PA5-25614

antibody from Invitrogen Antibodies

Targeting: SLC10A1 NTCP

Western blot (Supportive data in Antibodypedia)

Flow cytometry (Supportive data in Antibodypedia)

Other assay (Supportive data in Antibodypedia)

Antibody Data

Product number

PA5-25614

Provider

Invitrogen Antibodies

Product name

NTCP Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Synthetic peptide

Reactivity

Human

Host

Rabbit

Isotype

IgG

Vial size

200 µL

Concentration

0.5 mg/mL

Storage

Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.

References

Submitted references

Robust in vitro assay for analyzing the neutralization activity of serum specimens against hepatitis B virus.

Zhang YL, Gao Y, Cao JL, Zhao JH, Zhang TY, Yang CL, Xiong HL, Wang YB, Ou SH, Cheng T, Chen CR, Yuan Q, Xia NS
Emerging microbes & infections 2019;8(1):724-733

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis of NTCP in various lysates. Samples were incubated with NTCP polyclonal antibody (Product # PA5-25614) using a dilution of 1:1,000 followed by Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at a dilution of 1:10,000. Lysates/proteins: 20 µg per lane. Lane 1: HT-1080 whole cell lysate; Lane 2: HepG2 whole cell lysate; Lane 3: K562 whole cell lysate; Lane 4: Caco2 whole cell lysate. Predicted band size: 38 kDa. Blocking/Dilution buffer: 5% NFDM/TBST.

Flow cytometry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Flow cytometry analysis of K562 cells using a SLC10A1 polyclonal antibody (Product # PA5-25614) (right) compared to a negative control cell (left) at a dilution of 1:10-50, followed by a FITC-conjugated goat anti-rabbit antibody

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Flow cytometry of (overlay histogram) of NTCP in HepG2 cells (green line). Samples were incubated with NTCP polyclonal antibody (Product # PA5-25614) using a dilution of 1:25 dilution for 60 min at 37°C followed by Goat-Anti-Rabbit IgG, DyLight® 488 Conjugated Highly Cross-Adsorbed at 1:200 dilution for 40 min at 37°C. The cells were fixed with 2% paraformaldehyde 10 min. The cells were then incubated in 2% bovine serum albumin to block non-specific protein-protein interactions followed by the primary antibody. Isotype control antibody (blue line) was rabbit IgG1 (1 μg/1x10^6 cells) used under the same conditions. Acquisition of >10, 000 events was performed.

Other assay

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Figure 1. Establishment and characterization of NTCP-reconstituted HepG2 cells for in vitro HBV infection. (A) A schematic diagram depicting the dox-inducible NTCP expression cassette. The NTCP-IRES-mCherry cassette was cloned into the pLenti-CMVTRE3G-eGFP (Addgene 27570) vector between the BamHI and XbaI restriction sites. (B) Flow cytometric analyses of the dox-induced mCherry expression of the different HepG2-TetOn-NTCP cell lines. The upper panel showed the percentage of mCherry+ cell population, and the lower panel showed the mean fluorescence intensity (MFI) of mCherry+ cell population of various cell lines. (C) Flow cytometric analyses of the dox-induced PreS1-peptide (FITC conjugated) binding capability of the different HepG2-TetOn-NTCP cell lines. The upper panel presented the dose-dependent comparison on the FITC/mCherry double positive percentage of different cell lines, and the lower panel presented the FITC MFI of FITC+/mCherry+ cell populations of various cell lines when binding with myr-PreS1-FITC peptide at different dosage (500, 100 and 20 nM). (D) Western blots for NTCP expression in the different HepG2-TetOn-NTCP cell lines with or without dox induction. The NTCP was detected by a commercial rabbit polyclonal antibody against NTCP (Thermo Fisher Scientific, PA525614). (E, F) Detections of HBeAg (E) and HBsAg (F) in culture supernatants of different HepG2-TetOn-NTCP cell lines following infection with HBV at 1 GE/cell (upper panel) or 100 GE/cell (lower pa