All reagents, restriction enzymes and protein extraction kit were products of ThermoScientific USA unless otherwise stated. Antibodies against TIMP-2 (R&D Systems; MAB971) and MT1-MMP (Abcam; Ab72685 against activated MT1-MMP from cleavage adjacent to Tyr112) were purchased separately from R&D Systems, MN and Abcam, MA, USA. Pwo DNA polymerase and ProteoExtract® Native Membrane protein extraction kit were products of Merck, Darmstadt, Germany. TIMP-1 and -2 proteins were produced in house using Sf-9 insect cell as described in our previous paper [44 (link)]. AAV293, HeLa and HT1080 cell lines were purchased from the Shanghai Cell Bank, Chinese Academy of Science while Lenti-X 293T cells were imported directly from Takara® BIO Inc., CA without further authentication. Non-obese diabetic/severe combined immunodeficiency (NOD/SCID) mice were ordered through GenePharma Co. Ltd, Biobay, Suzhou Industrial Park, Jiangsu, China. All the experiments in this study have been performed independently at least twice to confirm the reproducibility of the findings.
Mab971
Manufactured by Abcam
Sourced in United States
MAB971 is a monoclonal antibody that recognizes human Fibronectin. It is suitable for use in various immunoassay applications.
Automatically generated - may contain errors
Lab products found in correlation
Protein extraction kit, by Thermo Fisher Scientific (1 mentions)
Pwo dna polymerase, by Merck Group (1 mentions)
Lenti x 293t cells, by Takara Bio (1 mentions)
Proteoextract native membrane protein extraction kit, by Merck Group (1 mentions)
Ab32249, by Abcam (1 mentions)
Ab27794, by Abcam (1 mentions)
Ab16051, by R&D Systems (1 mentions)
Ba1034, by Thermo Fisher Scientific (1 mentions)
Nl004, by R&D Systems (1 mentions)
Mab5718, by R&D Systems (1 mentions)
Ab38898, by Abcam (1 mentions)
Bca kit, by Yeasen (1 mentions)
2 protocols using mab971
1
TIMP-2 and MT1-MMP Protein Extraction
2
Protein Expression Analysis in Trophoblast Cells
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The total proteins of B6Tert, HTR8/SVneo, and JEG-3 cells were collected and cracked by high RIPA buffer (Solarbio, Beijing, China). The concentration of proteins was detected using a BCA kit (Yeasen, Shanghai, China). Then, the SDS-PAGE was used to separate the proteins, and the proteins were transferred to a nitrocellulose (NC) membrane (Haoran, Shanghai, China). The NC membrane was blocked by 5% non-fat milk at room temperature for 1.5 h, and then incubated with the primary antibodies (anti-BRIT1, Abcam, ab121277, dilution: 1: 900; anti-matrix metalloproteinase-2 (MMP-2), R&D, IC903G, dilution: 1: 700; anti-MMP-9, Abcam, ab38898, dilution: 1: 1000; anti-tissue inhibitor of metalloproteinases-1 (TIMP-1), R&D, IC970G, dilution: 1: 700; anti-TIMP-2, R&D, MAB971, dilution: 1: 500; anti-Wnt2, Abcam, ab27794, dilution: 1: 600; anti-Wnt3, Abcam, ab32249, dilution: 1: 600; anti-β-catenin, Abcam, ab16051, dilution: 1: 600; anti-GAPDH, R&D, MAB5718, dilution: 1: 800) at 4°C overnight. Subsequently, the NC membrane was incubated with the secondary antibodies at room temperature for 1.5 h (goat anti-mouse IgG, Abcam, ab6785, 1: 8000; donkey anti-rabbit IgG, R&D, NL004, 1: 5000; mouse anti-rabbit IgG, Invitrogen, BA1034, 1: 7000). Chemiluminescence detection was carried out using ECL reagent (Huiying, Shanghai, China).
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