The hippocampus undergoing different treatments were homogenized using PBS and centrifuged at 8000 g at 4°C for 15 min to harvest the supernatant. Afterwards, the levels of M1 pro-inflammatory cytokines interleukin (IL)-1β (ab255730) and tumor necrosis factor-α (TNF-α) (ab236712), and M2 anti-inflammatory cytokines IL-10 (ab214566) and IL-4 (ab100771, all from Abcam) were analyzed using the specific ELISA kit and calculated according to the standard curve [21 (link)].
Ab214566
Manufactured by Abcam
Sourced in United States
Ab214566 is a monoclonal antibody that targets the human EGFR protein. It is designed for use in various immunoassay applications.
Automatically generated - may contain errors
Lab products found in correlation
Ab236712, by Abcam (7 mentions)
Ab255730, by Abcam (4 mentions)
Softmax pro software, by Molecular Devices (4 mentions)
Versamax, by Molecular Devices (3 mentions)
Ab234570, by Abcam (2 mentions)
Ab269547, by Abcam (2 mentions)
Elx808, by Agilent Technologies (1 mentions)
Ab46070, by Abcam (1 mentions)
Ab46107, by Abcam (1 mentions)
Ab219045, by Abcam (1 mentions)
Ab239425, by Abcam (1 mentions)
Ab119558, by Abcam (1 mentions)
Mak066, by Merck Group (1 mentions)
S0024, by Beyotime (1 mentions)
Mak085, by Merck Group (1 mentions)
Ab234579, by Abcam (1 mentions)
Ab263883, by Abcam (1 mentions)
Milliplex map rat cytokine chemokine magnetic bead panel kit, by Merck Group (1 mentions)
Sekr 0071, by Solarbio (1 mentions)
Rat tnf α elisa kit, by Abcam (1 mentions)
12 protocols using ab214566
1
Hippocampal Cytokine Profiling
2
Serum Cytokine Detection by ELISA
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3
Inflammatory Marker ELISA Quantification
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Inflammation markers including interferon gamma (IFN-γ; ab46107), monocyte chemoattractant protein-1 (MCP-1; ab219045), tumor necrosis factor alpha (TNF-α; ab239425), and interleukin 10 (IL-10; ab214566) ELISA kits were obtained from Abcam Inc. (Cambridge, UK) and executed as stated in the manufacturers’ instructions.
4
Cytokine Levels in Plasma Analysis
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The levels of the cytokines IL-6 (R6000B, R&D Systems, MN, USA), IL-10 (ab214566, Abcam, MA, USA), and TNF-α (ab236712, Abcam, Cambridge, MA, USA) in plasma were measured using ELISA kits according to the manufacturer’s instructions. The optical density at 450 nm was measured using a microplate reader (VersaMax with SoftMax Pro software, Molecular Devices, San Jose, CA, USA).
5
Quantitative Cytokine and Inflammasome Profiling
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Kits purchased from Abcam, Waltham, USA (CAT # ab119558 and ab214566), were utilized for assessment of tissue TGF-β1 and IL10 levels, respectively. IFN-α levels were quantified using kits supplied by CUSABIO, Houston, TX 77054, USA (CAT #CSB-E08637r). Assay of NLRP3 levels was executed using kits provided by Aviva Systems Biology Co., San Diego, CA 92121, USA (CAT # OKCD04232). Determination of the fore-mentioned parameters was carried out following the vendor’s protocol.
6
Cytokine Profiling in Plasma
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The levels of the cytokines interleukin (IL)-6 (R6000B, R&D Systems, Inc., Minneapolis, MN, USA), IL-10 (ab214566, Abcam, MA, USA), and TNF-α (ab236712, Abcam, MA, USA) in plasma were measured using ELISA kits, in accordance with the manufacturer’s instructions. The optical density at 450 nm was measured using a VersaMax microplate reader (SoftMax Pro software, Molecular Devices, CA, USA).
7
Cytokine Levels in Spleen and Plasma
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The levels of the cytokines IL-6 (R6000B, R&D Systems, MN, USA), IL-10 (ab214566, Abcam, MA, USA), and TNF-α (ab236712, Abcam, MA, USA) in spleen homogenates and plasma were measured using ELISA kits according to the manufacturer’s instructions. To assess cytokine levels in the spleen, frozen tissue was homogenized in RIPA buffer with protease inhibitor cocktail. The optical density at 450 nm was detected by a microplate reader (VersaMax with SoftMax Pro software, Molecular Devices, CA, USA).
8
Liver Function and Inflammatory Markers
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Enzyme-linked immunosorbent assay (ELISA) kits of AST (ab263883, abcam, USA), ALT (ab234579, abcam, USA), tumor necrosis factor (TNF)-α (ab236712, abcam, USA), interleukin (IL)-1β (ab255730, abcam, USA), IL-6 (ab234570, abcam, USA) and IL-10 (ab214566, abcam, USA), and LDH kit (MAK066, Merck, German) were used to detect the levels of liver function markers and inflammatory factors in the serum according to the manufacturer’s instructions.
After washing the liver with a normal saline solution, 100 mg of liver tissue was cut and made into a 10% homogenate. The homogenate was centrifuged at 2,500 r/minute for 15 minutes to obtain the supernatant. The malondialdehyde (MDA) kit (MAK085, Merck, German), superoxide dismutase (SOD) kit (MM-0386R2, MEIMIAN, China), and nitric oxide (NO) kit (S0024, Beyotime, China) were used to detect the levels of MDA, SOD, and NO, respectively, in the supernatant.
After washing the liver with a normal saline solution, 100 mg of liver tissue was cut and made into a 10% homogenate. The homogenate was centrifuged at 2,500 r/minute for 15 minutes to obtain the supernatant. The malondialdehyde (MDA) kit (MAK085, Merck, German), superoxide dismutase (SOD) kit (MM-0386R2, MEIMIAN, China), and nitric oxide (NO) kit (S0024, Beyotime, China) were used to detect the levels of MDA, SOD, and NO, respectively, in the supernatant.
9
Serum cytokine analysis of UC-MSCs therapy
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The serum cytokines of rats were analyzed at 1, 3, and 7 days after hUC-MSCs injection. About 200 μl blood was collected through intraocular canthus from each rat, and then centrifuged at 7500 RPM (Thermo Scientific Legend Microro 17R, Rotor.75003424) for 15 min to get serum. The serum was prepared with coagulant tubes centrifugation. All the serum was stored in −80°C and was performed with immunoassay to measure cytokine levels later. The serum multiplex immunoassays were performed to measure the pro-inflammatory factors (tumor necrosis factor alpha [TNF-α], interferon gamma [IFN-γ], interleukin [IL]-1β, IL-6) in the serum of each group at 1, 3, and 7 days after UC-MSCs injection, using MILLIPLEX MAP Rat Cytokine/Chemokine Magnetic Bead Panel kit # RECYTMAG-65K-04 (Millipore, Saint Louis, MO, USA), based on the xMAP Luminex technology. ELISA assays were performed to measure the anti-inflammatory factors (IL-10 and IL-13) using the rat IL-10 (ab214566, Abcam, Inc) and IL-13 ELISA kit (ab269547, Abcam, Inc), according to the manufacturer’s instructions.
10
Cytokine Profiling in Respiratory Conditions
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IL-8 (SEKR-0071; Solarbio, Beijing, China), IL-6 (ab234570; Abcam, MA, USA), IL-4 (ab100770; Abcam, MA, USA), IL-10 (ab214566; Abcam, MA, USA), and TNF-α (ab236712; Abcam, MA, USA) in peripheral venous blood and BALF were measured using ELISA kits in accordance with the manufacturer's instructions. The levels of cytokines IL-1β (ab255730; Abcam, MA, USA) in the supernatants of rPMVECs were also tested.
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