Ep1150y

Manufactured by Abcam

Sourced in United Kingdom

EP1150Y is a rabbit monoclonal antibody that recognizes the N-terminus of human PARP1 protein. It is intended for use in Western blot applications.

Automatically generated - may contain errors

Lab products found in correlation

Epr6855, by Abcam (3 mentions) Dm6000b, by Leica (1 mentions) Envision flex system, by Agilent Technologies (1 mentions) Meca 79, by BD (1 mentions) Epr1157 2, by Abcam (1 mentions) Nat105, by Abcam (1 mentions) High or low ph antigen retrieval buffer, by Agilent Technologies (1 mentions) Mib 1, by Agilent Technologies (1 mentions) Le cd19, by Agilent Technologies (1 mentions) Ab93278, by Abcam (1 mentions) Ab133616, by Abcam (1 mentions) Confocal microscope, by Leica (1 mentions) Ab134161, by Abcam (1 mentions)

5 protocols using ep1150y

Immunohistochemical Staining of Tumor Samples

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Tumors were recovered and processed from mice at necropsy. Samples were stained according to standard procedures. Briefly, resected tumors were fixed with 4% formalin, embedded in paraffin, and applied to produce sections by Biopathology Institute Co., Ltd (Servicebio, China). Primary antibodies used for IHC staining were polyclonal rabbit anti-human CD3 mAb (17617-1-AP, proteintech) or rabbit anti-human CD8 mAb (EP1150Y, Abcam). Images of IHC sections were obtained using the microscope (Leica, DM6000B).

Luo B., Zhan Y., Luo M., Dong H., Liu J., Lin Y., Zhang J., Wang G., Verhoeyen E., Zhang Y, & Zhang H. (2020). Engineering of α-PD-1 antibody-expressing long-lived plasma cells by CRISPR/Cas9-mediated targeted gene integration. Cell Death & Disease, 11(11), 973.

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Comprehensive Immunohistochemistry Profiling

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Tissue sections were deparaffinized in xylene and rehydrated through a graded ethanol series. Heat‐induced antigen retrieval was carried out in a high or low‐pH antigen retrieval buffer (DakoCytomation, Glostrup, Denmark). Endogenous peroxidase was blocked by incubating tissue sections in 3% H₂O₂ for 5 minutes. The primary antibodies against CD4 as a marker for helper T cells (1:500, EPR6855; Abcam, Cambridge, UK), CD8 as a maker for cytotoxic T lymphocytes (1:500, EP1150Y; Abcam), CD20 as a marker for pan B cells (1:50, L26; Abcam), Foxp3 as a marker for regulatory T cells (Treg) (1:300, 236A/E7; Abcam), PNAd as a marker for HEV (1:100, MECA‐79; BD Pharmingen^TM), programmed cell death (PD)‐1 as a marker for immune checkpoint molecules (1:50, NAT105; Abcam), Ki‐67 as a marker for cell proliferation (1:100, SP6; Abcam), CD80 as a marker for M1 macrophages (1:1000, EPR1157(2); Abcam) and CD163 as a marker for M2 macrophages (1:500, EPR11598; Abcam) were applied for 30 minutes. These sections were visualized using the HRP‐labeled polymer method (EnVision FLEX System, Dako). Immunostained sections were counterstained with hematoxylin, dehydrated in ethanol, and cleared in xylene. These data are summarized in Table S1.

Kuwabara S., Tsuchikawa T., Nakamura T., Hatanaka Y., Hatanaka K.C., Sasaki K., Ono M., Umemoto K., Suzuki T., Sato O., Hane Y., Nakanishi Y., Asano T., Ebihara Y., Kurashima Y., Noji T., Murakami S., Okamura K., Shichinohe T, & Hirano S. (2019). Prognostic relevance of tertiary lymphoid organs following neoadjuvant chemoradiotherapy in pancreatic ductal adenocarcinoma. Cancer Science, 110(6), 1853-1862.

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Multiparametric Analysis of Liver Biopsy

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Liver biopsy slides were shipped to Neogenomics (Fort Myers, FL). The MultiOmyx™ platform involves serial staining, imaging and dye-inactivation of two antigens at a time, one on Cy3 and another on Cy5 (Fig. S2).¹⁹ (link) The least robust targets are visualized first while the more robust targets are visualized last to limit the effects of dye-inactivation. Slides were stained in the following order: Round 1: anti-PD-L1 (73-10, Cell Signaling Technologies) and anti-CD299 (EPR11211, Abcam), Round 2: anti-PanCK (AE-1/AE-3, Biolegend) and anti-HBsAg (XTL17, Gilead), Round 3: anti-CD8 (EP1150Y, Abcam) and anti-HBcAg (366-2, Gilead), Round 4: anti-CD4 (EPR6855, Abcam) and anti-PD-1 (EH33, Cell Signaling Technologies), Round 5: anti-CD3 (97707, Abcam) and anti-FoxP3 (326A/E7, Abcam), Round 6: anti-CD19 (LE-CD19, Dako) and anti-Ki67 (MIB-1, Dako), Round 7: Anti-CD20 (L26, Dako) and anti-CD68 (KP-1, Diagnostic Biosystems). Images were collected on the GE InCell scanner at 20x. Quantitative morphometric image analysis was performed both at Neogenomics using a proprietary software platform, and at Gilead using Visiopharm.

van Buuren N., Ramirez R., Turner S., Chen D., Suri V., Aggarwal A., Moon C., Kim S., Kornyeyev D., Bui N., Bhardwaj N., Chan H.L., Marcellin P., Buti M., Wallin J., Gaggar A., Fletcher S.P., Diehl L., Li L., Mo H, & Feierbach B. (2021). Characterization of the liver immune microenvironment in liver biopsies from patients with chronic HBV infection. JHEP Reports, 4(1), 100388.

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Quantifying Tumor Infiltrating Lymphocytes

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Immunohistochemistry (IHC) analysis on the archived tumor samples was applied to compare the expression level of CD3, CD4, and CD8 in tumor infiltrated lymphocytes. Anti-CD8 (EP1150Y, ab93278), anti-CD4 (EPR6855, ab133616), and anti-CD3 (SP7, ab16669) antibody were purchased from Abcam (Cambridge, MA). All experiments were undergone following the instructor’s protocol. Two pathologists independently interpreted IHC staining by assessing background staining, positive and negative controls, and localization and amount of biomarker staining in all specimens. Percent positive cells = (number of positive lymphocytes/tumor area occupied by tumor cells, associated intratumoral, and contiguous peritumoral stroma) × 100.

Niu S., Liu K., Xu Y., Peng C., Yu Y., Huang Q., Wu S., Cui B., Huang Y., Ma X., Zhang X, & Wang B. (2021). Genomic Landscape of Chinese Clear Cell Renal Cell Carcinoma Patients With Venous Tumor Thrombus Identifies Chromosome 9 and 14 Deletions and Related Immunosuppressive Microenvironment. Frontiers in Oncology, 11, 646338.

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Immunohistochemical Staining of Tumor Tissue

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IHC staining was performed following a previously established protocol [42 (link)]. Tumor tissues were fixed in 10% buffered formalin overnight, dehydrated in an ethanol series, cleared in xylene, and embedded in paraffin wax. Sections were cut at 5-μm thickness and stained with CD8 (Abcam, EP1150Y), PD-L1(NOVUS, 13684T), and B7-H3 monoclonal antibodies (Abcam, ab134161). The sections were mounted, viewed, and photographed with a confocal microscope (Leica, Germany).

Shao L., He Q., Wang J., He F., Lin S., Wu L., Gao Y., Ma W., Dong J., Yang X, & Li F. (2021). MicroRNA-326 attenuates immune escape and prevents metastasis in lung adenocarcinoma by targeting PD-L1 and B7-H3. Cell Death Discovery, 7, 145.

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