photo-ELISA Kit
photo-ELISA Kit

Nori Human Sirtuin 4 ELISA Kit

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DataSheet   

This ELISA kit is for quantification of sirtuin 4 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 sirtuin 4 has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any sirtuin 4 present is bound by the immobilized antibody. After washing away any unbound substances, a detection antibody specific for sirtuin 4 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 sirtuin 4 bound in the initial step. The color development is stopped, and the intensity of the color is measured.

Alternative names for Sirtuin 4: SIRT4, Sirtuin 4, SIR2L4

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 Sirtuin 4 ELISA Kit Summary

Alternative names for Sirtuin 4: SIRT4, Sirtuin 4, SIR2L4

 

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 Q9Y6E7
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 Human Sirtuin 4
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: 

Sirtuin 4 (SIRT4) is a mitochondrial protein which is encoded by the SIRT4 gene.[1] SIRT4 is member of the mammalian sirtuin family of proteins, which are homologs to the yeast Sir2 protein. SIRT4 exhibits NAD+-dependent deacetylase activity. SIRT4 is a mitochondrial ADP-ribosyltransferase that inhibits mitochondrial glutamate dehydrogenase 1 activity, thereby downregulating insulin secretion in response to amino acids.[2] A deacetylation of malonyl-CoA decarboxylase enzyme by SIRT4 represses the enzyme activity, inhibiting fatty acid oxidation in muscle and liver cells.[3][4] SIRT4 has a suppressive effect on peroxisome proliferator-activated receptor alpha (PPAR-α) which downregulates fatty acid oxidation in liver cells.[9] Deacetylation of ADP/ATP translocase 2 (ANT2) increases cellular ATP by dampening mitochondrial uncoupling.[4] SIRT4 is a mitochondrial tumor suppressor protein.[9] Overexpression of SIRT4 inhibits cancer cell proliferation by inhibition of glutamine metabolism.[4][5]

References

  1. Frye RA (1999). Biochem. Biophys. Res. Commun. 260 (1): 273–79.
  2. Haigis MC, et al. (2006). Cell. 126 (5): 941–54. doi:1016/j.cell.2006.06.057.
  3. Nasrin N, et al. (2010). J. Biol. Chem. 285 (42): 31995–32002. doi:1074/jbc.M110.124164.
  4. Li S, Zheng W (2018). Progress in Molecular Biology and Translational Science. 154: 147–168.
  5. Yoo HC, et al. (2020). Experimental & Molecular Medicine. 52 (9): 1496–1516.

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