Pax8 10336 1 ap

Manufactured by Proteintech

Sourced in United States, Canada

The PAX8 (10336-1-AP) is a primary antibody produced by Proteintech. It is designed to detect the PAX8 protein, which is a transcription factor involved in the development and differentiation of various tissues, including the urogenital system, thyroid, and certain types of cancer. This antibody can be used in common laboratory techniques, such as western blotting, immunohistochemistry, and immunofluorescence, to identify and analyze the expression of the PAX8 protein.

Automatically generated - may contain errors

Lab products found in correlation

Vectashield h 1200, by Vector Laboratories (1 mentions) 6 well plate, by BD (1 mentions) Vimentin 5741, by Cell Signaling Technology (1 mentions) Goat serum, by Thermo Fisher Scientific (1 mentions) E cadherin ab15148, by Abcam (1 mentions) Vimentin v9 sc 6260, by Santa Cruz Biotechnology (1 mentions) Fluoroshield mounting medium with dapi, by Merck Group (1 mentions) Axin2 20540 1 ap, by Proteintech (1 mentions) Cell lytic m, by Merck Group (1 mentions) Trans blot system, by Bio-Rad (1 mentions) Novex 4 12 bis tris gel, by Thermo Fisher Scientific (1 mentions) β actin clone d6a8, by Cell Signaling Technology (1 mentions) Hsp90 clone c45g5, by Cell Signaling Technology (1 mentions) C myc clone y69, by Abcam (1 mentions) In cell analyzer 2000, by GE Healthcare (1 mentions) Pecam cd31, by BD (1 mentions) Donkey serum, by Merck Group (1 mentions) Myosin iib, by Cell Signaling Technology (1 mentions) B 1035, by Vector Laboratories (1 mentions) Ab15098, by Abcam (1 mentions)

6 protocols using pax8 10336 1 ap

Immunofluorescence Staining of Cultured Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

Cells were grown in 6-well plates (Falcon) coated with collagen IV and fixed with 4% PFA for 5 min, permeabilized with 0.3% Triton-X/PBS then blocked with 5% goat serum (Gibco, Life technologies) in PBS. Primary antibodies: C/EBPδ (SC-636), CK18 (M701029) (Dako, Agilent Technologies, Santa Clara, California, United States), Pax8 (10336-1-AP) (ProteinTech, Rosemont, IL, USA), E-cadherin (AB15148) (Abcam), Vimentin (5741S) (Cell Signaling), were applied overnight at 4 °C. Primary antibody was removed by washing samples with 1× PBS three times and was incubated with appropriate fluorophore-labeled secondary antibodies (Jackson ImmunoResearch Laboratories, West Grove, PA, USA) and Vectashield (H-1200, Vector Laboratories, Burlingame, CA, USA) in a dark area for 45 min. Cells were again washed for three times with 1× PBS and was mounted onto a coverslip and dried in the dark for 10 min. Samples were visualized using a Leica Axioimager (Leica).

Sowamber R., Chehade R., Bitar M., Dodds L.V., Milea A., Slomovitz B., Shaw P.A, & George S.H. (2019). CCAAT/enhancer binding protein delta (C/EBPδ) demonstrates a dichotomous role in tumour initiation and promotion of epithelial carcinoma. EBioMedicine, 44, 261-274.

+ Open protocol

+ Expand

Immunohistochemical and Western Blot Analyses

Check if the same lab product or an alternative is used in the 5 most similar protocols

WB and IHC analyses were performed using the following antibodies: p53 (D0-1, sc-126, Santa Cruz Biotechnology, Dallas, TX); HER2/ErbB2/Neu (C-18, sc-284, Santa Cruz Biotechnology, for WB; and OP15, Calbiochem®, Millipore (Canada) Ltd., Etobicoke, ON, for IHC); Cytokeratin (CK) 7 (Ab-2, MS-1352-P, Thermo Scientific, Waltham, MA); CK8 (Ab-4, MS-997-P, Thermo Scientific); CK18 (DC-10, sc-6259, Santa Cruz Biotechnology); CK19 (Ab-1, MS198-P, Thermo Scientific), WT1 (6F-H2, 05-753, Millipore (Canada) Ltd.), BRCA1 (OP92, MS110, Calbiochem®, Millipore (Canada) Ltd.), Vimentin (V9, sc-6260, Santa Cruz Biotechnology), E-cadherin (24E10, #3195, Cell Signaling Technology Inc., Danvers, MA), PAX8 (10336-1-AP, Proteintech, Chicago, IL) and beta-actin (AC-15, ab6276, Abcam Inc., Toronto, ON, Canada).

Fleury H., Communal L., Carmona E., Portelance L., Arcand S.L., Rahimi K., Tonin P.N., Provencher D, & Mes-Masson A.M. (2015). Novel high-grade serous epithelial ovarian cancer cell lines that reflect the molecular diversity of both the sporadic and hereditary disease. Genes & Cancer, 6(9-10), 378-398.

+ Open protocol

+ Expand

Immunohistochemical and Immunofluorescence Analysis of Stem Cell Markers in 3D Spheroids

Check if the same lab product or an alternative is used in the 5 most similar protocols

The expressions of PAX8, Axin2, ALDH1A1, and SOX9 were evaluated by IHC analysis. The FFPE spheroid sections (3 µm thick) were dewaxed in xylene and hydrated in graded alcohol. After antigen retrieval in sodium citrate buffer, the slides were incubated in overnight at 4°C with Abs at the following dilutions: 1:500 PAX8 (10336‐1‐AP; Proteintech), 1:200 Axin2 (20540‐1‐AP; Proteintech), 1:200 ALDH1A1 (15910‐1‐AP; Proteintech), and 1:500 SOX9 (HPA001758; Atlas Antibodies). Two gynecologic oncologists (S.S. and S.K.) independently evaluated the samples under a light microscope. Sections of normal endometria in the same patients were used as positive controls for all markers.
For immunofluorescence staining, FFPE spheroids or endometrium slides were loaded into a glass slide holder and dewaxed in xylene and then hydrated with alcohol. Sodium citrate buffer pH 6.0 was used for antigen retrieval in an autoclave for 10 min followed by incubation with primary Abs at a dilution of Axin2 1:200 and Ki‐67 1:100 overnight at 4°C followed by incubation with fluorescence‐labeled secondary Abs for 1 h at room temperature. Slides were then counterstained with Fluoroshield Mounting Medium with DAPI (Sigma‐Aldrich), and the immunofluorescence was detected using a Nikon Eclipse 50i fluorescence microscope with the appropriate filter.

Sato S., Nakayama K., Kanno K., Sultana R., Ishikawa M., Ishibashi T., Yamashita H, & Kyo S. (2023). Frequent PIK3CA mutation in normal endometrial gland drives spheroid formation and may be involved in stem cell propagation. Cancer Science, 114(6), 2335-2344.

+ Open protocol

+ Expand

Molecular Characterization of LVRCC67 Cell Line

Check if the same lab product or an alternative is used in the 5 most similar protocols

For validation of gene deletions and kidney cell origin of LVRCC67 cell line, whole cell extracts were prepared using CellLytic M (Sigma) and sonication for 10mins in a 4°c water bath. After centrifugation at 21,000rpm, supernatants were mixed with 6x loading dye containing 2-mercaptoethanol and run on pre-cast Novex 4–12% Bis-Tris gels (Invitrogen) for 90mins at 130V using the BioRad Transblot system. Proteins were transferred onto methanol-activated PVDF membranes for 1hour using 100V. Membranes were blocked with 5% milk and incubated at 4°c overnight in 5% BSA containing primary antibodies (Pax8 #10336-1-AP Proteintech 1:2500, HIF-1ɑ clone D1S7W Cell Signaling Technology 1:1000, c-myc clone Y69 Abcam 1:1000, Rb1 #9301S Cell signaling Technology 1:500, p53 clone 1C12 Cell Signaling Technology 1:1000, CD10 #PA5-47075 Invitrogen 1:2000, HSP-90 clone C45G5 Cell signaling Technology 1:3000, β-actin clone D6A8 1:5000) followed by secondary antibody incubation the following day for 1hour at room temperature.

Rappold P.M., Vuong L., Leibold J., Chakiryan N.H., Curry M., Kuo F., Sabio E., Jiang H., Nixon B.G., Liu M., Berglund A.E., Silagy A.W., Mascareno A., Golkaram M., Marker M., Reising A., Savchenko A., Millholland J., Chen Y.B., Russo P., Coleman J., Reznik E., Manley B.J., Ostrovnaya I., Makarov V., DiNatale R.G., Blum K.A., Ma X., Chowell D., Li M.O., Solit D.B., Lowe S.W., Chan T.A., Motzer R.J., Voss M.H, & Hakimi A.A. (2022). A Targetable Myeloid Inflammatory State Governs Disease Recurrence in Clear Cell Renal Cell Carcinoma. Cancer discovery, 12(10), 2308-2329.

+ Open protocol

+ Expand

Confocal Microscopy of Kidney Organoids

Check if the same lab product or an alternative is used in the 5 most similar protocols

For confocal microscopy, kidney organoids were differentiated on 96-well No, 1.5 coverslip glass-bottom plates (Mat-Tek). To fix, an equal volume of 8 % paraformaldehyde (Electron Microscopy Sciences) was added to the culture media (4 % final concentration) for 15 minutes at room temperature. After fixing, samples were washed in PBS, blocked in 5 % donkey serum (Millipore)/0.3 % Triton-X-100/PBS, incubated overnight in 3 % bovine serum albumin/PBS with primary antibodies, washed, incubated overnight with Alexa-Fluor secondary antibodies and DAPI (Invitrogen), and washed in PBS. Primary antibodies included ZO-1 (339100; Invitrogen), PAX8 (10336-1-AP, Proteintech), NPHS1 (AF4269, R&D), OAT1 (PA6-26244, Thermo Fisher), CLDN-1 (ab15098, Abcam), SYNPO (sc-21537; Santa Cruz), E-CAD (ab11512, Abcam), WT1 (sc-192; Santa Cruz), CFTR (570 antibody; University of North Carolina), Myosin IIB (3404S, Cell Signaling), CUBN (gift of Dr. Dennis Brown, Massachusetts General Hospital), AQP2 (HPA046834, Sigma), CD144/VE-cadherin (2500T, Cell Signaling), CD146/MCAM (ab75769, Abcam), and CD31/PECAM (555444; BD). LTL (FL-1321, Vector Laboratories) and DBA (B-1035, Vector Laboratories) were similarly applied. Fluorescence images were captured using an inverted Nikon epifluorescence Eclipse Ti or A1R confocal microscope. Automated imaging was performed using a GE INCELL 2000 Analyzer.

Czerniecki S.M., Cruz N.M., Harder J.L., Menon R., Annis J., Otto E.A., Gulieva R.E., Islas L.V., Kim Y.K., Tran L.M., Martins T.J., Pippin J.W., Fu H., Kretzler M., Shankland S.J., Himmelfarb J., Moon R.T., Paragas N.A, & Freedman B.S. (2018). High-throughput automation enhances kidney organoid differentiation from human pluripotent stem cells and enables multidimensional phenotypic screening. Cell stem cell, 22(6), 929-940.e4.

+ Open protocol

+ Expand

Immunohistochemical Analysis of Tumor Biopsy Samples

Check if the same lab product or an alternative is used in the 5 most similar protocols

Tumor biopsy samples were formalin fixed and paraffin embedded in the HUSLAB Pathology department according to standard procedures. For IHC analysis, formalin-fixed paraffin-embedded (FFPE) blocks were cut as 3.5μm sections and stained for antibodies against ALK (#3633, Cell Signaling Technology), CK7 (clone OV-TL 12/30, Agilent Dako), calretinin (clone DAK-Calret 1, Agilent Dako), WT1 (#83535, Cell Signaling Technology), CK5/6 (clone D5/16 B4, Agilent Dako), PAX8 (#10336-1-AP, ProteinTech,) and ER (clone 1D5, Agilent Dako) using protocols for routine diagnostics. IHC stained sections were scanned with a high-resolution whole-slide scanner (Pannoramic 250 Flash III, 3DHISTECH) with a x20 objective. The ALK rearrangement status of the tumor biopsy sample was evaluated by FISH in the HUSLAB Genetics laboratory using ALK dual-color break-apart probe according to manufacturer's protocol (Vysis, Abbott Molecular)

Murumägi A., Ungureanu D., Arjama M., Bützow R., Lohi J., Sariola H., Kanerva J., Koskenvuo M, & Kallioniemi O. (2021). STRN-ALK rearranged pediatric malignant peritoneal mesothelioma – Functional testing of 527 cancer drugs in patient-derived cancer cells. Translational Oncology, 14(4), 101027.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!