Description
Human SNAI1 (Snail family transcriptional repressor 1) ELISA Kit
The Human SNAI1 (Snail Family Transcriptional Repressor 1) ELISA Kit is specially designed for the accurate and reliable detection of SNAI1 levels in human samples including serum, plasma, and cell culture supernatants. With its high sensitivity and specificity, this kit ensures precise and reproducible results, making it an excellent tool for various research applications.SNAI1 is a transcriptional repressor that plays a critical role in regulating epithelial-mesenchymal transition (EMT) and promoting tumor metastasis.
Its aberrant expression has been linked to various cancers and is considered a potential target for cancer therapy. This ELISA kit allows researchers to analyze SNAI1 levels in biological samples, leading to a better understanding of its role in disease progression and the development of novel treatment strategies.
| Product Name: | Human SNAI1 (Snail family transcriptional repressor 1) ELISA Kit |
| Product Code: | HUFI06431 |
| Size: | 96 Assays |
| Alias: | rotein sna ELISA Kit, Protein snail homolog 1 ELISA Kit, SLUGH2 ELISA Kit, SNA ELISA Kit, SNAH ELISA Kit, SNAI1 ELISA Kit, snail homolog 1 ELISA Kit, Drosophila ELISA Kit, SNAIL1 ELISA Kit, Zinc finger protein SNAI1 ELISA Kit |
| Detection method: | Sandwich ELISA, Double Antibody |
| Application: | This immunoassay kit allows for the in vitro quantitative determination of Human SNAI1 (Snail family transcriptional repressor 1) concentrations in serum plasma and other biological fluids. |
| Sensitivity: | < 18.75pg/ml |
| Range: | 31.25-2000pg/ml |
| Storage: | 4°C for 6 months |
| Note: | For Research Use Only |
| Recovery: | Matrices listed below were spiked with certain level of Human SNAI1 (Snail family transcriptional repressor 1) and the recovery rates were calculated by comparing the measured value to the expected amount of Human SNAI1 (Snail family transcriptional repressor 1) in samples. | ||||||||||||||||
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| Linearity: | The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Human SNAI1 (Snail family transcriptional repressor 1) and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected. | ||||||||||||||||
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| CV(%): | Intra-Assay: CV<8% Inter-Assay: CV<10% |
| Component | Quantity | Storage |
| ELISA Microplate (Dismountable) | 8×12 strips | 4°C for 6 months |
| Lyophilized Standard | 2 | 4°C/-20°C |
| Sample/Standard Dilution Buffer | 20ml | 4°C |
| Biotin-labeled Antibody(Concentrated) | 120ul | 4°C (Protect from light) |
| Antibody Dilution Buffer | 10ml | 4°C |
| HRP-Streptavidin Conjugate(SABC) | 120ul | 4°C (Protect from light) |
| SABC Dilution Buffer | 10ml | 4°C |
| TMB Substrate | 10ml | 4°C (Protect from light) |
| Stop Solution | 10ml | 4°C |
| Wash Buffer(25X) | 30ml | 4°C |
| Plate Sealer | 5 | - |
Other materials and equipment required:
- Microplate reader with 450 nm wavelength filter
- Multichannel Pipette, Pipette, microcentrifuge tubes and disposable pipette tips
- Incubator
- Deionized or distilled water
- Absorbent paper
- Buffer resevoir
| UniProt Protein Function: | Snail1: a protein of the C2H2-type zinc-finger family that regulates transcription. Involved in embryonic mesoderm formation. Plays a role in the epithelial to mesenchymal transition (EMT). EMT is characterized by decreased levels of the epithelial markers E-cadherin and alpha- and beta-catenin, and increased cellular migration and expression of the mesenchymal markers fibronectin, vimentin, N-cadherin and smooth muscle alpha-actin.E Snail binds to 3 E-boxes of the E-cadherin gene promoter and represses its transcription. NF-kappaB binds and regulates the snail promoter and is critical for EMT, and inhibition of NF-kappaB activity lowered snail levels and morphologically reversed the EMT. |
| UniProt Protein Details: | Protein type:C2H2-type zinc finger protein; Transcription factor Chromosomal Location of Human Ortholog: 20q13.2 Cellular Component: cytoplasm; nucleus Molecular Function:kinase binding; protein binding Biological Process: epithelial to mesenchymal transition; mesoderm formation; negative regulation of DNA damage response, signal transduction by p53 class mediator; negative regulation of transcription from RNA polymerase II promoter; osteoblast differentiation; positive regulation of cell migration; positive regulation of transcription, DNA-dependent |
| NCBI Summary: | The Drosophila embryonic protein snail is a zinc finger transcriptional repressor which downregulates the expression of ectodermal genes within the mesoderm. The nuclear protein encoded by this gene is structurally similar to the Drosophila snail protein, and is also thought to be critical for mesoderm formation in the developing embryo. At least two variants of a similar processed pseudogene have been found on chromosome 2. [provided by RefSeq, Jul 2008] |
| UniProt Code: | O95863 |
| NCBI GenInfo Identifier: | 12644089 |
| NCBI Gene ID: | 6615 |
| NCBI Accession: | O95863.2 |
| UniProt Secondary Accession: | O95863,Q9P113, Q9UBP7, Q9UHH7, B2R842, |
| UniProt Related Accession: | O95863 |
| Molecular Weight: | 29,083 Da |
| NCBI Full Name: | Zinc finger protein SNAI1 |
| NCBI Synonym Full Names: | snail family transcriptional repressor 1 |
| NCBI Official Symbol: | SNAI1 |
| NCBI Official Synonym Symbols: | SNA; SNAH; SNAIL; SLUGH2; SNAIL1; dJ710H13.1 |
| NCBI Protein Information: | zinc finger protein SNAI1 |
| UniProt Protein Name: | Zinc finger protein SNAI1 |
| UniProt Synonym Protein Names: | Protein snail homolog 1; Protein sna |
| Protein Family: | Protein snail |
| UniProt Gene Name: | SNAI1 |
| UniProt Entry Name: | SNAI1_HUMAN |
*Note: Protocols are specific to each batch/lot. For the correct instructions please follow the protocol included in your kit.
Before adding to wells, equilibrate the SABC working solution and TMB substrate for at least 30 min at 37 °C. When diluting samples and reagents, they must be mixed completely and evenly. It is recommended to plot a standard curve for each test.
| Step | Protocol |
| 1. | Set standard, test sample and control (zero) wells on the pre-coated plate respectively, and then, record their positions. It is recommended to measure each standard and sample in duplicate. Wash plate 2 times before adding standard, sample and control (zero) wells! |
| 2. | Aliquot 0.1ml standard solutions into the standard wells. |
| 3. | Add 0.1 ml of Sample / Standard dilution buffer into the control (zero) well. |
| 4. | Add 0.1 ml of properly diluted sample ( Human serum, plasma, tissue homogenates and other biological fluids.) into test sample wells. |
| 5. | Seal the plate with a cover and incubate at 37 °C for 90 min. |
| 6. | Remove the cover and discard the plate content, clap the plate on the absorbent filter papers or other absorbent material. Do NOT let the wells completely dry at any time. Wash plate X2. |
| 7. | Add 0.1 ml of Biotin- detection antibody working solution into the above wells (standard, test sample & zero wells). Add the solution at the bottom of each well without touching the side wall. |
| 8. | Seal the plate with a cover and incubate at 37 °C for 60 min. |
| 9. | Remove the cover, and wash plate 3 times with Wash buffer. Let wash buffer rest in wells for 1 min between each wash. |
| 10. | Add 0.1 ml of SABC working solution into each well, cover the plate and incubate at 37 °C for 30 min. |
| 11. | Remove the cover and wash plate 5 times with Wash buffer, and each time let the wash buffer stay in the wells for 1-2 min. |
| 12. | Add 90 µL of TMB substrate into each well, cover the plate and incubate at 37 °C in dark within 10-20 min. (Note: This incubation time is for reference use only, the optimal time should be determined by end user.) And the shades of blue can be seen in the first 3-4 wells (with most concentrated standard solutions), the other wells show no obvious color. |
| 13. | Add 50 µL of Stop solution into each well and mix thoroughly. The color changes into yellow immediately. |
| 14. | Read the O.D. absorbance at 450 nm in a microplate reader immediately after adding the stop solution. |
When carrying out an ELISA assay it is important to prepare your samples in order to achieve the best possible results. Below we have a list of procedures for the preparation of samples for different sample types.
| Sample Type | Protocol |
| Serum | If using serum separator tubes, allow samples to clot for 30 minutes at room temperature. Centrifuge for 10 minutes at 1,000x g. Collect the serum fraction and assay promptly or aliquot and store the samples at -80°C. Avoid multiple freeze-thaw cycles. If serum separator tubes are not being used, allow samples to clot overnight at 2-8°C. Centrifuge for 10 minutes at 1,000x g. Remove serum and assay promptly or aliquot and store the samples at -80°C. Avoid multiple freeze-thaw cycles. |
| Plasma | Collect plasma using EDTA or heparin as an anticoagulant. Centrifuge samples at 4°C for 15 mins at 1000 × g within 30 mins of collection. Collect the plasma fraction and assay promptly or aliquot and store the samples at -80°C. Avoid multiple freeze-thaw cycles. Note: Over haemolysed samples are not suitable for use with this kit. |
| Urine & Cerebrospinal Fluid | Collect the urine (mid-stream) in a sterile container, centrifuge for 20 mins at 2000-3000 rpm. Remove supernatant and assay immediately. If any precipitation is detected, repeat the centrifugation step. A similar protocol can be used for cerebrospinal fluid. |
| Cell culture supernatant | Collect the cell culture media by pipette, followed by centrifugation at 4°C for 20 mins at 1500 rpm. Collect the clear supernatant and assay immediately. |
| Cell lysates | Solubilize cells in lysis buffer and allow to sit on ice for 30 minutes. Centrifuge tubes at 14,000 x g for 5 minutes to remove insoluble material. Aliquot the supernatant into a new tube and discard the remaining whole cell extract. Quantify total protein concentration using a total protein assay. Assay immediately or aliquot and store at ≤ -20 °C. |
| Tissue homogenates | The preparation of tissue homogenates will vary depending upon tissue type. Rinse tissue with 1X PBS to remove excess blood & homogenize in 20ml of 1X PBS (including protease inhibitors) and store overnight at ≤ -20°C. Two freeze-thaw cycles are required to break the cell membranes. To further disrupt the cell membranes you can sonicate the samples. Centrifuge homogenates for 5 mins at 5000xg. Remove the supernatant and assay immediately or aliquot and store at -20°C or -80°C. |
| Tissue lysates | Rinse tissue with PBS, cut into 1-2 mm pieces, and homogenize with a tissue homogenizer in PBS. Add an equal volume of RIPA buffer containing protease inhibitors and lyse tissues at room temperature for 30 minutes with gentle agitation. Centrifuge to remove debris. Quantify total protein concentration using a total protein assay. Assay immediately or aliquot and store at ≤ -20 °C. |
| Breast Milk | Collect milk samples and centrifuge at 10,000 x g for 60 min at 4°C. Aliquot the supernatant and assay. For long term use, store samples at -80°C. Minimize freeze/thaw cycles. |
