essentially as we described in (Tano et al., 2011 (link); Tano and Vazquez, 2011 (link)). Briefly, following cell lysis, solubilized proteins were separated in 10% acrylamide gels, electrotransferred to PVDF membranes and immunoblotted with the indicated primary antibody. After incubation with HRP-conjugated secondary antibodies, immunoreactive bands were visualized by ECL (Amersham, PA). Primary antibodies against IRE1α, BiP, CHOP, phospho-JNK1/2 (Thr183/Tyr185), total JNK1/2, phospho-STAT1 (Ser727) and total STAT1, were all from Cell Signaling (MA); the antibody against phospho-CAMKII (Thr286) was from Abcam (MA); the antibody against GAPDH was from Santa Cruz Biotechnology (Santa Cruz, CA) and the anti-TRPC3 antibody was from Alomone Labs (Jerusalem, Israel). In our hands, two different commercially available pan-CAMKII antibodies gave inconsistent, low quality results; thus, phospho-CAMKII levels were normalized by GAPDH.
Anti trpc3 antibody
Manufactured by Alomone
Sourced in Israel
The Anti-TRPC3 antibody is a research tool used to detect and quantify the expression of the TRPC3 protein, which is a member of the transient receptor potential (TRP) cation channel family. This antibody can be used in various techniques such as Western blotting, immunohistochemistry, and immunoprecipitation to study the localization and expression of TRPC3 in different cell types and tissues.
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Lab products found in correlation
Antibody against gapdh, by Santa Cruz Biotechnology (1 mentions)
Vectastain abc system, by Vector Laboratories (1 mentions)
Nis elements 3, by Nikon (1 mentions)
Alexa fluor 488 labelled secondary antibody, by Abcam (1 mentions)
Anti α sma antibody, by Abcam (1 mentions)
Anti tgf β1 antibody, by Abcam (1 mentions)
2 protocols using anti trpc3 antibody
1
Western Blot Immunodetection Protocol
2
Quantification of Myofibroblasts in Tissue Sections
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Tissue sections were blocked and incubated with an anti-TRPC3 antibody (1 : 100, Alomone Labs, Jerusalem, Israel) and an anti-TGFβ1 antibody (1 : 100, Abcam, Cambridge, UK) for 2 h at room temperature. The sections were incubated with a biotinylated secondary antibody for 30 minutes and developed with ABC complex (VECTASTAIN ABC System, Vector Labs, CA, USA). Immunofluorescence was performed with an anti-αSMA antibody (1 : 100, Abcam, Cambridge, UK) followed by an Alexa Fluor 488-labelled secondary antibody (Abcam, Cambridge, UK). Nuclei were identified by DAPI staining. To quantify fluorescence, the glass slides were examined under an inverted fluorescence microscope (Nikon TE2000-U; Olympus, Tokyo, Japan). The percentages of myofibroblast cells (αSMA+, green) among total cells were quantified using NIS-Elements 3.0 software (Nikon Instruments).
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