Protein phosphorylation research
Explore protein phosphorylation through gel-based separation and membrane-based detection. Find the Phosbind reagent and control for your workflow.
01 / PRODUCT SELECTION
Choose acrylamide for gel separation, a biotin reagent for membrane detection, and α-casein to check the gel system.
| Cat. No. | Product | Application | Details |
|---|---|---|---|
| F4002 | Phos binding reagent (Phosbind) acrylamide GEL SEPARATION | Separate phosphorylated and non-phosphorylated proteins by mobility shift in a metal-containing gel. | View product ↗ |
| F4001 | Phos binding reagent (Phosbind) Biotin MEMBRANE DETECTION | Detect phosphorylated proteins on PVDF using a biotin reagent, streptavidin–HRP and chemiluminescence. | View product ↗ |
| F4004 | Phos binding reagent (Phosbind) Biotin LC MEMBRANE DETECTION · LONG LINKER | A long-linker biotin reagent for detecting phosphorylated proteins on PVDF membranes. | View product ↗ |
| F4003 | Dephosphorylated α-Casein from Bovine Milk POSITIVE CONTROL | Positive control for validating the Phosbind SDS-PAGE separation system. Partially dephosphorylated α-casein retains phosphorylated forms. | View product ↗ |
Follow each product’s protocol for preparation and use. Available sizes, prices and batch documents are listed on the product pages.
02 / HOW IT WORKS
Phosphate binding supports two distinct readouts: a shift in gel migration or a signal on a membrane.
F4002 + F4003 POSITIVE CONTROL
Incorporate Phosbind acrylamide into the resolving gel with the appropriate metal ions. Phosphorylated species migrate more slowly, enabling separation from non-phosphorylated species. Use F4003 as a positive control to verify the separation system.
F4001 / F4004
After protein transfer to PVDF, use Phosbind Biotin or Biotin LC for phosphate recognition. Detect the biotin label with streptavidin–HRP and a chemiluminescent substrate.
Use the current F4002 protocol for metal-removal and transfer conditions.
03 / APPLICATION DATA
Gel separation and membrane detection examples from the APExBIO protein biology overview.
Phosphatase-treated protein X resolved by Phosbind SDS-PAGE. The original figure includes assay conditions.
View reagent and protocol ↗Importin α phosphorylation detected with Phosbind Biotin and streptavidin–HRP, with and without Purvalanol A treatment.
View detection reagent ↗04 / RELATED PRODUCT
F4005 · Prestained Protein Marker
(Triple color, EDTA free, 10–250 kDa)
A companion marker for protein electrophoresis. The original comparison shows the marker in a Phosbind gel and a conventional SDS-PAGE gel.
View F4005 ↗05 / FREQUENTLY ASKED QUESTIONS
Selected guidance from the F4002 product FAQ, with F4003 as the positive control.
At 50 μM in a 5 mL resolving gel, 10 mg prepares approximately 66 mini gels; 5 mg prepares 33. Yield changes with gel volume and reagent concentration.
Product FAQ ↗Store F4002 at 4°C, protected from light, for up to one year; prepared stock remains stable for at least three months under these conditions. Prepare Mn²⁺–Phosbind gels fresh.
Product FAQ ↗Choose an EDTA-free marker such as F4005. Phosbind changes protein mobility, so marker positions cannot determine the target’s molecular weight accurately.
Product FAQ ↗After electrophoresis, remove metal ions with EDTA-containing buffer, then rinse with EDTA-free transfer buffer before transfer. See the F4002 protocol for timing and concentrations.
Product FAQ ↗Check sample amount, phosphorylation preservation, metal removal before transfer, transfer efficiency, and antibody/blocking conditions for the gel-based Western workflow.
Product FAQ ↗Use F4003 as a positive control, compare phosphatase-treated samples, and run conventional SDS-PAGE in parallel to check for degradation.
Product FAQ ↗06 / RESOURCES
Consult the detailed protocols for reagent preparation, gel conditions and downstream analysis.
