Ab46799

Manufactured by Abcam

Sourced in United Kingdom

Ab46799 is a primary antibody that can be used in immunoassays. It is a monoclonal antibody produced in mouse and purified from mouse ascites fluid.

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Lab products found in correlation

8 protocols using ab46799

Antibody Panel for Liver Cancer Markers

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Antibodies for CD3 (Ab5690), CD4 (Ab183685), CD68 (Ab125212), alpha fetoprotein (α-AFP) (Ab46799), and Glypican-3 (GPC-3) (Ab66596) were purchased from Abcam Inc. (Cambridge, MA). Antibodies for SV40 TAg (v-300) (sc-20800) were purchased from Santa Cruz Biotechnology, Inc. (Dallas, TX). Antibodies for CD8a (14-0808) and F4/80 (14-4801-81) were purchased from Affymetric eBioscience, (San Diego, CA).

Qi X., Lam S.S., Liu D., Kim D.Y., Ma L., Alleruzzo L., Chen W., Hode T., Henry C.J., Kaifi J., Kimchi E.T., Li G, & Staveley-O’Carroll K.F. (2016). Development of inCVAX, In situ Cancer Vaccine, and Its Immune Response in Mice with Hepatocellular Cancer. Journal of clinical & cellular immunology, 7(4), 438.

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Antibody Panel for Liver Cancer Markers

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Antibodies for Cell Signaling Analysis

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Antibodies against Cnot1, Cnot2, Cnot6L, Cnot7, Cnot8 and Cnot9 have been described previously (Suzuki et al., 2015 (link)). Antibodies against α-tubulin (DM1A, sc-32293), Cdt1 (sc-365305), Brca1 (sc-135731), Alb (sc-46291, for WB), normal mouse IgG (sc-2025) and normal rabbit IgG (sc-2026) were purchased from Santa Cruz Biotechnology. Antibodies against cleaved caspase-3 (9664) and p53 (2524) were from Cell Signaling Technology. Anti-Ki67 rabbit monoclonal antibody for immunohistochemistry was from NeoMarkers (RM-9106-S0). Antibodies against CK19 (ab133496), Ki67 (ab16667), phospho-histone H3 (Ser10) (ab5176), Afp (ab46799), Igf2 (ab9574), CD45 (ab10558) and F4/80 (ab111101) were from Abcam. Anti-Alb antibody (MAB-1455, for immunofluorescence) was from B&D systems. Antibodies against Chk1 (K0086-3) and Iqgap1 (K0100-3) were from MBL. Anti-Klf6 antibody (14716-1-AP) was from Proteintech.

Suzuki T., Kikuguchi C., Nishijima S., Nagashima T., Takahashi A., Okada M, & Yamamoto T. (2019). Postnatal liver functional maturation requires Cnot complex-mediated decay of mRNAs encoding cell cycle and immature liver genes. Development (Cambridge, England), 146(4), dev168146.

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Immunohistochemical Analysis of Liver Tissues

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Mice were sacrificed at 5 weeks, 3 months, or 5 months post-injection. Livers were removed and fixed overnight in 4% formalin, then embedded in paraffin, and cut into 5-μm sections. Immunohistochemistry was performed using the EnVision FLEX Mini Kit (DAKO). Antigen retrieval was done in a PT Link machine (DAKO). The primary antibodies used for immunohistochemistry are rabbit polyclonal anti-FAH (ThermoFisher Scientific, PA5-42049, 1:100), incubated for 120 min, and rabbit polyclonal anti-alpha 1 fetoprotein (Abcam, ab46799, 1:500) and rabbit polyclonal anti-glypican 3 (Abcam, ab186872, 1:800), incubated overnight. Secondary antibody polyclonal goat anti-rabbit-HRP (DAKO, P0448) was incubated for 30 min. Visualization was done with EnVision FLEX DAB+ Chromogen System (DAKO, GV825). After hematoxylin counterstaining for 5 min, slides were mounted and scanned with a Pannoramic Digital Slide Scanner (3D Histech).

Kopasz A.G., Pusztai D.Z., Karkas R., Hudoba L., Abdullah K.S., Imre G., Pankotai-Bodó G., Migh E., Nagy A., Kriston A., Germán P., Drubi A.B., Molnár A., Fekete I., Dani V.É., Ocsovszki I., Puskás L.G., Horváth P., Sükösd F, & Mátés L. (2022). A versatile transposon-based technology to generate loss- and gain-of-function phenotypes in the mouse liver. BMC Biology, 20, 74.

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Immunohistochemical Analysis of Tissue Samples

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Tissue samples were fixed in 4% paraformaldehyde (PFA, Alfa Aesar, Ward Hill, MA) overnight at 4 °C. Tissues were then dehydrated in 70% ethanol until processing in paraffin. Samples were processed in the Texas Medical Center Digestive Diseases Center (Houston, TX). H&E and immunohistochemical staining of AFP (CP028, Biocare Medical, Concord, CA (Supplementary Fig. S1a); AC-0166, Epitomics, Cell Marque, Rocklin, CA (Supplementary Fig. S1c); ab46799, Abcam, Cambridge, MA (Supplementary Fig. S1b); 180003, Thermo Fisher Scientific, Waltham, MA (Fig. 4e)), β-catenin (ab32573, Abcam), and GPC3 (GPC3, 11395, Santa Cruz Biotechnology, Dallas, TX) were performed. Imaging of tumor sections on slides was done on a DMi8 microscope (Leica, Germany).

Woodfield S.E., Shi Y., Patel R.H., Jin J., Major A., Sarabia S.F., Starosolski Z., Zorman B., Gupta S.S., Chen Z., Ibarra A.M., Bissig K.D., Ghaghada K.B., Sumazin P., López-Terrada D, & Vasudevan S.A. (2017). A Novel Cell Line Based Orthotopic Xenograft Mouse Model That Recapitulates Human Hepatoblastoma. Scientific Reports, 7, 17751.

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Immunohistochemical Analysis of HCC Tissues

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Immunohistochemistry staining of IL-17 and AFP was performed on surgically resected HCC tissues preserved in the formalin-fixed paraffin-embedded (FFPE) blocks and cut into sections with a thickness of 5 μm. The FFPE samples were deparaffinized, dehydrated and incubated with the appropriate antibodies (ab136668 and ab46799, Abcam, Cambridge, UK) for the staining.

Liang K.H., Lai M.W., Lin Y.H., Chu Y.D., Lin C.L., Lin W.R., Huang Y.H., Wang T.H., Chien R.N., Hu T.H, & Yeh C.T. (2021). Plasma interleukin-17 and alpha-fetoprotein combination effectively predicts imminent hepatocellular carcinoma occurrence in liver cirrhotic patients. BMC Gastroenterology, 21, 177.

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Quantifying Liver Tumor Characteristics

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At the end of the 40-week study, the remaining whole livers were fixed (10% NBF), and tumors were microdissected out of the liver, counted, and sized. Tumor volume was calculated using the modified ellipsoid method [0.5 (length × width²)] after sizing with a digital caliper. Tumor markers alpha-fetoprotein (ab46799, Abcam, 1:100 dilution) and p62 (23214, Cell Signaling Technology, 1:250 dilution) were detected within dewaxed, rehydrated liver FFPE sections and stained overnight at 4°C in blocking reagent (Ultracruz, Santa Cruz Biotechnology) following heat-based antigen retrieval (15 min, 95°C in 10 mM sodium citrate buffer with buffer refresh at 10 min). Tumor markers were visualized with chromogen DAB/0.5% hydrogen peroxide [0.1 M tris-HCl (pH 7.6)] following horseradish peroxidase (HRP)–conjugated secondary antibody incubation (90 min, GTX213110-01, GeneTex, 1:250) and three PBS washes. Sections were counterstained with hematoxylin, washed in distilled water, dehydrated through ethanol and xylene, and then mounted in limonene organic mounting media (Santa Cruz Biotechnology) before visualization via light microscopy. Area of brown DAB staining was quantified (FIJI software).

Boslem E., Reibe S., Carlessi R., Smeuninx B., Tegegne S., Egan C.L., McLennan E., Terry L.V., Nobis M., Mu A., Nowell C., Horadagoda N., Mellett N.A., Timpson P., Jones M., Denisenko E., Forrest A.R., Tirnitz-Parker J.E., Meikle P.J., Rose-John S., Karin M, & Febbraio M.A. (2023). Therapeutic blockade of ER stress and inflammation prevents NASH and progression to HCC. Science Advances, 9(37), eadh0831.

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Histological Assessment of Liver Carcinoma

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Sections of formalin-fixed, paraffin-embedded livers were stained with hematoxylin and eosin (H&E), or Sirius Red and assessed for histological features of carcinoma and fibrosis. The H&E stained sections were independently examined by a veterinary pathologist, Dr. Garlick in a blinded manner (see acknowledgments). The quantitation of Sirius Red staining was performed using ImageJ software. Immunohistochemistry staining for AFP (ab46799; Abcam), CK7 (ab9021; Abcam), CK19 (ab52625; Abcam), was performed on formalin-fixed, paraffin-embedded livers according to the manufacturer’s instructions. To examine cell proliferation, mice were injected i.p. with 100 mg/kg BrdU (Sigma, St. Louise, MO) 2 hr prior to sacrifice, and paraffin sections were stained using the anti BrdU antibody (ab6326, Abcam). ImageJ (NIH) was used for image analysis.

Ambade A., Satishchandran A, & Szabo G. (2016). Alcoholic hepatitis accelerates early hepatobiliary cancer by increasing stemness and miR-122-mediated HIF-1α activation. Scientific Reports, 6, 21340.

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