rabbit anti p gp  (Proteintech)

Journal: bioRxiv

Article Title: P-glycoprotein exofection between fetal and maternal cells as a mechanism of intercellular material transfer at the feto maternal interface

doi: 10.64898/2026.01.04.697556

Figure Lengend Snippet: (A) Immunohistochemical localization of P-gp in human fetal membrane tissues, showing the amnion epithelial layer (AEC), underlying chorionic trophoblast cells (CTCs), and maternal decidua. Representative control (left) and P-gp–stained sections (right) are shown at 10× magnification. (B) Schematic of the transwell configuration used for fetal membrane explant studies, with the fetal side oriented apically (left) or the maternal decidual side oriented apically (right). (C) Tacrolimus transport across fetal membrane explants quantified by LC–MS/MS, demonstrating directional efflux from the fetal to the maternal compartment. (D) Transwell systems using isolated CTCs (left) and DECs (right) to assess cell-specific transporter activity. (E) Tacrolimus efflux measured in CTC and DEC monolayers, showing higher efflux capacity in CTCs. (F) Relative ABCB1 mRNA expression in CTCs and DECs determined by qPCR. (G) Flow cytometric quantification of intracellular P-gp protein levels in DECs and CTCs using PE-conjugated antibody staining. Data are presented as mean ± SEM. Statistical analysis was performed using Student’s t-test. p < 0.05, p < 0.01, * p < 0.001.

Article Snippet: Secondary antibody for P-gp anti-rabbit (1:10,000, Amersham, Cat# NA934VS), and for the CD markers, the secondary antibodies were provided by the SBI kit EXOAB-kit-1 at a 1:5000 dilution.

Techniques: Immunohistochemical staining, Membrane, Control, Staining, Liquid Chromatography with Mass Spectroscopy, Isolation, Activity Assay, Expressing

Journal: bioRxiv

Article Title: P-glycoprotein exofection between fetal and maternal cells as a mechanism of intercellular material transfer at the feto maternal interface

doi: 10.64898/2026.01.04.697556

Figure Lengend Snippet: (A) Multiplex cytokine profiling of IL-6, IL-8, GM-CSF, and TNF in DECs and CTCs following LPS exposure, showing robust inflammatory cytokine release in DECs and minimal induction in CTCs. (B–C) Representative immunofluorescence images of P-gp (cyan) and DAPI (blue) in DECs (B) and CTCs (C) under control and LPS-treated conditions, with corresponding merged images. Bar graphs show quantification of P-gp mean fluorescence intensity. (D–E) Calcein-AM dye accumulation in DECs (D) and CTCs (E) under control, LPS, and verapamil-treated conditions, assessing P-gp–dependent efflux activity. Increased fluorescence denotes reduced efflux. Data are presented as mean ± SEM. Statistical analyses were performed using Student’s t-test or one-way ANOVA. p < 0.05, p < 0.01, * p < 0.001.

Article Snippet: Secondary antibody for P-gp anti-rabbit (1:10,000, Amersham, Cat# NA934VS), and for the CD markers, the secondary antibodies were provided by the SBI kit EXOAB-kit-1 at a 1:5000 dilution.

Techniques: Multiplex Assay, Immunofluorescence, Control, Fluorescence, Activity Assay

Journal: bioRxiv

Article Title: P-glycoprotein exofection between fetal and maternal cells as a mechanism of intercellular material transfer at the feto maternal interface

doi: 10.64898/2026.01.04.697556

Figure Lengend Snippet: (A) Schematic overview of the EV isolation and purification workflow from CTC-conditioned media using tangential flow filtration (TFF). (B) Comparative proteomic analysis of transporter proteins in EVs derived from DECs and CTCs under control and LPS-stimulated conditions, highlighting enrichment of transporter families in CTC-EVs. (C) Nanoparticle tracking analysis (Zetaview) showing EV size distribution and particle concentration. (D) Cryo-electron microscopy image demonstrating intact lipid bilayer structures and morphological heterogeneity of EVs. (E) Super-resolution Nanoimager microscopy showing surface expression of canonical EV tetraspanins (CD9, CD63, CD81). (F) Western blot analysis confirming the presence of EV markers (CD9, CD63, CD81), HSP70, purity marker Calnexin, and P-gp cargo in isolated EVs. (G) ELISA-based quantification of P-gp abundance in EV preparations. (H) Functional annotation of proteins identified in CTC-derived EVs, demonstrating enrichment of pathways involved in membrane fusion, vesicle trafficking, receptor-mediated interactions, lysosomal processes, cellular adhesion, and molecular transport.

Article Snippet: Secondary antibody for P-gp anti-rabbit (1:10,000, Amersham, Cat# NA934VS), and for the CD markers, the secondary antibodies were provided by the SBI kit EXOAB-kit-1 at a 1:5000 dilution.

Techniques: Isolation, Purification, Filtration, Derivative Assay, Control, Concentration Assay, Cryo-Electron Microscopy, Microscopy, Expressing, Western Blot, Marker, Enzyme-linked Immunosorbent Assay, Functional Assay, Membrane

Journal: bioRxiv

Article Title: P-glycoprotein exofection between fetal and maternal cells as a mechanism of intercellular material transfer at the feto maternal interface

doi: 10.64898/2026.01.04.697556

Figure Lengend Snippet: (A) Immunofluorescence analysis of P-gp expression in DECs under four treatment conditions: Control, LPS, EVs, and LPS + EVs. Nuclei were stained with DAPI (blue) and P-gp with a FITC-conjugated antibody (green). Images captured at 20× magnification. Quantification of P-gp mean fluorescence intensity is shown in the accompanying bar graph. (B) Functional assessment of P-gp activity using a calcein-AM efflux assay across the same conditions. Verapamil was used as a positive control to confirm efflux specificity. (C) Generation of transporter-deficient DECs using ABCB1-targeted siRNA. Knockdown efficiency was validated by qPCR. (D) Loss of P-gp–mediated efflux in knockdown (KD) cells confirmed by increased intracellular calcein accumulation. (E) Rescue of P-gp protein expression in P-gp–deficient DECs following treatment with CTC-derived EVs. Groups include siRNA control, P-gp KD, and P-gp KD + EVs. Representative immunofluorescence images and quantified fluorescence intensities illustrate EV-mediated restoration of P-gp protein. (F) Functional restoration of P-gp activity in KD cells following EV treatment, measured using a dye efflux assay. EV treatment significantly reduced dye accumulation relative to untreated KD cells, demonstrating recovery of transporter function. Statistical significance was determined using Student’s t -test or one-way ANOVA. Data are presented as Mean ± SEM. Significance is denoted as * for p < 0.05, ** for p < 0.01 and *** for p < 0.001

Article Snippet: Secondary antibody for P-gp anti-rabbit (1:10,000, Amersham, Cat# NA934VS), and for the CD markers, the secondary antibodies were provided by the SBI kit EXOAB-kit-1 at a 1:5000 dilution.

Techniques: Immunofluorescence, Expressing, Control, Staining, Fluorescence, Functional Assay, Activity Assay, Positive Control, Knockdown, Derivative Assay

Journal: bioRxiv

Article Title: P-glycoprotein exofection between fetal and maternal cells as a mechanism of intercellular material transfer at the feto maternal interface

doi: 10.64898/2026.01.04.697556

Figure Lengend Snippet: (A) Schematic overview of the experimental workflow. Pregnant mice were assigned to three groups: wild-type (WT), P-gp knockout (P-gp KO), and P-gp KO treated with CTC-derived exosomes (KO+EV). EVs (1×10¹ particles per dose) were administered on gestational day E14, followed by tacrolimus dosing (2 mg/kg). Plasma and tissues were collected at specified timepoints for pharmacokinetic analysis. (B) Tacrolimus exposure profiles plotted over time for each group. The area under the concentration–time curve (AUC ₀ – ₅ h) was quantified by LC-MS and compared across WT, KO, and KO+EV animals. (C) Tacrolimus plasma concentrations at the 0.5-hour timepoint, illustrating impaired clearance in KO mice and partial restoration following EV treatment. (D) Maximum plasma concentration (Cmax) of tacrolimus in each group. (E) Comparative pharmacokinetic parameters—including elimination rate constant (k_el), half-life (t ₁ / ₂ ), Tmax, and mean residence time (MRT)—demonstrating EV-mediated improvement of tacrolimus disposition in P-gp–deficient mice. (F–G) Tissue-specific accumulation of tacrolimus in the placenta (F) and lung (G). EV treatment significantly reduced aberrant drug accumulation in KO animals, indicating partial restoration of P-gp–mediated efflux in vivo. Statistical analyses were performed using Student’s t -test or one- or two-way ANOVA where appropriate. Data are presented as Mean ± SEM. Statistical significance is denoted as * for p < 0.05, ** for p < 0.01 and *** for p < 0.001

Article Snippet: Secondary antibody for P-gp anti-rabbit (1:10,000, Amersham, Cat# NA934VS), and for the CD markers, the secondary antibodies were provided by the SBI kit EXOAB-kit-1 at a 1:5000 dilution.

Techniques: Knock-Out, Derivative Assay, Clinical Proteomics, Concentration Assay, Liquid Chromatography with Mass Spectroscopy, In Vivo