Anti ck7

Manufactured by Abcam

Sourced in United Kingdom

Anti-CK7 is a primary antibody that recognizes the cytokeratin 7 (CK7) protein. CK7 is a type II cytokeratin expressed in a variety of epithelial cells, including those found in the lung, breast, ovary, and prostate. This antibody can be used for the detection of CK7 in immunohistochemical applications.

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4 protocols using anti ck7

Immunophenotyping of Stem-like Cancer Cells

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Immunophenotyping of 82.3 cells was performed by flow cytometric analysis. Cells were washed in 1× PBS containing 0.1% bovine serum albumin (BSA, Sigma-Aldrich, Saint Louis, MO, USA) and 0.01% sodium azide. For intracellular markers, cells were permeabilized with fix and perm reagent (BD Italia) following the manufacturer’s instructions. The following antibodies were used: FITC (Fluorescein isothiocyanate) conjugated mouse mAbs anti-CK7 and anti-CK19 (Abcam, Cambridge, UK) and anti-CD44 (BD Bioscience, San Jose, CA, USA), APC-(allophycocyanin) conjugated mouse mAbs anti-EPCAM and anti-AFP (BD Bioscience Europe), anti-CD34 and anti-CD133 (Miltenyi Biotec S.r.l., Bologna, Italy), PE (phycoerytrin) conjugated mAbs anti-CD24, anti-CXCR4, anti-Oct3/4, anti-FOXA1/2, anti-PDX1, (all from BD), anti-CD338 (R&D Systems, Inc., Minneapolis, MN, USA), PerCP-Cy 5.5 conjugated mAbs anti-SOX2/17, Alexa Fluor 647 conjugated mAbs anti-Nanog, anti-Stro1, and Alexa Fluor 488 conjugated mAbs anti-PAX6 (BD Bioscience Europe).

Peraldo-Neia C., Massa A., Vita F., Basiricò M., Raggi C., Bernabei P., Ostano P., Casorzo L., Panero M., Leone F., Cavalloni G, & Aglietta M. (2021). A Novel Multidrug-Resistant Cell Line from an Italian Intrahepatic Cholangiocarcinoma Patient. Cancers, 13(9), 2051.

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Immunohistochemistry Panel for Cancer Markers

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Immunohistochemistry was performed on standard tissue sections using an automated IHC stainer (Ventana, Tucson, AZ). In this study, we included 18 IHC markers. The primary antibodies used for the detection of these 18 proteins were as follows: anti-PAX-8 (Clone MRQ-50, Ventana), anti-P504S (Clone SP116, Ventana), anti-Ki-67 (Clone 30-9, Ventana), anti-CD10 (Clone SP67, Ventana), anti-HER2 (Clone 4B5, Ventana), anti-PTEN (Clone SP218, Ventana), anti-COX2 (Clone SP21, Ventana), anti-Vimentin (Clone Vim 3B4, Ventana), anti-TFE3 (Clone MRQ-37, Ventana), anti-CA9 (Cat No. 5649, Cell Signaling Technology), anti-CD117 (Cat No. 37805, Cell Signaling Technology), anti-mTOR (Cat No. 2983, Cell Signaling Technology), anti-CK7 (Cat No. ab181598, Abcam), anti-BAP1 (Cat No. ab199396, Abcam), anti-HGF (Cat No. ab83760, Abcam), anti-SETD2 (Cat No. PA5-43071, Invitrogen), anti-HIF-1α (Cat No. MA1-516, Invitrogen) and anti-PBRM1 (Cat No. HPA015629, Sigma-Aldrich). Criteria for the positive and negative scoring of IHC specimens for a given protein marker has been described previously (24 (link)), and all samples were evaluated by two independent experienced pathologists.

Wu J., Xu W.H., Wei Y., Qu Y.Y., Zhang H.L, & Ye D.W. (2018). An Integrated Score and Nomogram Combining Clinical and Immunohistochemistry Factors to Predict High ISUP Grade Clear Cell Renal Cell Carcinoma. Frontiers in Oncology, 8, 634.

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Western Blot for Liver Markers

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An equal amount of total protein was run on 10% SDS-PAGE (EpiZyme, cat. no. PG112), transferred to PVDF membranes (Merck millipore, cat. no. IPVH00010) (380 mA for 2 h), and probed with primary antibodies. The primary antibodies include anti-HBsAg (1:1000, 27 kDa, Novus Biologicals), anti-HBx (1:1000, 17 kDa, Abcam), anti-CK7 (1:1000, 51 kDa, Signalway antibody (SAB)), anti-CK19 (1:1000, 40 kDa, Signalway antibody (SAB)), anti-Hep-Par1 (1:1000, 165 kDa, Proteintech Group), anti-ALB (1:1000, 66 kDa, Proteintech Group), anti-α-Tubulin (1:1000, 55 kDa, Proteintech Group), anti-GAPDH (1:1000, 36 kDa, CST). The target protein bands were captured by binding of the secondary antibodies linked with peroxidase (1:1000, Anti-Rabbit IgG (H + L), CST) (1:1000, anti-mouse IgG (H + L), CST) to the primary antibodies.

Song Z., Lin S., Wu X., Ren X., Wu Y., Wen H., Qian B., Lin H., Huang Y., Zhao C., Wang N., Huang Y., Peng B., Li X., Peng H, & Shen S. (2023). Hepatitis B virus-related intrahepatic cholangiocarcinoma originates from hepatocytes. Hepatology International, 17(5), 1300-1317.

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Multiparametric Immunolabeling and Imaging Protocol

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For paraffin sections, dewaxing, hydration, and antigen retrieval steps were performed by following standard protocol. After blocking (10% normal goat serum [NGS]), slides were incubated with the primary antibodies as following: anti-pan-Kcr (PTM Biolabs 501 and 502, 1:1000 dilution); anti-Ehhadh (Santa Cruze sc-393123, 1:200 dilution); anti-CK7(Abcam ab181598, 1:2000 dilution); anti-Sox9 (Santa Cruze sc-166505, 1:100 dilution) overnight at 4 °C. The slides were then incubated with the secondary antibody (Goat anti-rabbit or -mouse IgG, Alexa Fluor 488 or 594; Thermo Fisher Scientific, 1:1000 dilution) for 2 h at 25 °C, followed by staining with DAPI (Thermo Fisher Scientific)/PBS for 6 min. Confocal imaging was performed using an SP8 system (Leica), and images were processed using the Leica AF software suite. For immunohistochemistry, anti-CD3 (99940, 1:150 dilution), anti-CD20 (70168, 1:500 dilution) and anti-CD68 (97778, 1:200 dilution) antibodies are from CST and anti-CD138 (Abcam ab128936, 1:500 dilution). Sections were incubated with biotinylated secondary antibodies and horseradish peroxidase avidin D (HRP, Vector). After three 5-min washes of PBS, the sections were developed with DAB kit (Vector) as standard procedures.

Zhang Y., Chen Y., Zhang Z., Tao X., Xu S., Zhang X., Zurashvili T., Lu Z., Bayascas J.R., Jin L., Zhao J, & Zhou X. (2022). Acox2 is a regulator of lysine crotonylation that mediates hepatic metabolic homeostasis in mice. Cell Death & Disease, 13(3), 279.

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