Nori Rat CTLA4 ELISA Kit
Price range: $508.00 through $916.00
This ELISA kit is for quantification of CTLA4 in rat. 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 CTLA4 has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any CTLA4 present is bound by the immobilized antibody. After washing away any unbound substances, a detection antibody specific for CTLA4 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 CTLA4 bound in the initial step. The color development is stopped, and the intensity of the color is measured.
Alternative names for CTLA4: Cytotoxic T-lymphocyte-associated protein 4 (CTLA4), CD152
This product is for laboratory research use only not for diagnostic and therapeutic purposes or any other purposes.
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Description
Nori Rat CTLA4 ELISA Kit Summary
Alternative names for CTLA4: Cytotoxic T-lymphocyte-associated protein 4 (CTLA4), CD152
| 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 | A0A096MJE4 |
| 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 | 30 pg/mL |
| Detection Range | 0.156-10 ng/mL |
| Specificity | Rat CTLA4 |
| 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:
Cytotoxic T-lymphocyte-associated protein 4 (CTLA4), also known as CD152, is a protein receptor that functions as an immune checkpoint and downregulates immune responses and is encoded by the CTLA4 gene.[1]. CTLA4 is a member of the immunoglobulin superfamily that is expressed by activated T cells and transmits an inhibitory signal to T cells. CTLA4 is homologous to the T-cell co-stimulatory protein, CD28, and both molecules bind to CD80 and CD86, also called B7-1 and B7-2 respectively, on antigen-presenting cells. CTLA4 is constitutively expressed in regulatory T cells but only upregulated in conventional T cells after activation – a phenomenon which is particularly notable in cancers.[2] It acts as an “off” switch when bound to CD80 or CD86 on the surface of antigen-presenting cells. CTLA-4 binds CD80 and CD86 with greater affinity and avidity than CD28 thus enabling it to outcompete CD28 for its ligands. CTLA4 transmits an inhibitory signal to T cells,[3][4] whereas CD28 transmits a stimulatory signal.[5] T cell activation through the T cell receptor and CD28 leads to increased expression of CTLA-4. CTLA-4 may recruit a phosphatase to the T cell receptor (TCR), thus attenuating the signal.[6] More recent work has suggested that CTLA-4 may function in vivo by capturing and removing B7-1 and B7-2 from the membranes of antigen-presenting cells, thus making these unavailable for triggering of CD28.[7] CTLA-4 may also function via modulation of cell motility and/or signaling through PI3 kinase[8] CTLA-4 reverses the TCR-induced ‘stop signal’ needed for firm contact between T cells and antigen-presenting cells (APCs).[9]
References
- Dariavach P, et al. (1988). Eur. J. Immunol. 18 (12): 1901–5.
- Syn, Nicholas L; et al. (2017). The Lancet Oncology. 18 (12): e731–e741.
- Waterhouse P, et al. (1995). Science. 270 (5238): 985–8.
- Krummel MF, Allison JP (1995). J. Exp. Med. 182 (2): 459–65.
- ^ Harding FA, et al. (1992). Nature. 356 (6370): 607–9.
- Lee KM, et al. (1998). Science. 282 (5397): 2263–6.
- Qureshi OS, et al. (2011). Science. 332 (6029): 600–3.
- Knieke K, et al. (2012). PLoS ONE. 7 (3): e31391.
- Rudd CE, et al. (2009). Immunol. Rev. 229 (1): 12–26.
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