This was achieved using 1:1000 polyclonal antibodies against phosphorylated insulin receptor beta (p-IRβ; phospho-Y1361), IR β (C18C4) (Abcam, Cambridge, MA, USA), phosphorylated Akt (p-Ser473 Akt), Akt, α-FASN (Cell Signaling, Danvers, MA, USA), custom-made rabbit polyclonal Ab2456 against the mouse CEACAM1 extracellular domain, as titrated [20 (link)], and custom-made rat CEACAM1 (αP3[Y488]) and phosphorylated CEACAM1 (α-p-CEACAM1) mouse antibodies (Bethyl Laboratories, Montgomery, TX, USA), as titrated [16 (link), 21 (link)]. α-Actin, α-GAPDH and α-tubulin antibodies (Santa Cruz) were used at 1:5000 dilution for normalisation. Blots were incubated with horseradish peroxidase-conjugated donkey anti-rabbit IgG antibody, as per manufacturer’s titration, for analysis of transfected, knockout or siRNA-knockdown cells (GE Healthcare Life Sciences, Amersham, Sunnyvale, CA, USA), followed by ECL.
Horseradish peroxidase conjugated donkey anti rabbit igg antibody
Manufactured by GE Healthcare
Sourced in United States
Horseradish peroxidase-conjugated donkey anti-rabbit IgG antibody is a laboratory reagent used for immunodetection applications. It consists of a donkey-derived polyclonal antibody that specifically binds to rabbit immunoglobulin G (IgG) molecules, with the antibody conjugated to the enzyme horseradish peroxidase.
Automatically generated - may contain errors
Lab products found in correlation
Enhanced chemi luminescence (ecl), by Cytiva (2 mentions)
α actin, by Santa Cruz Biotechnology (1 mentions)
α tubulin antibody, by Santa Cruz Biotechnology (1 mentions)
α gapdh, by Santa Cruz Biotechnology (1 mentions)
α fasn, by Cell Signaling Technology (1 mentions)
Phospho smad2 ser465 467, by Cell Signaling Technology (1 mentions)
Phospho smad3 ser423 425, by Cell Signaling Technology (1 mentions)
Igf 1, by US Biological (1 mentions)
Anti p44 42 erk antibody, by Cell Signaling Technology (1 mentions)
α tubulin monoclonal antibody, by Cell Signaling Technology (1 mentions)
Anti mouse igg antibody, by GE Healthcare (1 mentions)
Phospho akt ser473, by Cell Signaling Technology (1 mentions)
α pcc1, by Fortis Life Sciences (1 mentions)
Polyclonal antibody, by Cell Signaling Technology (1 mentions)
Horseradish peroxidase conjugated, by Jackson ImmunoResearch (1 mentions)
5 protocols using horseradish peroxidase conjugated donkey anti rabbit igg antibody
1
Western Blot Analysis of Insulin Signaling
2
Phospho-Smad2/3 Immunoblotting Analysis
3
IGF-IR and ERK1/2 Signaling Analysis
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Type 1 insulin-like growth factor receptor levels were analyzed by Western blotting performed as described in detail elsewhere (12 (link)) and using a rabbit anti-IGF-IR antiserum (C-20, Santa Cruz Biotechnology, Dallas, TX, United States) and a horseradish peroxidase-conjugated donkey anti rabbit IgG antibody (GE Healthcare Life Sciences, Pittsburgh, PA, United States). To normalize for loading, the membranes were stripped and re-probed with a monoclonal anti-tyrosine tubulin antibody (Sigma-Aldrich, St. Louis, MO, United States). To analyze ERK1 and 2 phosphorylation in response to IGF-I, tumor cells, cultured overnight in serum-free medium, were stimulated with 100 ng/ml IGF-I (US Biological, Salem, MA, United States) for 10 min, lysed on ice in the presence of phosphatase inhibitors and the cell lysates separated on 8.5% SDS-polyacrylamide gels. The blots were probed, first with an anti-phospho-p44/42 ERK (Thr202/Tyr204) antibody and then with an anti p44/42 ERK antibody (Cell Signaling, Whitby, ON, Canada).
4
CEACAM1 and Insulin Signaling Analysis
5
Immunoblotting and Protein Interactions
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Tissues and cells were lysed and proteins analyzed by SDS-PAGE followed by immunoprobing with polyclonal antibodies (Cell signaling, Danvers, MA, USA) against: phospho-Akt (Ser473), Akt, phospho-p44/42 MAPK (Thr202/Tyr204), p44/42 MAPK, phospho-eNOS (Ser1177), eNOS, phospho-Smad2Ser465/467, Smad2, phospho-Smad3 (Ser423/425), Smad3, phospho-Stat3 (Tyr705), Stat3, phospho-NF-ϏB (Ser536), NF-ϏB, phospho-PKCζ (phospho-Thr410/403), PKCζ, phospho-VEGFR2 (Try1175), VEGF-R2, VCAM1 and Tubulin. A custom-made rabbit polyclonal antibody (Ab 3759) were used against mouse CEACAM1 extracellular domain and phospho-CEACAM1 (α-pCC1) (Bethyl Laboratories, Montgomery, TX, USA). Blots were incubated with horseradish peroxidase-conjugated donkey anti-rabbit IgG antibody (GE Healthcare Life Sciences, Amersham, Marlborough, MA, USA) and proteins were visualized using ECL (Amersham). polyclonal antibodies against SHP2, and Shc (Cell signaling) were used in co-immunoprecipitation experiments. These membranes were incubated with light chain specific horseradish peroxidase-conjugated IgG monoclonal mouse anti-rabbit (211-032- 171, Jackson Immuno-Research laboratories) to prevent detection of the antibody heavy chain.
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