IGF1 levels in normal culture medium collected after 48 hr from different cells were detected with the Human IGF1 ELISA Kit (ab100545; Abcam, CA, USA) according to the manufacturer's instructions.
Human igf1 elisa kit
Manufactured by Abcam
Sourced in United States, Spain
The Human IGF1 ELISA Kit is a quantitative sandwich enzyme immunoassay designed for the measurement of IGF1 (Insulin-like Growth Factor 1) in human serum, plasma, and cell culture supernatants. The kit utilizes a human IGF1-specific antibody coated on a microplate and a biotin-conjugated anti-human IGF1 antibody for detection.
Automatically generated - may contain errors
Lab products found in correlation
Dimension autoanalyzer, by Siemens (2 mentions)
α tubulin, by Merck Group (1 mentions)
T9026, by Merck Group (1 mentions)
Nitrocellulose membrane, by Bio-Rad (1 mentions)
Radioimmunoassay, by Thermo Fisher Scientific (1 mentions)
Vacutainer, by BD (1 mentions)
Human cxcl5 ena 78 quantikine elisa kit, by R&D Systems (1 mentions)
Opti mem 1 1x reduced serum medium, by Thermo Fisher Scientific (1 mentions)
Human gdnf elisa kit, by Abcam (1 mentions)
Human egf elisa kit, by Abcam (1 mentions)
Fluostar omega, by BMG Labtech (1 mentions)
Hbm msc, by Lonza (1 mentions)
9 protocols using human igf1 elisa kit
1
Quantifying IGF1 Levels in Cell Culture
2
Fasting Serum Biomarkers Assessment
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Fasting serum samples were obtained after centrifugation for 15 min at 1800× g. Serum total cholesterol, high-density lipoprotein cholesterol (HDL-c), triglycerides, and glucose concentrations were determined by means of a Dimension Autoanalyzer (Dade Behring Inc., Deerfield, IL, USA). Low-density lipoprotein cholesterol (LDL-c) was calculated using the Friedewald equation [16 (link)]. Insulin levels were measured by a radioimmunoassay method using BioSource International Inc. (Camarillo, CA, USA). HOMA-IR was estimated using the following equation: HOMA-IR = fasting insulin (μIU/mL) × fasting glucose (mM)/22.5 [17 (link)]. Serum vitamin B12 was quantified by chemiluminescent immunoassay (Roche Diagnostics GmbH, Penzberg, Germany). C-reactive protein (CRP) was measured in a Dimension Autoanalyzer (Dade Behring Inc., Deerfield, IL, USA). Insulin growth factor type 1 (IGF-1) was determined using a Human IGF1 ELISA Kit (Abcam, Madrid, Spain). Carcinoembryonic antigen (CEA) and carbohydrate antigen 19.9 (CA 19.9) were measured by ELISA (DRG diagnostics, Marburg, Germany).
3
Western Blot and ELISA for IGF1 Pathway
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells were lysed in RIPA buffer containing a mixture of protease inhibitors. Protein lysates were resolved by SDS/PAGE and transferred to nitrocellulose membranes (Bio-Rad). Membranes were blocked using 5% BSA for 1 h at room temperature, followed by primary antibody incubation overnight at 4 °C. The primary antibodies used were the following: IGF1R (Abcam, ab16817 and Novus, 100-91823), MGMT (Abcam, ab39253), and α-tubulin (Sigma, T9026). After washing with 1x TBST buffer, membranes were incubated with species-specific secondary antibodies for 1 h at room temperature. Protein band intensity was quantified using ImageJ software.
IGF1 concentration in conditioned media was assessed using Human IGF1 ELISA kit (Abcam) based on manufacturer's instruction.
IGF1 concentration in conditioned media was assessed using Human IGF1 ELISA kit (Abcam) based on manufacturer's instruction.
4
Measuring Plasma IGF-1 Levels with ELISA
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The Human IGF-1 ELISA Kit (ab100545; Abcam, Cambridge, MA, USA) was used to measure plasma levels of IGF-1 which were converted to ng/mL.
5
Comprehensive Metabolic and Hormonal Profile
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Fasting blood samples were obtained from the antecubital vein and placed in vacutainer tubes (BD Vacutainer™, Franklin Lakes, NJ, USA). Serum glucose, total cholesterol, triglycerides, and HDL cholesterol (HDL-c) were measured in a Dimension auto analyzer (Dade Behring Inc., Deerfield, IL, USA) through enzymatic methods (Randox Laboratories Ltd., Crumlin, UK). The LDL cholesterol (LDL-c) was calculated using the Friedewald equation [25 (link)]. Insulin was quantified by radioimmunoassay (BioSource International, Camarillo, CA, USA). Corrected calcium was assessed using a complex metric method from Boehringer Hitachi 717. Alkaline phosphatase was calculated using ELISA (LifeSpan Biosciences Inc., Madrid, Spain). Insulin growth factor type 1 (IGF-1) was determined using Human IGF1 ELISA Kit (Abcam, Madrid, Spain). The homeostasis model assessment of insulin resistance (HOMA-IR) was calculated using the following equation: HOMA-IR = fasting insulin (IU/mL) x fasting glucose (mmol/L)/22.5 [26 (link)].
6
Quantification of IGF-1 in HepG2 cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
HepG2 cells treated for 24 h with CM collected from PHT cells either control (non-coding siRNA), RAPTOR-silenced (inhibition of mTORC1), and RICTOR-silenced (inhibition of mTOR C2) PHT cells were used for the quantitative measurement of IGF-1 using human IGF-1 ELISA kit (Abcam). IGF-1 was also measured in CM from PHT cells with either control, RAPTOR-, or RICTOR-silenced PHT cells before treatment of HepG2 cells.
7
IGF-1 Protein Quantification by ELISA
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Treated cell proteins were isolated by washing them with PBS and adding 50 µL of Complete M-Lysis according to schedule. The cell was shaken for 10 minutes. The lysed cell was transferred to a new tube and kept at −80° C . Briefly, 100 µL of each standard solution and sample was added to the well to then shaken and incubated at 4°C overnight. After overnight incubation, the samples were consecutively incubated with a 1x detection antibody, HRP-Streptavidin, and TMB one-step substrate with washing between each step. The measurement of IGF-1 concentration was conducted at a wavelength of 450 nm. Measurement of IGF-1 protein was performed using the methods listed on the human IGF-1 ELISA kit (Abcam).
8
Quantification of Cytokines in Cell Supernatant
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Quantitation of IL30, CXCL5 and IGF1, in the supernatant derived from murine or human PC cells, was carried out using the following ELISA kits, according to manufacturer’s protocols: Human CXCL5/ENA-78 Quantikine ELISA Kit (#DX000, R&D Systems, Minneapolis, MN, USA); human IGF1 ELISA Kit (#ab211651, Abcam, Cambridge, UK); and mouse Interleukin-27 subunit alpha ELISA Kit (#CSB-E08466m, Cusabio, Wuhan, China).
9
Secretome Analysis of Labeled hBM-MSCs
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
hBM-MSCs (Lonza) were cultured at 75-cm2 flasks for five passages and labeled with Molday ION Rhodamine B™ and CellTracker™ Green CMFDA or transfected with mRNA GFP. To test the release of EGF, GDNF, IGF-1, and HGF, the concentration (or amount) of these proteins was measured in media taken from native and labeled cells cultured in serum-free Opti-Mem® I (1x) Reduced Serum Medium (Life Technologies) for 24 h. At the 2nd, 5th, and 7th day after hBM-MSC staining, the medium was collected and thickened by centrifugation (30 min, 3000×g, RT) in Vivaspin® concentrators with PES 5000 or 10,000 MWCO membranes (Sartorius). Then, the protein content was analyzed using the Bradford method with absorbance level read on micro plate reader FLUOstar Omega (BMG Labtech) at 450 nm. The assessment of human EGF, GDNF, IGF-1, and HGF secretion into serum-free medium was performed using specific ELISA kits: human EGF ELISA Kit, human GDNF ELISA Kit; human IGF1 ELISA Kit, and human HGF ELISA Kit (Abcam) following the manufacturer’s instructions with 100 μg of total protein placed on each well of 96-well ELISA plate.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!