Following antibodies were used: anti-GAPDH (HRP-60004, Proteintech), anti-6∗His (ab18184, Abcam), anti-GFP (11814460001, Roche), anti-Myc (2278S, Cell Signaling Technology), anti-ACBD3 (H00064746-B01P, Abnova), anti-P-PKA-substrate(9621S, Cell Signaling Technology), anti-Golgin97 (13192, Cell Signaling Technology), anti-mCherry (ab167453, Abcam), anti-PKA-Cα (#4782, Cell Signaling Technology), anti-PKA-RIIα (612242, BD), anti-γ-COP (sc-393615, Santa Cruz), anti-GM130 (610822, BD Bioscience), anti-SNAP tag (P9310S, NEB), anti-Clathrin HC (ab21679, Abcam), anti-ARF1 (NB300-505, Novus), anti-β-Actin (3700S, Cell Signaling), and anti-α-Tubulin (3873S, Cell Signaling). Anti-Rabbit Alexa Fluor 488 (A21441), Alexa Fluor 568 (A10042), Alexa Fluor 647 (A21245), and anti-Mouse Alexa Fluor 488 (A21200), Alexa Fluor 568 (A10037), and Alexa Fluor 647 (A21236) for immunofluorescence staining were obtained from ThermoFisher. D/D Solubilizer (635054) was purchased from Clontech. Forskolin (F6886) was purchased from Sigma.
Alexa fluor 568 a10037
Manufactured by Thermo Fisher Scientific
Alexa Fluor 568 (A10037) is a fluorescent dye produced by Thermo Fisher Scientific. It is a synthetic dye that emits fluorescence at a wavelength of approximately 568 nanometers when excited by light at a suitable wavelength.
Automatically generated - may contain errors
Lab products found in correlation
Ab21679, by Abcam (1 mentions)
Ab167453, by Abcam (1 mentions)
Ab18184, by Abcam (1 mentions)
Anti mcherry, by Abcam (1 mentions)
Anti golgin97, by Cell Signaling Technology (1 mentions)
Anti snap tag, by New England Biolabs (1 mentions)
Alexa fluor 647 a21236, by Thermo Fisher Scientific (1 mentions)
Anti myc, by Cell Signaling Technology (1 mentions)
Anti pka c α, by Cell Signaling Technology (1 mentions)
Forskolin f6886, by Merck Group (1 mentions)
Anti α tubulin, by Cell Signaling Technology (1 mentions)
D d solubilizer, by Takara Bio (1 mentions)
Anti β actin, by Cell Signaling Technology (1 mentions)
Anti gm130, by BD (1 mentions)
Immobilon p pvdf membrane, by Merck Group (1 mentions)
Ha 11mms 101r, by Fortrea (1 mentions)
Immobilon western chemiluminescent hrp substrate, by Merck Group (1 mentions)
2 protocols using alexa fluor 568 a10037
1
Immunofluorescence Staining of Cellular Proteins
2
Western Blotting Protocol for Wnt Signaling
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Western blot method was done accordingly to the previously published protocol [28 ]. In brief, cells were washed with PBS and subsequently lysed in the sample buffer (2% SDS, 10% glycerol, 5% β-mercaptoethanol, 0.002% bromphenol blue and 0.06 M Tris HCl, pH 6.8), sonicated and thermally denatured. Electrophoretically separated proteins, in the SDS-PAGE gels, were transferred onto Immobilon-P PVDF Membrane (IPVH00010, Millipore) and detected by primary and corresponding HRP-conjugated secondary antibodies on Fusion SL imaging system (Vibler), using Immobilon Western Chemiluminescent HRP Substrate (Merck, WBKLS0500). Western blot signals were quantified using ImageJ software. Antibodies are listed in the Table 2 . Western blot membranes are presented together with molecular mass markers in the Additional file 1 .
Antibodies
| Antibody | Manufacturer | Dilution and application | Reference |
|---|---|---|---|
| LRP6 (C47E12) | cs-3395, Cell Signaling Technology | 1:1000, WB | [28 ] |
| LRP6 (Phospho-S1490) | cs-2568, Cell Signaling Technology | 1:1000, WB | [28 ] |
| β-actin | CS-4970, Cell Signaling Technology | 1:3000, WB | [26 (link)] |
| DVL-2 | CS-3216, Cell Signaling Technology | 1:1000, WB | [26 (link)] |
| DVL-3 | CS-3218, Cell Signaling Technology | 1:1000, WB | [26 (link)] |
| HA-11 | MMS-101R, Covance | 1:2000 WB; 1:500, IF | [28 ] |
| a-mouse IgG HRP | A4416 | 1:4000 WB | – |
| a-rabbit IgG HRP | A0545 | 1:4000 WB | – |
| a-mouse Alexa Fluor™568 | A10037, Thermo Fisher Scientific | 1:600 IF | – |
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