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Ab135813

Manufactured by Abcam
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Ab135813 is a primary antibody targeted against the protein of interest. It is suitable for use in various immunoassay techniques, such as Western blotting and immunohistochemistry. The antibody is purified and provided in a liquid formulation.

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Lab products found in correlation

A10042, by Thermo Fisher Scientific (3 mentions) Ab150129, by Abcam (3 mentions) Powerix70, by Olympus (2 mentions) Ab33586, by Abcam (1 mentions) Ab108624, by Abcam (1 mentions) Ab24590, by Abcam (1 mentions) Ab150107, by Abcam (1 mentions) Ab121893, by Abcam (1 mentions) Ab181604, by Abcam (1 mentions)

4 protocols using ab135813

1

Cytochrome P450 Antibody Validation

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ITE was a gift from Dr Jiasheng Song, (AhR Pharmaceuticals, Inc., Madison, WI, USA). Anti-CYP450 isoenzyme 1A1 antibody (ab126828), anti-CYP450 isoenzyme 1B1 antibody (ab33586), anti-CYP450 isoenzyme 1A2 antibody (ab56073), CYP3A4 antibody (ab135813), CYP3A5 antibody (ab108624), CYP2D6 antibody (ab62204), CYP2C9 antibody (ab150364) and CYP2E1 antibody (ab90564) were all purchased from Abcam (Cambridge, MA, USA).
Zhang X., Lu J., He B., Tang L., Liu X., Zhu D., Cao H., Wang Y, & Li L. (2016). A tryptophan derivative, ITE, enhances liver cell metabolic functions in vitro. International Journal of Molecular Medicine, 39(1), 101-112.
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2

Multicolor Immunofluorescence Staining

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Staining was performed following the protocol detailed in supplementary material File 1. The antibodies used in this study were goat anti-albumin (A80–129A, Bethyl), rabbit anti-CYP3A4 (ab135813, abcam), rabbit anti-Stabilin-2 (ab121893, abcam), mouse anti-PECAM-1 (ab24590, abcam), donkey anti-goat Alexa Fluor 488 (ab150129, abcam), donkey anti-rabbit Alexa Fluor 568 (A10042, Thermofisher), and donkey anti-mouse Alexa Fluor 647 (ab150107, abcam). Fluorescent images were captured using a confocal microscope (PowerIX70, Olympus).
Danoy M., Tauran Y., Poulain S., Jellali R., Bruce J., Leduc M., Le Gall M., Koui Y., Arakawa H., Gilard F., Gakiere B., Kato Y., Plessy C., Kido T., Miyajima A., Sakai Y, & Leclerc E. (2021). Investigation of the hepatic development in the coculture of hiPSCs-derived LSECs and HLCs in a fluidic microenvironment. APL Bioengineering, 5(2), 026104.
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3

Immunostaining of Liver Cell Markers

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Cells were fixed in 4% paraformaldehyde overnight at 4°C, washed and stored in PBS. Immunostaining was performed following the protocol previously described (Danoy et al., 2019) . The antibodies used were: Albumin (goat, A80-129A, Bethyl), CYP3A4 (rabbit, ab135813, abcam), anti-goat Alexa Fluor 488 (donkey, ab150129, abcam), and anti-rabbit Alexa Fluor 568 (donkey, A10042, Thermofisher). All fluorescent images were taken using a confocal microscope (PowerIX70, Olympus).
Danoy M., Poulain S., Lereau-Bernier M., Kato S., Scheidecker B., Kido T., Miyajima A., Sakai Y., Plessy C, & Leclerc E. (2020). Characterization of liver zonation-like transcriptomic patterns in HLCs derived from hiPSCs in a microfluidic biochip environment. Biotechnology progress, 36(5).
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4

Immunostaining of Hepatic Markers

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Fixation with 4% paraformaldehyde was performed overnight on all prepared samples at 4°C. Samples were then washed and stored at 4°C until the immunostaining started. Permeabilization and blocking of the samples were done with 0.1% Triton X-100 (plusone) and a pork gelatin buffer for 15 minutes and 2 hours respectively. Samples were then incubated with first antibodies (CYP3A4, HNF4a, ALB, AFP, diluted at 1/100, 1/1000, 1/1000 and 1/1000 respectively) overnight at 4°C followed with secondary antibodies diluted at 1/200 for 3 hours at room temperature.
After incubation with antibodies, samples were washed 3 times with PBS. Incubation with DAPI was performed for 15 min, the samples were then washed twice with PBS and kept in PBS for confocal imaging. The antibodies used were Albumin (goat, A80-129A, Bethyl), AFP (goat, sc-8109, Santa Cruz), CYP3A4 (Rabbit, ab135813, abcam), HNF-4Alpha (Rabbit, ab181604, abcam), anti-goat Alexa Fluor 488 (Donkey, ab150129, abcam) and anti-rabbit Alexa Fluor 568 (Donkey, A10042, Thermofisher).
Tauran Y., Poulain S., Lereau-Bernier M., Danoy M., Shinohara M., Segard B.D., Kato S., Kido T., Miyajima A., Sakai Y., Plessy C, & Leclerc E. (2019). Analysis of the transcription factors and their regulatory roles during a step-by-step differentiation of induced pluripotent stem cells into hepatocyte-like cells. Molecular omics, 15(6).
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