SMAD5 ELISA Kit

Catalog No. ABIN5526703

Product Details SMAD5 ELISA Kit

Target: SMAD5 (SMAD, Mothers Against DPP Homolog 5 (SMAD5))

Binding Specificity: pSer463, pSer465, total

Reactivity: Human

Detection Method: Colorimetric

Method Type: Cell ELISA

Application: ELISA

Purpose: Cell-Based ELISA Kit. This assay semi-quantitatively measures SMAD5 phosphorylated at Serine-463/465 as well as total SMAD5 in adherent cell lines.

Sample Type: Adherent Cell Culture

Analytical Method: Semi-Quantitative

Specificity: This ELISA kit recognizes Human SMAD5 phosphorylated at site Serine-463/465 as well as total SMAD5.

Characteristics:

• 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

Components:

• 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

Material not included:

• 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

Application Details

Application Notes: Optimal working dilution should be determined by the investigator.

Plate: Pre-coated

Protocol:

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.

Restrictions: For Research Use only

Handling

Storage: -20 °C

Storage Comment: 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.

Expiry Date: 6 months

Target Details for SMAD5

Target: SMAD5 (SMAD, Mothers Against DPP Homolog 5 (SMAD5))

Alternative Name: SMAD5 (SMAD5 Products)

Synonyms: DWFC ELISA Kit, JV5-1 ELISA Kit, MADH5 ELISA Kit, Madh5 ELISA Kit, SBN ELISA Kit, fb67b04 ELISA Kit, mad ELISA Kit, madh5 ELISA Kit, sb:cb400 ELISA Kit, smd5 ELISA Kit, wu:fb67b04 ELISA Kit, SMAD5 ELISA Kit, DKFZp468M0619 ELISA Kit, 1110051M15Rik ELISA Kit, AI451355 ELISA Kit, Dwf-C ELISA Kit, MusMLP ELISA Kit, SMAD family member 5 ELISA Kit, SMAD5 ELISA Kit, Smad5 ELISA Kit, smad5 ELISA Kit

Gene ID: 4090

UniProt: Q99717