Nori Human PACAP ELISA Kit
Price range: $508.00 through $916.00
This ELISA kit is for quantification of PACAP in human. This is a quick ELISA assay that reduces time to 50% compared to the conventional method, and the entire assay only takes 3 hours. This assay employs the quantitative sandwich enzyme immunoassay technique and uses biotin-streptavidin chemistry to improve the performance of the assays. An antibody specific for PACAP has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any PACAP present is bound by the immobilized antibody. After washing away any unbound substances, a detection antibody specific for PACAP is added to the wells. Following wash to remove any unbound antibody reagent, a detection reagent is added. After intensive wash a substrate solution is added to the wells and color develops in proportion to the amount of PACAP bound in the initial step. The color development is stopped, and the intensity of the color is measured.
Alternative names for PACAP: Pituitary adenylate cyclase-activating polypeptide, PACAP1, ADCYAP1
This product is for laboratory research use only not for diagnostic and therapeutic purposes or any other purposes.
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Description
Nori Human PACAP ELISA Kit Summary
Alternative names for PACAP: Pituitary adenylate cyclase-activating polypeptide, PACAP1, ADCYAP1
| Assay Type | Solid Phase Sandwich ELISA |
| Format | 96-well Microplate or 96-Well Strip Microplate |
| Method of Detection | Colorimetric |
| Number of Targets Detected | 1 |
| Target Antigen Accession Number | P18509 |
| Assay Length | 3 hours |
| Quantitative/Semiquantitative | Quantitative |
| Sample Type | Plasma, Serum, Cell Culture, Urine, Cell/Tissue Lysates, Synovial Fluid, BAL, |
| Recommended Sample Dilution (Plasma/Serum) | No dilution for sample <ULOQ; sufficient dilution for samples >ULOQ |
| Sensitivity | 6 pg/mL |
| Detection Range | 31.25-2000 pg/mL |
| Specificity | Human PACAP |
| Cross-Reactivity | < 0.5% cross-reactivity observed with available related molecules, < 50% cross-species reactivity observed with species tested. |
| Interference | No significant interference observed with available related molecules |
| Storage/Stability | 4 ºC for up to 6 months |
| Usage | For Laboratory Research Use Only. Not for diagnostic or therapeutic use. |
| Additional Notes | The kit allows for use in multiple experiments. |
Standard Curve
Kit Components
1. Pre-coated 96-well Microplate
2. Biotinylated Detection Antibody
3. Streptavidin-HRP Conjugate
4. Lyophilized Standards
5. TMB One-Step Substrate
6. Stop Solution
7. 20 x PBS
8. Assay Buffer
Other Materials Required but not Provided:
1. Microplate Reader capable of measuring absorption at 450 nm
2. Log-log graph paper or computer and software for ELISA data analysis
3. Precision pipettes (1-1000 µl)
4. Multi-channel pipettes (300 µl)
5. Distilled or deionized water
Protocol Outline
1. Prepare all reagents, samples and standards as instructed in the datasheet.
2. Add 100 µl of Standard or samples to each well and incubate 1 h at RT.
3. Add 100 µl of Working Detection Antibody to each well and incubate 1 h at RT.
4. Add 100 µl of Working Streptavidin-HRP to each well and incubate 20 min at RT.
5. Add 100 µl of Substrate to each well and incubate 5-30 min at RT.
6. Add 50 µl of Stop Solution to each well and read at 450 nm immediately.
Background:
Pituitary adenylate cyclase-activating polypeptide (PACAP) or PACAP1 is a protein that is encoded by the ADCYAP1 gene.[1] PACAP is similar to vasoactive intestinal peptide and stimulate enterochromaffin-like cells. It binds to vasoactive intestinal peptide receptor and to the PACAP receptor. PACAP is mediated by adenylate cyclase-activating polypeptide 1 receptors and stimulates adenylate cyclase and subsequently increases the cAMP level in target cells. PACAP1 not only is a hypophysiotropic hormone, but also functions as a neurotransmitter and neuromodulator. In addition, it plays a role in paracrine and autocrine regulation of certain types of cells. PACP1 gene has been associated with post-traumatic stress disorder (PTSD) in women (but not men).[2] This disorder involves a maladaptive psychological response to traumatic, i.e. existence-threatening, events. Ressler et al. identified an association of a SNP in the gene coding for pituitary adenylate cyclase-activating polypeptide (PACAP), implicating this peptide and its receptor (PAC1) in PTSD. PACP interacts with secretin receptor.[3]
References
- Hosoya M, et al. (1992). “Structure of the human pituitary adenylate cyclase activating polypeptide (PACAP) gene”. Biochim Biophys Acta. 1129 (2): 199–206.
- Ressler, KJ; et al. (2011). “Post-traumatic stress disorder is associated with PACAP and the PAC1 receptor”. Nature. 470 (7335): 492–7.
- Felley, C P; et al. (1992). “Chief cells possess a receptor with high affinity for PACAP and VIP that stimulates pepsinogen release”. Am. J. Physiol. UNITED STATES. 263 (6 Pt 1): G901–7.
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