Anti-HGF antibody [EPR12230]

• IP

Unknown

Immunoprecipitation - Anti-HGF antibody [EPR12230] (AB178395)

ab178395 (purified) at 1 : 30 dilution (2μg) immunoprecipitating HGF in Human placenta lysate.
Lane 1 (input) : Human placenta lysate 10μg
Lane 2 (+) : ab178395 & Human placenta lysate
Lane 3 (-) : Rabbit monoclonal IgG (ab172730) instead of ab178395 in Human placenta lysate
For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366) was used for detection at 1 : 1000 dilution.
Blocking and diluting buffer : 5% NFDM/TBST.

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Immunoprecipitation - Anti-HGF antibody [EPR12230] (ab178395)

Predicted band size: 83 kDa

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• WB

Lab

Western blot - Anti-HGF antibody [EPR12230] (AB178395)

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Western blot - Anti-HGF antibody [EPR12230] (ab178395) at 1/200 dilution

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Human placenta lysates at 15 µg

Secondary

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Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Predicted band size: 83 kDa

Observed band size: 83 kDa

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• WB

Lab

Western blot - Anti-HGF antibody [EPR12230] (AB178395)

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Western blot - Anti-HGF antibody [EPR12230] (ab178395) at 1/1000 dilution

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U-87 MG (Human glioblastoma-astrocytoma epithelial cell) whole cell lysates at 20 µg

Secondary

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Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 83 kDa

Observed band size: 83 kDa

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• WB

Lab

Western blot - Anti-HGF antibody [EPR12230] (AB178395)

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Western blot - Anti-HGF antibody [EPR12230] (ab178395) at 1/1000 dilution

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Human HGF (aa 32-728) recombinant protein

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 83 kDa

Observed band size: 60 kDa

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• WB

CiteAb

Western blot - Anti-HGF antibody [EPR12230] (AB178395)

HGF western blot using anti-HGF antibody [EPR12230] ab178395. Publication image and figure legend from Chen, H., Lin, C., et al., 2020, Cell Death Dis, PubMed 32041944.

ab178395 was used in this publication in western blot. This may not be the same as the application(s) guaranteed by Abcam. For a full list of applications guaranteed by Abcam for ab178395 please see the product overview.

The HGF/MET signalling pathway contributes to resistance to alectinib in ALK-positive lung cancer cells.a The concentrations of HGF in the H3122 and H2228 cell culture supernatants with or without alectinib (50 nmol/L) or crizotinib (100 nmol/L) administration were determined by ELISA. The data are presented as the mean ± SD, and the experiment was repeated three times. *p < 0.05; **p < 0.001. b H3122 and H2228 cells were treated with alectinib (50 nmol/L) or crizotinib (100 nmol/L) for the indicated amounts of time, and MET phosphorylation was measured by western blot. c, d H3122 and H2228 cells were transfected with HGF, and the expression of HGF and the phosphorylation levels of MET were determined by western blot. Vec negative control vector. e Cell proliferation in crizotinib-treated and alectinib-treated cells with or without HGF was examined by MTT assay; the IC50 values for crizotinib and alectinib were calculated. The data are presented as the mean ± SD, and the experiment was repeated three times. **p < 0.001. Cri crizotinib, Ale alectinib, Vec negative control vector. f H3122-AR1-6 cells are resistant to alectinib. The alectinib IC50 values of H3122 and H3122-AR cells were detected by MTT assay. The data are presented as the mean ± SD from three independent experiments. **p < 0.001 compared with H3122 cells. g Detection of HGF in the cell culture supernatants of H3122 and H3122-AR cells by ELISA. The data are presented as the mean ± SD from three independent experiments. Asterisks (*) indicates p < 0.05 compared with parental H3122 cells; **p < 0.001 compared with parental H3122 cells. h Immunoblot analysis of ALK, MET, and the phosphorylation of MET in H3122 and H3122-AR cell lines.

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