Human MAGED1 (MAGE family member D1) ELISA Kit (HUFI03530)
- SKU:
- HUFI03530
- Product Type:
- ELISA Kit
- Size:
- 96 Assays
- Uniprot:
- Q9Y5V3
- Sensitivity:
- 0.094ng/ml
- Range:
- 0.156-10ng/ml
- ELISA Type:
- Sandwich
- Synonyms:
- DLXIN 1, MAGE D1 antigen, MAGE tumor antigen CCF, MAGED1, melanoma antigen family D, 1, Melanoma associated antigen D1, NRAGE, PP2250
- Reactivity:
- Human
Frequently bought together:
Description
Human MAGED1 (MAGE family member D1) ELISA Kit (HUFI03530)
The Human MAGED1 (MAGE Family Member D1) ELISA Kit is specifically designed for the accurate quantification of MAGED1 levels in human serum, plasma, and cell culture supernatants. This kit offers exceptional sensitivity and specificity, ensuring precise and reliable results for a variety of research applications.MAGED1 is a member of the MAGE family of proteins, which have been implicated in various cellular processes including cell growth, differentiation, and apoptosis. Dysregulation of MAGE proteins has been linked to cancer, developmental disorders, and neurological diseases, highlighting the importance of studying MAGED1 levels in research and clinical settings.
With its advanced technology and quality assurance, the Human MAGED1 ELISA Kit is a valuable tool for researchers seeking to investigate the role of MAGED1 in disease pathogenesis and therapy development. Trust in this kit to provide accurate and reproducible measurements for your studies.
| Product Name: | Human MAGED1 (MAGE family member D1) ELISA Kit |
| Product Code: | HUFI03530 |
| Size: | 96 Assays |
| Alias: | DLXIN 1 ELISA Kit, MAGE D1 antigen ELISA Kit, MAGE tumor antigen CCF ELISA Kit, MAGED1 ELISA Kit, melanoma antigen family D ELISA Kit, 1 ELISA Kit, Melanoma associated antigen D1 ELISA Kit, NRAGE ELISA Kit, PP2250 ELISA Kit |
| Detection method: | Sandwich ELISA, Double Antibody |
| Application: | This immunoassay kit allows for the in vitro quantitative determination of Human MAGED1 (MAGE family member D1) concentrations in serum plasma and other biological fluids. |
| Sensitivity: | < 0.094ng/ml |
| Range: | 0.156-10ng/ml |
| Storage: | 4°C for 6 months |
| Note: | For Research Use Only |
| Recovery: | Matrices listed below were spiked with certain level of Human MAGED1 (MAGE family member D1) and the recovery rates were calculated by comparing the measured value to the expected amount of Human MAGED1 (MAGE family member D1) in samples. Enquire for more information. |
| Linearity: | The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Human MAGED1 (MAGE family member D1) and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected. Enquire for more information. |
| 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: | NRAGE: a member of the melanoma antigen gene (MAGE) family. Most of the genes of this family encode tumor specific antigens that are not expressed in normal adult tissues except testis. Although NRAGE shares strong homology with members of the MAGE family, it is expressed in almost all normal adult tissues. Binds p75 neurotrophin receptor (p75NTR) and antagonizes its association with TrkA, inhibits cell cycle progression, and facilitates p75NTR-mediated apoptosis. May act as a regulator of the function of DLX family members. |
| UniProt Protein Details: | Protein type:Adaptor/scaffold Chromosomal Location of Human Ortholog: Xp11.23 Cellular Component: protein complex; cytoplasm; plasma membrane; chromatin; nucleus Molecular Function:protein binding; transcription coactivator activity Biological Process: regulation of apoptosis; regulation of transcription from RNA polymerase II promoter; positive regulation of MAP kinase activity; regulation of transcription, DNA-dependent; nerve growth factor receptor signaling pathway; positive regulation of apoptosis; positive regulation of transcription, DNA-dependent; circadian regulation of gene expression; regulation of circadian rhythm; negative regulation of transcription, DNA-dependent; negative regulation of epithelial cell proliferation |
| NCBI Summary: | This gene is a member of the melanoma antigen gene (MAGE) family. Most of the genes of this family encode tumor specific antigens that are not expressed in normal adult tissues except testis. Although the protein encoded by this gene shares strong homology with members of the MAGE family, it is expressed in almost all normal adult tissues. This gene has been demonstrated to be involved in the p75 neurotrophin receptor mediated programmed cell death pathway. Three transcript variants encoding two different isoforms have been found for this gene. [provided by RefSeq, Jul 2008] |
| UniProt Code: | Q9Y5V3 |
| NCBI GenInfo Identifier: | 62906893 |
| NCBI Gene ID: | 9500 |
| NCBI Accession: | Q9Y5V3.3 |
| UniProt Related Accession: | Q9Y5V3 |
| Molecular Weight: | |
| NCBI Full Name: | Melanoma-associated antigen D1 |
| NCBI Synonym Full Names: | MAGE family member D1 |
| NCBI Official Symbol: | MAGED1 |
| NCBI Official Synonym Symbols: | NRAGE; DLXIN-1 |
| NCBI Protein Information: | melanoma-associated antigen D1 |
| UniProt Protein Name: | Melanoma-associated antigen D1 |
| UniProt Synonym Protein Names: | MAGE tumor antigen CCF; MAGE-D1 antigen; Neurotrophin receptor-interacting MAGE homolog |
| UniProt Gene Name: | MAGED1 |
| UniProt Entry Name: | MAGD1_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. |
Related Products
Human Kinesin Family, Member 5A (KIF5A) ELISA Kit (HUDL01610)
€649
Human Kinesin Family, Member 1A (KIF1A) ELISA Kit (HUDL01608)
€649
Human SLFNL11 (Schlafen family member 11) ELISA Kit (AEFI00126)
€599
Human Kinesin Family, Member 20A (KIF20A) ELISA Kit (HUDL01609)
€649
Human Golgin A8 Family, Member A (GOLGA8A) ELISA Kit (HUDL01222)
€749
Human Kinesin Family, Member 18A (KIF18A) ELISA Kit (HUDL01607)
€649
Human Kinesin Family, Member 14 (KIF14) ELISA Kit (HUDL01606)
€649
Human Kinesin Family, Member 5B (KIF5B) ELISA Kit (HUDL01611)
€649