SMAD4 ELISA Kit

Catalog No. ABIN4886294

Product Details SMAD4 ELISA Kit

Target: SMAD4 (SMAD Family Member 4 (SMAD4))

Binding Specificity: pThr277, total

Reactivity: Human

Detection Method: Colorimetric

Method Type: Sandwich ELISA

Application: ELISA

Purpose: Human Phospho-SMAD4 (Thr277) and Total SMAD4 ELISA Kit. This assay semi-quantitatively measures phophorylated SMAD4 (Thr277) and Total SMAD4 in lysate samples.

Sample Type: Cell Lysate, Tissue Lysate

Analytical Method: Semi-Quantitative

Specificity: This ELISA kit recognizes Human SMAD4 phosphorylated at site Threonine-277 as well as total SMAD4.

Characteristics:

• Simultaneously measure Phosphorylated protein and pan protein in one experiment (for normalization purpose)
• Screen numerous different cell lysates without performing a Western Blot analysis
• Minimal hands-on time, convenient, and non-radioactive material

Components:

• Pre-Coated 96-well Strip Microplate
• Wash Buffer
• Anti-Phospho Antibody
• Anti-Pan Antibody
• HRP-Conjugated Secondary Antibody
• Streptavidin-Conjugated HRP
• Assay Diluent
• TMB One-Step Substrate
• Stop Solution
• Lysis Buffer
• Positive Control Sample

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

Sample Volume: 100 μL

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 SMAD4

Target: SMAD4 (SMAD Family Member 4 (SMAD4))

Alternative Name: SMAD4 (SMAD4 Products)

Synonyms: smad4 ELISA Kit, DPC4 ELISA Kit, JIP ELISA Kit, MADH4 ELISA Kit, MYHRS ELISA Kit, AW743858 ELISA Kit, D18Wsu70e ELISA Kit, Madh4 ELISA Kit, XSmad4alpha ELISA Kit, Xsmad4 ELISA Kit, dpc4 ELISA Kit, jip ELISA Kit, madh4 ELISA Kit, xsmad4a ELISA Kit, SMAD family member 4 ELISA Kit, Smad4 protein ELISA Kit, SMAD family member 4, gene 1 L homeolog ELISA Kit, SMAD4 ELISA Kit, smad4 ELISA Kit, Smad4 ELISA Kit, smad4.1.L ELISA Kit

Gene ID: 4089

UniProt: Q13485

Pathways: Cell Division Cycle, Chromatin Binding, Autophagy