Ab93650

Manufactured by Abcam

Sourced in United Kingdom

Ab93650 is a polyclonal antibody that targets the human PCNA (Proliferating Cell Nuclear Antigen) protein. It is intended for use in various research applications, such as western blotting, immunohistochemistry, and immunocytochemistry, to detect and study the PCNA protein.

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4 protocols using ab93650

Immunohistochemical Analysis of Kidney Samples

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PAS and H&E staining were performed according to standard procedures. Paraffin sections were placed in citrate-buffered solution (pH 6.0) and boiled for 10 min for antigen retrieval. Endogenous peroxidase was blocked with 3% hydrogen peroxide. Samples were blocked with 3% BSA in PBS and incubated with primary antibodies, namely Rabbit anti–Podocin (1:500; ab93650, Abcam, Cambridge, MA), Aquaporin 1 (1:500; ab9566, Abcam), CD68 (1:500; ab31630, Abcam), PHH3 (1:500; #9706s, Cell Signaling Technology, Danvers, MA), and the lectin, Dolichos biflorus Agglutinin (DBA, 1:500; Vector Labs, Burlingame, CA). Diaminobenzidine substrate (Dako, Hamburg, Germany) was used for the color reaction. Sections were counterstained with hematoxylin. Secondary antibody alone was consistently negative on all sections.

Kirita Y., Kami D., Ishida R., Adachi T., Tamagaki K., Matoba S., Kusaba T, & Gojo S. (2016). Preserved Nephrogenesis Following Partial Nephrectomy in Early Neonates. Scientific Reports, 6, 26792.

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Podocyte Protein Expression Analysis

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Total protein was isolated from podocytes with RIPA (Beyotime, Shanghai, China) and quantified using a BCA Protein Quantification Kit (Tiangen, Beijing, China). Then 30-μg proteins were electrophoresed through 10% SDS-PAGE (Solarbio, Beijing, China) and blotted on polyvinylidene fluoride. After being blocked in 5% slim milk for 1 h, the membranes were blotted with primary antibodies against alpha-smooth muscle actin (α-SMA; ab32575; Abcam, Cambridge, Massachusetts, United States), fibronectin (FN; ab2413; Abcam), collagen I (Col I; ab34710; Abcam), podocin (ab93650; Abcam), TIMP3 (ab39814; Abcam) and β-actin (ab8228; Abcam) overnight at 4°C followed by interaction with horseradish peroxidase-conjugated secondary antibody (ab205719; Abcam) for 1.5 h at indoor temperature. The proteins were developed with an ECL reagent (Vazyme).

Fan H, & Zhang W. (2020). Overexpression of Linc 4930556M19Rik Suppresses High Glucose-Triggered Podocyte Apoptosis, Fibrosis and Inflammation via the miR-27a-3p/Metalloproteinase 3 (TIMP3) Axis in Diabetic Nephropathy. Medical Science Monitor : International Medical Journal of Experimental and Clinical Research, 26, e925361-1-e925361-11.

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Losartan and Puerarin in Diabetic Nephropathy

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Losartan was purchased from Merck & Co Inc (H20000371). Puerarin was purchased from NANJING CHIA TAI TIANQING (06060521). All other chemicals and reagents were purchased from Sigma-Aldrich, Shanghai, China. Antibodies against GAPDH (ab37168), nephrin (rabbit, ab58968), podocin (rabbit, ab93650), and MMP-9 (rabbit, ab7299) were purchased from Abcam, Cambridge, UK. Anti-S-Nitroso-Cysteine antibody is obtained from Sigma-Aldrich. Antibody for type IV collagen was obtained from Chemicon. Horseradish Peroxidase (HRP) conjugated goat anti-rabbit secondary antibody (ab6721) was also purchased from the Abcam.

Zhong Y., Zhang X., Cai X., Wang K., Chen Y, & Deng Y. (2014). Puerarin Attenuated Early Diabetic Kidney Injury through Down-Regulation of Matrix Metalloproteinase 9 in Streptozotocin-Induced Diabetic Rats. PLoS ONE, 9(1), e85690.

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Immunohistochemical Analysis of Kidney Markers

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Immunohistochemical staining was performed using a routine protocol. Formalin-fixed, paraffin-embedded sections (3 μm) were deparaffinized and rehydrated. Following antibodies were applied overnight at 4°C, including anti-fibronectin (ab23751, Abcam, Cambridge, UK), anti-collagen IV (ab6586, Abcam, Cambridge, UK), anti-podocin (ab93650, Abcam, Cambridge, UK), and anti-nephrin (ab183099, Abcam, Cambridge, UK). Anti-rabbit (goat polyclonal) antibodies were applied subsequently for 30 min. Slides were visualized using an Olympus light microscope, and quantification of intensity for each condition was performed using ImageJ software.

Wei M.G., He W.M., Lu X., Ni L., Yang Y.Y., Chen L., Xiong P.H, & Sun W. (2016). JiaWeiDangGui Decoction Ameliorates Proteinuria and Kidney Injury in Adriamycin-Induced Rat by Blockade of TGF-β/Smad Signaling. Evidence-based Complementary and Alternative Medicine : eCAM, 2016, 5031890.

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