- 39-3800 - Invitrogen Antibodies PODXL antibody

Submitted references 3D organoid-derived human glomeruli for personalised podocyte disease modelling and drug screening.

Hale LJ, Howden SE, Phipson B, Lonsdale A, Er PX, Ghobrial I, Hosawi S, Wilson S, Lawlor KT, Khan S, Oshlack A, Quinlan C, Lennon R, Little MH
Nature communications 2018 Dec 4;9(1):5167

Urinary podocyte-associated mRNA levels correlate with proximal tubule dysfunction in early diabetic nephropathy of type 2 diabetes mellitus.

Petrica L, Ursoniu S, Gadalean F, Vlad A, Gluhovschi G, Dumitrascu V, Vlad D, Gluhovschi C, Velciov S, Bob F, Matusz P, Milas O, Secara A, Simulescu A, Popescu R
Diabetology & metabolic syndrome 2017;9:31

Therapy with atorvastatin versus rosuvastatin reduces urinary podocytes, podocyte-associated molecules, and proximal tubule dysfunction biomarkers in patients with type 2 diabetes mellitus: a pilot study.

Vlad A, Vlad M, Petrica L, Ursoniu S, Gadalean F, Popescu R, Vlad D, Dumitrascu V, Gluhovschi G, Gluhovschi C, Velciov S, Bob F, Matusz P, Secara A, Simulescu A, Jianu DC
Renal failure 2017 Nov;39(1):112-119

Urinary Podocyte Loss Is Increased in Patients with Fabry Disease and Correlates with Clinical Severity of Fabry Nephropathy.

Fall B, Scott CR, Mauer M, Shankland S, Pippin J, Jefferson JA, Wallace E, Warnock D, Najafian B
PloS one 2016;11(12):e0168346

Bmi1 essentially mediates podocalyxin-enhanced Cisplatin chemoresistance in oral tongue squamous cell carcinoma.

Zhou Y, Zhang L, Pan H, Wang B, Yan F, Fang X, Munnee K, Tang Z
PloS one 2015;10(4):e0123208

Podocalyxin promotes cisplatin chemoresistance in osteosarcoma cells through phosphatidylinositide 3-kinase signaling.

Huang Z, Huang Y, He H, Ni J
Molecular medicine reports 2015 Sep;12(3):3916-3922

Podocalyxin promotes glioblastoma multiforme cell invasion and proliferation by inhibiting angiotensin-(1-7)/Mas signaling.

Liu B, Liu Y, Jiang Y
Oncology reports 2015 May;33(5):2583-91

Podocalyxin promotes glioblastoma multiforme cell invasion and proliferation via β-catenin signaling.

Liu Y, Yang L, Liu B, Jiang YG
PloS one 2014;9(10):e111343

Sperm‑associated antigen 9 promotes astrocytoma cell invasion through the upregulation of podocalyxin.

Jiang J, Liu Y, Fang W, Liu F
Molecular medicine reports 2014 Jul;10(1):417-22

Stimulatory effects of cardiotrophin 1 on atherosclerosis.

Konii H, Sato K, Kikuchi S, Okiyama H, Watanabe R, Hasegawa A, Yamamoto K, Itoh F, Hirano T, Watanabe T
Hypertension (Dallas, Tex. : 1979) 2013 Nov;62(5):942-50

Role of podocalyxin in astrocytoma: Clinicopathological and in vitro evidence.

Huang T, Jin X, He L, Zhang M, Wu J, Wang Y, Fang J
Oncology letters 2013 Nov;6(5):1390-1396

A dimeric PINK1-containing complex on depolarized mitochondria stimulates Parkin recruitment.

Okatsu K, Uno M, Koyano F, Go E, Kimura M, Oka T, Tanaka K, Matsuda N
The Journal of biological chemistry 2013 Dec 20;288(51):36372-84

Podocalyxin regulates astrocytoma cell invasion and survival against temozolomide.

Wu H, Yang L, Liao D, Chen Y, Wang W, Fang J
Experimental and therapeutic medicine 2013 Apr;5(4):1025-1029

PINCH1 is transcriptional regulator in podocytes that interacts with WT1 and represses podocalyxin expression.

Wang D, Li Y, Wu C, Liu Y
PloS one 2011 Feb 24;6(2):e17048

Enhanced podocalyxin expression alters the structure of podocyte basal surface.

Economou CG, Kitsiou PV, Tzinia AK, Panagopoulou E, Marinos E, Kershaw DB, Kerjaschki D, Tsilibary EC
Journal of cell science 2004 Jul 1;117(Pt 15):3281-94

Overexpression of the anti-adhesin podocalyxin is an independent predictor of breast cancer progression.

Somasiri A, Nielsen JS, Makretsov N, McCoy ML, Prentice L, Gilks CB, Chia SK, Gelmon KA, Kershaw DB, Huntsman DG, McNagny KM, Roskelley CD
Cancer research 2004 Aug 1;64(15):5068-73

Human embryonal carcinoma tumor antigen, Gp200/GCTM-2, is podocalyxin.

Schopperle WM, Kershaw DB, DeWolf WC
Biochemical and biophysical research communications 2003 Jan 10;300(2):285-90

Human embryonal carcinoma tumor antigen, Gp200/GCTM-2, is podocalyxin.

Schopperle WM, Kershaw DB, DeWolf WC
Biochemical and biophysical research communications 2003 Jan 10;300(2):285-90

No comments: Submit comment

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of Human kidney lysates using Zymed Podocalyxin Monoclonal Antibody, Mouse (Product # 39-3800).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of Human kidney lysates using Zymed Podocalyxin Monoclonal Antibody, Mouse (Product # 39-3800).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis was performed on membrane enriched extracts (30 µg lysate) of NTERA-2 (Lane 1), HEK-293 (Lane 2), HeLa (Lane 3), Hep G2 (Lane 4), Jurkat (Lane 5) and PANC-1 (Lane 6). The blot was probed with Anti-Podocalyxin Mouse Monoclonal Antibody (Product # 39-3800, 2 µg/mL) and detected by chemiluminescence using Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, HRP conjµgate (Product # A28177, 0.4 µg/mL, 1:2500 dilution). A ~58 kDa band corresponding to Podocalyxin was observed across the cell lines tested. 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® 2 Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary Antibody using iBind™ Flex Western Starter Kit (Product # SLF2000S). 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 was performed using Anti-PODXL Monoclonal Antibody (3D3) (Product # 39-3800) and a 160 kDa band corresponding to PODXL was observed in all cell lysates tested except Hep G2 and A549. Whole cell lysates (30 µg lysate) of Hep G2 (Lane 1), NCCIT (Lane 2) and A549 (Lane 3) were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris gel (Product # NP0322BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (2 µg/mL) and detected by chemiluminescence with Goat anti-Mouse IgG (H+L), Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005). An uncharacterized band (*) was also observed at ~50 kDa.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis of Podocalyxin (PODXL) was performed using 90% confluent log phase HEK 293 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with PODXL (3D3) Mouse Monoclonal Antibody (Product # 39-3800) at 2µg/mL in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A28175) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Alexa Fluor® 555 Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing membrane localization. Panel e shows the no primary antibody control. The images were captured at 60X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis of PODXL was performed using 70% confluent log phase NCCIT and A549 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 1 hour at room temperature. The cells were labeled with PODXL Monoclonal Antibody (3D3) (Product # 39-3800) at 2 µg/mL in 0.1% BSA, incubated at 4 degree Celsius overnight and then labeled with Donkey anti-Mouse IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32766) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing localization to plasma membrane. Panel e shows A549 cells with no expression of PODXL. Panel f 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: Immunofluorescence analysis of Podocalyxin (PODXL) was performed in sections of iPSC derived kidney organoid. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes and blocked with 2% BSA for 1 hour at room temperature. The cells were labeled with PODXL (3D3) Mouse Monoclonal Antibody (Product # 39-3800) at 2 µg/mL in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A28175, 1:2,000 dilution) for 45 minutes at room temperature. The panels a & b represent the merged images PODXL monoclonal antibody 3D3 (green) (Product # 39-3800) with DAPI (blue) and F-actin (red) staining for iPSC control cells and differentiated kidney organoid. The images were captured at 20X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 7 PINCH1 blocks WT1-mediated podocalyxin expression in human podocytes. A , RT-PCR analyses demonstrate that PINCH1 blocked WT1-stimulated podocalyxin mRNA expression in podocytes. Cells were transfected with expression vectors for PINCH1, WT1 or both, respectively. RT-PCR amplification of housekeeping GAPDH was performed in an identical manner to serve as controls. B , Graphic presentation shows the relative PINCH1 mRNA abundance in different groups after normalization with GAPDH. Data are presented as mean +- SEM of three independent experiments. * P

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 1 Western blot analysis of podocalyxin (PODXL) expression in SW1783 and U-87 astrocytoma cells. (A) In SW1783 cells, PODXL expression was analyzed in normal control cells (NC), cells stably transfected with empty pcDNA3 vector (VC) and cells stably transfected with pcDNA3-PODXL expression vector (PODXL) with or without LY294002 (LY; 50 mu M) treatment by western blotting. In U-87 cells, PODXL expression was analyzed in NC, cells stably transduced with scramble control shRNA (SC) and cells stably transduced with PODXL-shRNA (P-shRNA) with or without LY (50 mu M) treatment by western blotting. beta-actin blotting was used as a loading control. Density of PODXL was normalized against that of beta-actin to obtain a relative blot density, which was expressed as fold change to the relative blot density of NC (designated as 1) in (B) SW1783 and (C) U-87 cells. a P

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Fig 1 Protein levels of podocalyxin (PODXL) and Bmi1 in oral tongue squamous cell carcinoma (OTSCC) cells with overexpression and knockdown of PODXL and/or Bmi1. In (A) SCC-4 and (B) Tca8113 OTSCC cells, the protein levels of PODXL and BMI1 were determined with western blot analysis in normal control cells (NC, lane 1), cells stably transfected with the empty pcDNA 3.1 vector (VC, lane 2), cells stably transfected with PODXL (lane 3), cells stably transfected with PODXL and treated with focal adhesion kinase (FAK) inhibitor 14 (50 M) for 24 hours (PODXL+FAK-I, lane 4), cells stably transfected with Bmi1 (lane 5), cells stably transduced with scramble control shRNA (SC, lane 6), cells stably transduced with PODXL-shRNA (lane 7), cells stably transduced with BMI1-shRNA (lane 8), cells stably transfected with PODXL and transduced with BMI1-shRNA (PODXL+BMI1-shRNA, lane 9), and cells stably transfected with Bmi1 and transduced with PODXL-shRNA (Bmi1+PODXL-shRNA, lane 10). Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) blotting was used as a loading control. Density of the Western blots was measured by densitometry, and the density of the PODXL and the Bmi1 blots was normalized against that of the GAPHD blot in the same sample to obtain a relative blot density to represent relative PODXL and Bmi1 content in each sample, respectively. a p

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 1 Immunohistochemical staining for podocalyxin (PODX) and Ki-67 in astrocytomas. Immunostaining for PODX (brown color) was observed in World Health Organization (WHO) grade (B) II, (E) III and (H) IV astrocytomas. Immunostaining for Ki-67 (brown color) was observed in WHO grade (C) II, (F) III and (I) IV astrocytomas. All the slides were counterstained with hematoxylin. The histotological images for WHO grades (A) II, (D) III and (G) IV astrocytomas are also shown. All images are x400 magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: 10.1371/journal.pone.0168346.g001 Fig 1 Urine cells on cytospin slides stained for podocalyxin (PCX, red), claudin-1 (CL1, green), and Dapi (blue). (A-D) A Faby podocyte which is PCX+/CL1- and shows characteristic vacuolated cytoplasm. (E-H) A urine cells which is PCX+/CL1+ consistent with a parietal epithelial cell (PEC) phenotype. (I-L) An apoptotic podocyte (PCX+/CL1-) in the urine from a Fabry patient with shrunken cytoplasm and small nucleus (arrow) compared to its adjacent cells. (M-P) A podocyte (PCX+/CL1-) in the urine from a healthy subject does not show vacuolated cytoplasm.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: 10.1371/journal.pone.0168346.g002 Fig 2 Urine cells on cytospin stained for podocalyxin (PCX, red), Dapi (blue), and TUNEL (green). Two podocytes (PCX+) with apoptotic nuclei (white arrows in B-D) are also positive for TUNEL (white arrows in C-D), confirming apoptosis. The yellow arrow (B-D) shows a non-podocyte cell (PCX-) that is TUNEL+ (C-D).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Fig. 2 Primary podocytes can be cultured from kidney organoid glomeruli. a Isolated organoid glomeruli show evidence of podocyte cell migration (OrgPods) displaying thin arborized projections (TAPs) (inset). Inverted image shown to provide maximum contrast, scale bar 100 um. b TAPS from newly emerged podocytes are composed of F-actin shown by phallodin immunofluorescent staining. Inverted image, scale bar 50 um. c Immunostaining at 36 h post-plating shows a strong positively stained 3D OrgGlom with a migrating 2D OrgPod population. Left panel 2D images, right panel 3D reconstruction of Z-stack. Scale bars 50 um. d At 48 h post-plating OrgPods display a flattened, arborized morphology with processes connecting adjacent cells (arrow), scale bar 50 um. e Immunostaining of ciPods for SYNAPTOPODIN showed expression is absent in undifferentiated cells (ciPod: Un), only becoming evident following 14 days induced differentiation at 37 degC (ciPod: Diff). OrgPods also display strong SYNAPTOPODIN protein expression, aligned with F-actin stress fibres. Scale bars 100 um. f OrgPods express the neonatal Fc receptor (FcRN) and actively endocytose fluorescein isothiocyanate (FITC)-labelled albumin at 37 degC resulting in FITC-accumulation in endosomes on the cell surface. This is process halted when performed at 4 degC. Scale bars 50 um. g OrgPods stimulated with insulin (10 mg/ml) for 10 min showed cortical reorganisation of their actin cytoskeleton with GLUT4 translocation from a vesicula

39-3800

Antibody data