- PA1-108 - Invitrogen Antibodies PAX4 antibody

Zhang Y, Ding L, Ni Q, Tao R, Qin J
Aging 2021 Jun 23;13(12):16786-16803

Sandilya S, Singh S
Cell regeneration (London, England) 2021 Dec 1;10(1):38

Tanaka T, Kojima D, Mera T, Matsumoto M, Yasunami Y, Yanase T
Heliyon 2018 May;4(5):e00632

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of Pax4 was performed by loading the indicated amounts of recombinant Pax3 (negative control) or Pax4 proteins, and 10 µL of PageRuler Prestained Protein Ladder (Product # 26616) per well onto a Novex® 4-20% Tris-Glycine polyacrylamide gel. Proteins were transferred to a PVDF membrane using the G2 Fast Blotter (Product # 62288), and blocked with 5% milk in TBST for at least 1 hour at room temperature. Pax4 was detected at ~38 kD using a Pax4 polyclonal antibody (Product # PA1-108) at a dilution of 1:1000 in blocking buffer overnight at 4C on a rocking platform, followed by an HRP-conjugate goat anti-rabbit IgG secondary antibody (Product # 31460) at a dilution of 1:40,000 for at least 30 minutes. Chemiluminescent detection was performed using SuperSignal West Pico (Product # 34080). NOTE: The presence of Pax3 recombinant protein was determined by probing a duplicate blot with a Pax3 polyclonal antibody (Product # PA1-107), and the corresponding image is on the webpage for Product # PA1-107.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of Pax4 was performed by loading the indicated amounts of recombinant Pax3 (negative control) or Pax4 proteins, and 10 µL of PageRuler Prestained Protein Ladder (Product # 26616) per well onto a Novex® 4-20% Tris-Glycine polyacrylamide gel. Proteins were transferred to a PVDF membrane using the G2 Fast Blotter (Product # 62288), and blocked with 5% milk in TBST for at least 1 hour at room temperature. Pax4 was detected at ~38 kD using a Pax4 polyclonal antibody (Product # PA1-108) at a dilution of 1:1000 in blocking buffer overnight at 4C on a rocking platform, followed by an HRP-conjugate goat anti-rabbit IgG secondary antibody (Product # 31460) at a dilution of 1:40,000 for at least 30 minutes. Chemiluminescent detection was performed using SuperSignal West Pico (Product # 34080). NOTE: The presence of Pax3 recombinant protein was determined by probing a duplicate blot with a Pax3 polyclonal antibody (Product # PA1-107), and the corresponding image is on the webpage for Product # PA1-107.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence was performed on adult human pancreas tissue. To expose target proteins, antigen retrieval was performed by heating tissues at 95C for 15 minutes in 10mM citrate buffer, pH 6.0. Following antigen retrieval, tissues were blocked and then probed with a Pax4 polyclonal antibody (Product # PA1-108) at a dilution of 1:500 overnight at 4C. Tissues were washed extensively and detection was performed using a fluorescently-conjugated anti-rabbit IgG secondary antibody (green). Tissues were also stained with an antibody against insulin (red), and nuclei (grey) were stained with DAPI. Higher magnification of regions inside the white boxes (left panel) are shown to the right. Scale bar = 200 µm. Data supplied courtesy of the Innovators Program.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence was performed on adult mouse pancreas tissue. To expose target proteins, antigen retrieval was performed by heating tissues at 95C for 15 minutes in 10mM citrate buffer, pH 6.0. Following antigen retrieval, tissues were blocked and then probed with a Pax4 polyclonal antibody (Product # PA1-108) at a dilution of 1:500 overnight at 4C. Tissues were washed extensively and detection was performed using a fluorescently-conjugated anti-rabbit IgG secondary antibody (green). Tissues were also stained with an antibody against insulin (red), and nuclei (grey) were stained with DAPI. Higher magnification of regions inside the white boxes (left panel) are shown to the right. Scale bar = 200 µm. Data supplied courtesy of the Innovators Program.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 1 Elevated PAX4 mRNA and protein expression in both GC tissues and cell lines. ( A , B ) Higher PAX4 mRNA and protein expression in GC tissues compared to normal tissues ( n = 60) by qRT-PCR ( P < 0.0001) and WB ( P = 0.0002). ( C ) The IHC results between GC and non-GC tissues demonstrated excessive PAX4 in GC tissues. ( D ) Overexpression of PAX4 mRNA levels were identified across six GC cell lines (HGC-27, MGC803, BGC-823, SGC-7901, AGS, MKN45) compared to the human gastric mucosal epithelial cell line GES-1. ( E ) The overall survival analysis results demonstrated that PAX4 high expression in GC is associated with poor prognosis.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Fig. 2 Stage wise Immunohistochemistry of the hiPSC cells undergoing differentiation. Foregut lineage cells show positive staining for PDX1 and faint staining for few progenitor markers. Pancreatic progenitor cells show positive staining for all the stage specific markers

PA1-108