Human SMAD4 ELISA Kit
Cat no : KE00360
Synonyms
SMAD4,DPC4,hSMAD4,JIP,MAD homolog 4
Validation Data Gallery
Product Information
KE00360 is a solid phase sandwich Enzyme Linked-Immuno-Sorbent Assay (Sandwich ELISA). The human SMAD4 ELISA kit is to be used to detect and quantify protein levels of endogenous human SMAD4. The assay recognizes human SMAD4. An antibody specific for human SMAD4 has been pre-coated onto the microwells. The human SMAD4 protein in samples is captured by the coated antibody after incubation. Following extensive washing, another horseradish peroxidase (HRP)-conjugated antibody specific for human SMAD4 is added to detect the captured human SMAD4 protein. For signal development, followed by Tetramethyl-benzidine (TMB) reagent. Solution containing sulfuric acid is used to stop color development and the color intensity which is proportional to the quantity of bound protein is measurable at 450 nm with the correction wavelength set at 630 nm.
| Product name | Human SMAD4 ELISA Kit |
| Tests | 1 X 96 well plate |
| Sample type | Serum, Plasma, Cell lysate |
| Assay type | Sandwich |
| Sensitivity | 0.4 pg/mL |
| Range | 39-2500 pg/mL |
| Reactivity | Human |
| Tested applications | Sandwich ELISA |
| Gene ID (NCBI) | 4089 |
Recovery
| Sample Type | Average | Range |
|---|---|---|
| Human serum | 83% | 70%-116% |
| Cell lysate | 111% | 99%-126% |
IntraAssay
| Sample | n | mean ( pg/mL) | SD | CV% |
|---|---|---|---|---|
| 1 | 20 | 1,231.1 | 90.0 | 7.3 |
| 2 | 20 | 222.3 | 23.0 | 10.3 |
| 3 | 20 | 53.3 | 3.1 | 5.8 |
InterAssay
| Sample | n | mean ( pg/mL) | SD | CV% |
|---|---|---|---|---|
| 1 | 24 | 1,236.8 | 82.1 | 6.6 |
| 2 | 24 | 240.3 | 11.4 | 4.7 |
| 3 | 24 | 56.1 | 3.5 | 6.2 |
Background Information
SMAD4, also called SMAD family member 4, Mothers against decapentaplegic homolog 4, or DPC4 (Deleted in Pancreatic Cancer-4) is a highly conserved protein present in all metazoans. It belongs to the SMAD family of transcription factor proteins, which act as mediators of TGF-β signal transduction. The TGF-β family of cytokines regulates critical processes during the lifecycle of metazoans, with important roles during embryo development, tissue homeostasis, regeneration, and immune regulation. SMAD4 interacts with R-Smads, such as SMAD2, SMAD3, SMAD1, SMAD5 and SMAD8 (also called SMAD9) to form heterotrimeric complexes. Transcriptional coregulators, such as WWTR1 (TAZ) interact with SMADs to promote their function. Once in the nucleus, the complex of SMAD4 and two R-SMADS binds to DNA and regulates the expression of different genes depending on the cellular context. Smad4 GSK3 phosphorylations have been proposed to regulate the protein stability during pancreatic and colon cancer progression.
Properties
| Storage Instructions | All the reagents are stored at 2-8℃ for 6 months or -20℃ for 12 months. Refer to the protocol for further storage instructions. |
| Synonyms | SMAD4,DPC4,hSMAD4,JIP,MAD homolog 4 |
FAQ
1、I have limited sample diluent. How should I handle samples requiring high dilution factors?
Answer: We recommend performing a serial dilution. For example, if human serum requires a 1,000,000-fold dilution, you can use a three-step 100× dilution method as follows:
Step 1: Take 2 uL of human serum and add it to 198 uL of sample diluent, mix thoroughly to obtain a 100-fold diluted sample.
Step 2: Take 2 uL of the 100-fold diluted sample and add it to 198 uL of sample diluent, mix thoroughly to obtain a 10,000-fold diluted sample.
Step 3: Take 2 uL of the 10,000-fold diluted sample and add it to 198 uL of sample diluent, mix thoroughly to obtain a 1,000,000-fold diluted sample.
If further dilution is required, additional serial steps can be used. We do not recommend using any other solutions or water for dilution, except for the sample diluent we suggest. If your sample diluent is still insufficient, please contact our customer service for more sample diluent.