- 702069 - Invitrogen Antibodies APLNR antibody

Submitted references The Apelin-Apelin Receptor Axis Triggers Cholangiocyte Proliferation and Liver Fibrosis During Mouse Models of Cholestasis.

Chen L, Zhou T, White T, O'Brien A, Chakraborty S, Liangpunsakul S, Yang Z, Kennedy L, Saxena R, Wu C, Meng F, Huang Q, Francis H, Alpini G, Glaser S
Hepatology (Baltimore, Md.) 2021 Jun;73(6):2411-2428

Spatiotemporal extracellular matrix modeling for in situ cell niche studies.

Olesen K, Rodin S, Mak WC, Felldin U, Österholm C, Tilevik A, Grinnemo KH
Stem cells (Dayton, Ohio) 2021 Dec;39(12):1751-1765

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis was performed on membrane enriched extract/ tissue extract (30 µg lysate) of HepG2 (Lane 1), NTERA-2 (Lane 2), HEL.92.1.7 (Lane 3), K562 (Lane 4), and mouse liver (Lane 5). The blots were probed with Anti-APLNR Recombinant Rabbit Monoclonal Antibody (Product # 702069, 1-2 µg/mL) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.4 µg/mL, 1:2500 dilution). A 50 kDa band corresponding to APLNR protein was observed. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 4-12% Bis-Tris gel (Product # NP0321BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary Antibody following blocking with 5% skimmed milk. Chemiluminescent detection was performed using Pierce™ ECL Western blotting Substrate (Product # 32106).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis was performed on membrane enriched extract/ tissue extract (30 µg lysate) of HepG2 (Lane 1), NTERA-2 (Lane 2), HEL.92.1.7 (Lane 3), K562 (Lane 4), and mouse liver (Lane 5). The blots were probed with Anti-APLNR Recombinant Rabbit Monoclonal Antibody (Product # 702069, 1-2 µg/mL) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.4 µg/mL, 1:2500 dilution). A 50 kDa band corresponding to APLNR protein was observed. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 4-12% Bis-Tris gel (Product # NP0321BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary Antibody following blocking with 5% skimmed milk. Chemiluminescent detection was performed using Pierce™ ECL Western blotting Substrate (Product # 32106).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence was performed on fixed and permeabilized HeLa cells for detection of endogenous APLNR using Anti-APLNR Recombinant Rabbit Monoclonal Antibody (Product # 702069, 2 µg/mL) and labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034, 1:2000). Panel a) shows representative cells that were stained for detection and localization of APLNR protein (green), Panel b) is stained for nuclei (blue) using SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). Panel c) represents cytoskeletal F-actin staining using Alexa Fluor® 555 Rhodamine Phalloidin (Product # R415, 1:300). Panel d) is a composite image of Panels a, b and c clearly demonstrating membrane localization of APLNR. Panel e) represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: 1 FIG. APJ-mediated APLN expression is increased in cholangiocytes during cholestasis. (A) Immunofluorescence of APLN (costained with CK-19) in liver sections from patients with PSC (n = 7). Upper three panels, original magnification, x40; scale bar, 50 mum. Lower panel, original magnification, x40 zoom5; scale bar, 10 mum (red CK-19, green APLN, blue 4',6-diamidino-2-phenylindole [DAPI]). (B) Immunofluorescence of APJ (costained with CK-19) in liver sections from patients with PSC (n = 7). Upper three panels, original magnification, x40; scale bar, 50 mum. Lower panel, original magnification, x40 zoom5; scale bar, 10 mum (red CK-19, green APJ, blue DAPI). (C) The mRNA expression of APLN and APLNR in cholangiocytes from patients with PSC (hPSCL; mean +- SD, n = 3), * P < 0.05 versus HIBEpiCs. (D) APLN levels in serum of patients with PSC (mean +- SD, n = 26), * P < 0.05 versus control. (E) APLN levels in BDL mouse serum and cholangiocyte supernatant (mean +- SD, n = 3), * P < 0.05 versus WT, # P < 0.05 versus BDL. (F) APLN levels in Mdr2 -/- mouse serum (mean +- SD, n = 3), * P < 0.05 versus WT, # P < 0.05 versus Mdr2 -/- .

702069

Antibody data