifität
This ELISA kit recognizes Human and Mouse 4E-BP1 phosphorylated at site Threonine-36 as well as total 4E-BP1.
Produktmerkmale
Rapidly measure phosphorylated protein in adherent cell lines
Simultaneously measure Phosphorylated protein and pan protein in one experiment (for normalization purpose)
No sample lysis is needed
Compatible with a standard ELISA plate reader
Bestandteile
Uncoated 96-well Strip Microplate
Wash Buffers
Fixing Solution
Quenching Buffer
Blocking Buffer
Anti-phospho antibody
Anti-pan antibody
HRP-Conjugated Secondary Antibody
TMB One-Step Substrate
Stop Solution
Benötigtes Material
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
Optimal working dilution should be determined by the investigator.
Plattentyp
Pre-coated
Protokoll
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.
Beschränkungen
Nur für Forschungszwecke einsetzbar
Lagerung
-20 °C
Informationen zur Lagerung
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), HRP-Streptavidin (Item G), 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. Reconstituted Positive Control (Item K) should be stored at -70 °C.