Human/Mouse Phospho-p38 alpha (T180/Y182) ELISA

RayBio® Human/Mouse Phospho-p38 alpha (T180/Y182) ELISA Kit. This assay semi-quantitatively measures phosphorylated p38 alpha (Thr180/Tyr182) in lysate samples.

Specifications

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

• 1. Prepare all reagents and samples as instructed in the manual.

• 2. Add 100 µl of sample or positive control to each well.

• 3. Incubate 2.5 h at RT or O/N at 4 °C.

• 4. Add 100 µl of prepared primary antibody to each well.

• 5. Incubate 1 h at RT.

• 6. Add 100 µl of prepared 1X HRP-Streptavidin to each well.

• 7. Incubate 1 h at RT.

• 8. Add 100 µl of TMB One-Step Substrate Reagent to each well.

• 9. Incubate 30 min at RT.

• 10. Add 50 µl of Stop Solution to each well.

• 11. Read at 450 nm immediately.

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), 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.

Citations

Bizargity P, Liu K, Wang L, Hancock WW, Visner GA. Inhibitory Effects of Pirfenidone on Dendritic Cells and Lung Allograft Rejection. Transplantation. 2012;94(2):114-122. doi:10.1097/TP.0b013e3182584879.

Species: Mouse

Sample Type: Cell Lysate (Lipopolysaccharide and allogeneic treated mice bone marrow DC)

Catalog #: PEL-P38-T180-1 -- 1 Plate Kit

Catalog #: PEL-P38-T180-2 -- 2 Plate Kit

Catalog #: PEL-P38-T180-5 -- 5 Plate Kit

Please email us at cs@medikonia.com for any enquiry. To place an order, please include the catalog number(s) of the product(s) in the email.

Manual

Safety Data Sheet