Human sirt1 elisa kit

Manufactured by Abcam

Sourced in United Kingdom

The Human SIRT1 ELISA Kit is a quantitative assay designed for the measurement of SIRT1 levels in human cell and tissue samples. It utilizes the sandwich ELISA technique to detect and quantify the target protein.

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Lab products found in correlation

Taqman genotyping assay, by Thermo Fisher Scientific (3 mentions) Multiskan fc microplate photometer, by Thermo Fisher Scientific (3 mentions) Hetasep, by STEMCELL (1 mentions)

6 protocols using human sirt1 elisa kit

Sirt1 Protein Expression in COPD Patients

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Whole blood was collected from 48 patients (Table 3) into 6 ml round-bottom tubes containing EDTA, nucleated cells were separated from red blood cells using Hetasep™ (Stemcell Technologies) according to the manufacture’s protocol, then the pellets of blood cells were stored at −80 °C until measurement of Sirt1 protein expression. The Sirt1 protein expression of the stored blood cells was measured using a human Sirt1 ELISA kit (Abcam). The Sirt1 protein expressions in blood cells were referenced to the protein concentration measured by the Bradford method.Table 3

Characteristics of Patients who measured Sirt1 protein expression

Characteristics	Smoking control	Mild COPD	Moderate COPD	Severe COPD
GOLD	-	Stage I	Stage II	Stage III, IV
Number of patients	12	5	20	11
Age (years)	69.6 ± 6.4	74.2 ± 5.5	75.9 ± 7.7	74.2 ± 7.6
Gender (Male/Female)	(12/0)	(4/1)	(19/1)	(10/1)
FVC predict (%)	99.2 ± 15.6	114.1 ± 5.4	97.2 ± 11.6	81.9 ± 17.5*
FEV1/FVC (%)	77.1 ± 7.6	59.4 ± 7.6*	51.0 ± 9.4*	32.7 ± 6.7*
FEV1% predict	96.6 ± 14.3	88.2 ± 8.0	63.5 ± 7.3*	34.2 ± 9.2*
Smoking (Pack years)	48.1 ± 31.7	43.0 ± 11.8	56.3 ± 31.7	68.3 ± 32.9
Current smoker (no.)	3	0	6	0
Medication (no.)
LABA	-	4	17	10
LAMA	-	3	16	8
ICS	-	2	6	7

Values are expressed as means ± SD. *p < 0.05 vs smoking control

Kato R., Mizuno S., Kadowaki M., Shiozaki K., Akai M., Nakagawa K., Oikawa T., Iguchi M., Osanai K., Ishizaki T., Voelkel N.F, & Toga H. (2016). Sirt1 expression is associated with CD31 expression in blood cells from patients with chronic obstructive pulmonary disease. Respiratory Research, 17, 139.

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Quantification of Astrocyte-Secreted SIRT1

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The level of astrocyte-secreted SIRT1 in the brain compartment (BC) was assessed using the Human SIRT1 ELISA Kit (Abcam, Cambridge, UK, cat. no. ab171573), following the manufacturer’s instructions. Along with biological samples (supernatants from astrocyte cultures), each ELISA microplate contained positive controls (serial dilutions of the SIRT1 standard, serving to construct the standard curve) and negative controls (sample dilution buffer and/or astrocyte culture medium containing or not LPS). Based on standard curves generated for each assay independently, the SIRT1 concentration in the analyzed samples was calculated from absorbance units (Au), measured by the microplate spectrophotometer (ASYS UVM 340 Microplate Reader) at a wavelength of 450 nm. Samples were assayed in duplicate, and the values were averaged. ELISA experiments were performed in triplicate. To obtain a better estimate of data variability in each glycemic background analyzed, the SIRT1 concentration values for time point 0 h were pooled across each experiment, and for time point 12 h, they were pooled for LPS-untreated and LPS-treated groups.

Grabowska A.D., Wątroba M., Witkowska J., Mikulska A., Sepúlveda N, & Szukiewicz D. (2023). Interplay between Systemic Glycemia and Neuroprotective Activity of Resveratrol in Modulating Astrocyte SIRT1 Response to Neuroinflammation. International Journal of Molecular Sciences, 24(14), 11640.

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Genetic and Protein Analysis of SIRT1 in Periodontitis

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DNA samples were extracted from peripheral venous blood using the DNA salting-out method. Genotyping of all three SNPs was performed using TaqMan^® genotyping assays (Applied Biosystems, Foster City, CA, USA) and SIRT1 (rs3818292, rs3758391, and rs7895833) according to the manufacturer’s instructions using real-time polymerase chain reaction (PCR). Serum SIRT1 levels were determined in 500 control subjects and 201 patients with periodontitis. Serum SIRT1 levels in patients were determined using the commercial enzyme-linked immunosorbent assay (ELISA) kit for human SIRT1 (Human SIRT1 ELISA Kit, Abcam, Cambridge, United Kingdom) according to the manufacturer’s instructions, and optical density was measured immediately at a wavelength of 450 nm, using a microplate reader (Multiskan FC microplate photometer, Thermo Scientific, Waltham, MA, USA). The SIRT1 level was calculated using the standard curve; sensitivity range of the standard curve was 0.63–40 ng/mL, sensitivity 132 pg/mL [21 (link)].

Kriaučiūnas A., Liutkevičienė R., Gedvilaitė G., Kaikarytė K., Vilkevičiūtė A., Gleiznys D., Pacauskienė I, & Žekonis G. (2022). Value of Serum Sirtuin-1 (SIRT1) Levels and SIRT1 Gene Variants in Periodontitis Patients. Medicina, 58(5), 653.

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Genetic and Serum Analysis of SIRT1

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Blood samples were collected in vacutainers with EDTA. EDTA acts as an anticoagulant, binding the calcium ions and interrupting the clotting of the blood sample.
After whole blood collection, we allowed the blood to clot by leaving it undisturbed at room temperature for 30 min. The clot was removed by centrifuging at 3,000 × g for 10 min in a refrigerated centrifuge. The resulting supernatant—blood serum was stored in a fridge at -20 degrees C temperature.
We extracted DNA samples from peripheral venous blood using the DNA salting-out method. Genotyping of all three SNPs was performed using TaqMan® genotyping assays (Applied Biosystems Foster City, CA, USA): SIRT1 (rs3818292, rs3758391, and rs7895833) according to the manufacturer's instructions using real-time polymerase chain reaction (PCR). Serum SIRT1 levels were determined in control subjects and patients using the commercial enzyme-linked immunosorbent assay (ELISA) kit for human SIRT1 (Human SIRT1 ELISA Kit, Abcam, Cambridge, United Kingdom) according to the manufacturer's instructions, and optical density was measured immediately at a wavelength of 450 nm using a microplate reader (Multiskan FC microplate photometer, Thermo Scientific, Waltham, MA). The SIRT1 level was calculated using the standard curve; the sensitivity range of the standard curve: 0.63–40 ng/ml, sensitivity 132 pg/ml.

Kubiliute A., Gedvilaite G., Vilkeviciute A., Kriauciuniene L., Bruzaite A., Zaliuniene D, & Liutkeviciene R. (2023). The role of SIRT1 level and SIRT1 gene polymorphisms in optic neuritis patients with multiple sclerosis. Orphanet Journal of Rare Diseases, 18, 64.

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SIRT1 Genotyping and Serum Level Assessment

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We extracted DNA samples from peripheral venous blood using the DNA salting-out method. The genotyping of all three SNPs was performed using TaqMan

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Genotyping assays (Applied Biosystems Foster City, CA, USA): SIRT1 (rs3818292, rs3758391, and rs7895833) according to manufacturer’s instructions using the real-time polymerase chain reaction (PCR) method. SIRT1 serum levels were assessed in 15 control subjects and 24 LSCC patients. The SIRT1 level in serum of LSCC patients was determined using the commercial enzyme-linked immunoassay (ELISA) kit for human SIRT1 (Human SIRT1 ELISA Kit, Abcam, Cambridge, United Kingdom), according to the manufacturer’s instructions, and the optical density was immediately measured at 450 nm wavelength using a microplate reader (Multiskan FC microplate photometer, Thermo Scientific, Waltham, MA)). The SIRT1 level was calculated according to the standard curve; standard curve sensibility range: 0.63–40 ng/mL, sensitivity 132 pg/mL.

Vaiciulis P., Liutkeviciene R., Liutkevicius V., Vilkeviciute A., Gedvilaite G, & Uloza V. (2022). Association of SIRT1 single gene nucleotide polymorphisms and serum SIRT1 levels with laryngeal squamous cell carcinoma patient survival rate. Cancer Biomarkers, 34(2), 175-188.

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Measuring Sirt-1 Expression in 18Co Cells

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Sirt-1 expression was measured in 18Co cells, seeded in 12-well plates and treated as described above, by using Human SIRT1 ELISA kit (Abcam, Cambridge U.K.). Briefly, cell extracts were obtained in cells previously solubilized in Cell Extraction Buffer and centrifugated at 18,000 x g for 20 min at 4 °C according to the manufacturer's instructions. Data have been normalized on total protein content and Sirt-1 expression was expressed as percent of the respective levels measured in control.

Domazetovic V., Bonanomi A.G., Stio M., Vincenzini M.T, & Iantomasi T. (2019). Resveratrol decreases TNFα-induced ICAM-1 expression and release by Sirt-1-independent mechanism in intestinal myofibroblasts. Experimental cell research, 382(2).

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