To investigate the activation of CpG, we measured the expression of Toll-like receptor 9 (TLR9) and myeloid differentiation primary response 88 (Myd88) by using western blotting [22 (link)]. The pPCFN-CpG, pVAX1-PCFN, and empty vector pVAX1 were transfected into the RAW264.7 using lipofectamine 2000. After 36 h, the samples derived from RAW264.7 were collected and lysed in RIPA buffer, separated by electrophoresis on 12% SDS-PAGE gels and transferred to nitrocellulose (GE Amersham Biosciences, Piscataway, NJ, USA). Proteins were detected by western blotting using primary antibodies (Abs) (Mouse monoclonal Abs against GAPDH, Myd88, TLR9) at a concentration of 1/1000 (Santa Cruz Biotechnology, Santa Cruz, CA, USA) and were incubated at 4 °C overnight. Labeling of the first Abs was detected using goat anti-mouse Abs conjugated to HRP (Santa Cruz Biotechnology) and detected by using ECL reagents.
Mouse monoclonal abs against gapdh
Manufactured by Santa Cruz Biotechnology
Sourced in Canada
Mouse monoclonal antibodies against GAPDH. GAPDH is a widely-used reference protein for immunoblotting and other applications.
Automatically generated - may contain errors
Lab products found in correlation
Goat polyclonal abs against il, by Santa Cruz Biotechnology (2 mentions)
Rabbit monoclonal ab against phosphor nf κb p65, by Cell Signaling Technology (2 mentions)
Hospho irak4, by Cell Signaling Technology (2 mentions)
K48 linked polyub chains, by Cell Signaling Technology (2 mentions)
K63 linked polyub chains, by Cell Signaling Technology (2 mentions)
Ubiquitin, by Cell Signaling Technology (2 mentions)
A20 tnfaip3, by Cell Signaling Technology (2 mentions)
3 protocols using mouse monoclonal abs against gapdh
1
Investigating CpG Activation via TLR9 and MyD88
2
Quantification of NF-κB Pathway Activation
3
Quantification of NF-κB Pathway Activation
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Mouse monoclonal Abs against GAPDH and IL-6 and goat polyclonal Abs against IL-1β and TNF-α were obtained from Santa Cruz Biotechnology, Inc (Santa Cruz, CA). Rabbit monoclonal Ab against phosphor-NF-κB p65, hospho-IRAK4, A20/TNFAIP3, Ubiquitin, K48-linked polyUb chains, K63-linked polyUb chains and IRAK4 were purchased from Cell Signaling Technology (Cambridge, MA). The samples derived from cells and lung homogenates were lysed in RIPA buffer, separated by electrophoresis on 12% SDS-PAGE gels and transferred to nitrocellulose (GE Amersham Biosciences, Piscataway, NJ). Proteins were detected by western blotting using primary Abs at a concentration of 1/200 (Santa Cruz Biotechnology) or 1/1000 (Cell Signaling Technology) and were incubated overnight. Labeling of the first Abs was detected using relevant secondary Abs conjugated to HRP (Santa Cruz Biotechnology, Santa Cruz, CA) and detected using ECL reagents.
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