- PA5-78690 - Invitrogen Antibodies ABCB11 antibody

Li Z, Chiang YP, He M, Worgall TS, Zhou H, Jiang XC
iScience 2021 Dec 17;24(12):103449

Li H, Xi Y, Xin X, Tian H, Hu Y
Nutrition & metabolism 2020;17:34

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis of BSEP in Lane 1: rat liver tissue lysate, Lane 2: mouse liver tissue lysate, Lane 3: SMMC whole cell lysate using 50 µg (reducing conditions) per well. Electrophoresis was performed on 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours and protein was transferred to a nitrocellulose membrane at 150mA for 50-90 minutes. Sample was blocked with 5% Non-fat Milk/TBS for 1.5 hours at room temperature, incubated with BSEP polyclonal antibody (Product # PA5-78690) at a dilution of 0.5 µg/mL (overnight at 4°C), followed by goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:10,000. Signal development was performed using a chemiluminescence (ECL) kit.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of ABCB11 in, Lane 1: human HCCT tissue lysates, Lane 2: rat liver tissue lysates, Lane 3: mouse liver tissue lysates. Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30 µg of sample under reducing conditions. After Electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. The membrane was blocked with 5% non-fat milk/TBS for 1. 5 hour at RT. The membrane was incubated with BSEP Polyclonal Antibody (Product # PA5-78690) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0. 1% Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5,000 for 1. 5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit. A specific band was detected for ABCB11 at approximately 146 kDa. The expected band size for ABCB11 is at 146 kDa.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunohistochemistry analysis of BSEP on paraffin-embedded rat liver tissue. Antigen retrieval was performed using citrate buffer (pH6, epitope retrieval solution) for 20 mins. Sample was blocked using 10% goat serum, incubated with BSEP polyclonal antibody (Product# PA5-78690) with a dilution of 1 µg/mL (overnight at 4°C), and followed by biotinylated goat anti-rabbit IgG (30 minutes at 37°C). Development was performed using Streptavidin-Biotin-Complex (SABC) with DAB chromogen method.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemistry analysis of BSEP on paraffin-embedded mouse liver tissue. Antigen retrieval was performed using citrate buffer (pH6, epitope retrieval solution) for 20 mins. Sample was blocked using 10% goat serum, incubated with BSEP polyclonal antibody (Product# PA5-78690) with a dilution of 1 µg/mL (overnight at 4°C), and followed by biotinylated goat anti-rabbit IgG (30 minutes at 37°C). Development was performed using Streptavidin-Biotin-Complex (SABC) with DAB chromogen method.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemistry analysis of BSEP on paraffin-embedded human liver cancer tissue. Antigen retrieval was performed using citrate buffer (pH6, epitope retrieval solution) for 20 mins. Sample was blocked using 10% goat serum, incubated with BSEP polyclonal antibody (Product# PA5-78690) with a dilution of 1 µg/mL (overnight at 4°C), and followed by biotinylated goat anti-rabbit IgG (30 minutes at 37°C). Development was performed using Streptavidin-Biotin-Complex (SABC) with DAB chromogen method.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemical analysis of ABCB11 in paraffin-embedded section of mouse liver tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution ) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/mL rabbit anti-ABCB11 antibody (Product # PA5-78690) overnight at 4°C. Cy3 Conjugated Goat Anti-Rabbit IgG was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Flow Cytometry of BSEP in A431 cells (blue line), isotype control rabbit IgG (green line) and unlabeled (red line). Samples were blocked with 10% goat serum, incubated with BSEP Polyclonal Antibody (Product # PA5-78690) at a dilution of 1 μg (per 1x10^6 cells), followed by DyLight®488 conjugated goat anti-rabbit IgG (for 30 minutes at 20°C) using 5-10 μg (per 1x10^6 cells) dilution.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Fig. 11 Changes in FGF15, FGFR4, KLB, CYP7A1, NTCP, and BSEP protein content in liver tissues among the different experimental groups. a Bar chart representing the relative CYP7A1 protein expression level in liver tissue, b relative NTCP protein expression level, c relative BSEP protein expression level, d relative FGF15 protein expression level, e relative FGFR4 protein expression level, and f relative KLB protein expression level in liver tissue. g Representative autoradiograph image of western blotting with the different antibodies. ** P < 0.01, vs control group; ## P < 0.01, # P < 0.05, vs HFD group. Data are presented as mean +- standard deviation, n = 3. C, control; HFD, high-fat diet; Gyp, gypenosides; OCA, obeticholic acid

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 2 SMS1 deficiency-mediated hepatocyte impairment Liver sections from 6-month-old WT, Sms2 KO, and Sms1 / Sms2 KO female mice were utilized. (A) Na + -taurocholate co-transporting polypeptide (NTCP) immunostaining for hepatocyte basal membrane. (B) Bile salt export pump (BSEP) immunostaining for hepatocyte apical membrane. The pictures are the representatives of four mice/group. (C) Plasma was collected from WT, Sms2 KO, and Sms1 / Sms2 KO mice, and jaundice was observed. (D) Fluorogram of plasma bilirubin (conjugated) levels. (E) Fluorogram of plasma total bile acid levels. (F and G) Fluorogram of plasma alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels. Data are represented as mean +- SD, n = 4-6, *p < 0.01.

PA5-78690