Cd4 4b12

Manufactured by Agilent Technologies

Sourced in United States

The CD4 (4B12) is a lab equipment product from Agilent Technologies. It is a reagent used for the identification and enumeration of T-helper/inducer lymphocytes in flow cytometric analysis.

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CD4 (clone 4B12, (11 citations)

5 protocols using cd4 4b12

Multiplex Immunohistochemistry for Tumor-Infiltrating Immune Cells

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One representative 4-μm-thick slide including the tumor and adjacent healthy pancreatic tissue was selected from each patient by two pathologists (M. L. and L. Y. C.). All of the slides were stained for CD4 (CD4/4B12, 1:100, Dako), CD8 (C8/144B, 1:100, Dako), CD68 (KP1, 1:100, Dako), and CD206 (ab64693, 1:2000, Abcam). Whole slides were prepared for immunohistochemistry according to the product protocol using monoclonal antibodies, as follows. After de-waxing, hydration, and antigen retrieval (EDTA at pH 8.0, 37 ℃ for 30 min), the immunohistochemistry protocol was performed using a Dako EnVision FLEX + detection system (DK-2600, Dako, Denmark) and an Ultraview Universal DAB Detection Kit (Ventana, AZ, US). All of the slides were re-stained with hematoxylin. Tonsil tissue was used as the positive control, and phosphate-buffered saline (instead of primary antibody) was used as the negative control.

Lu M., Zou Y., Fu P., Li Y., Wang P., Li G., Luo S., Chen Y., Guan G., Zhang S, & Chen L. (2023). The tumor–stroma ratio and the immune microenvironment improve the prognostic prediction of pancreatic ductal adenocarcinoma. Discover. Oncology, 14, 124.

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Immunohistochemical Analysis of Survivin

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Immunohistochemical detection of survivin was performed using a Dako Omnis autostainer (Dako North America, Inc. Carpinteria, CA) with rabbit monoclonal survivin antibody clone EP119 (Bio SB, Santa Barbara, CA). Additional antibodies included: CD4 (4B12, Dako); CD8 (C8/144B, Dako); CD20 (L26, Dako); PD-L1/CD274 (SP142, Spring Bioscience). Stained specimens were viewed by the neuropathologist, co-investigator (JQ), and survivin expression in the nucleus and cytoplasm was determined to be present or absent.

Fenstermaker R.A., Ciesielski M.J., Qiu J., Yang N., Frank C.L., Lee K.P., Mechtler L.R., Belal A., Ahluwalia M.S, & Hutson A.D. (2016). Clinical study of a survivin long peptide vaccine (SurVaxM) in patients with recurrent malignant glioma. Cancer Immunology, Immunotherapy, 65(11), 1339-1352.

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Immunohistochemical Analysis of Tumor Markers

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Immunohistochemistry was performed on 2‐μm FFPE sections in an automated immunostainer (Autostainer Link48; Dako, Glostrup, Denmark) according to the manufacturer's instructions. The following antibodies were used: β‐catenin 14 (1 : 1000; BD Transduction, South San Francisco, CA, USA), CD3 polyclonal (1 : 400; Dako), CD4 4B12 (1 : 300; Dako), CD8 C8/144B (1 : 20; Dako), CD20 L26 (1 : 400; Dako), CD21 1F8 (1 : 50; Dako), CD56 123C3 (1 : 400; Dako), CD68 KP1 (1 : 3000; Dako), CD163 10D6 (1 : 200; Novocastra, Newcastle, UK), FOXP3 259D/C7 (1 : 200; BD Pharmingen), PD‐L1 SP142 (1 : 30; SPRING, Pleasanton, CA, USA), PD1 NAT105 (1 : 100; Biocare Medical, Pikenine, Concord, NC, USA), and EZH2 5246S (1 : 50; Cell Signaling, Leiden, Netherlands).

Colombo C., Belfiore A., Paielli N., De Cecco L., Canevari S., Laurini E., Fermeglia M., Pricl S., Verderio P., Bottelli S., Fiore M., Stacchiotti S., Palassini E., Gronchi A., Pilotti S, & Perrone F. (2017). β‐Catenin in desmoid‐type fibromatosis: deep insights into the role of T41A and S45F mutations on protein structure and gene expression. Molecular Oncology, 11(11), 1495-1507.

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Histopathological and Ultrastructural Analysis of CAPNON

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The surgical resection or biopsy specimens were formalin fixed, routinely processed, paraffin-embedded, sectioned at 5 μm, and stained with hematoxylin and eosin (H&E) and by IHC. The H&E-stained slides were examined for histopathological features. Automated IHC was performed on tissue sections using the Dako Autostainer Link 48 and visualized with the Dako Envision Flex kit detection system (Dako, Carpinteria, CA, USA). Antibodies to the following were used: Neurofilament-light (NF-L; 2F11, Dako), Neurofilament-phosphorylated (NF-p; SMI 31, Covance, Berkeley, CA, USA), CD68 (KP1, Dako), CD8 (C8/144B, Dako), CD4 (4B12, Dako), glial fibrillary acidic protein (Z0334, Dako), Vimentin (V9, Dako), Desmin (D33, Dako), epithelial membrane antigen (E29, Dako), and synaptophysin (27G12, Leica). The antibodies listed here were only those immunostains described later in this study, while a few other immunostains and histological stains were also performed for the diagnosis of CAPNON in most cases. 3, 4 Electron Microscopy
In three selected cases of CAPNONs, a small portion of the tissue was fixed in 2% glutaraldehyde, postfixed in 1% osmium tetroxide, dehydrated, and embedded in resin. Ultrathin sections were stained with uranyl acetate followed by lead citrate and examined with JEOL 1230 Transmission electron microscope (EM).

Yang K., Reddy K., Wang B.H., Cenic A., Provias J., Popovic S., Yong W.H., Berthelet F., Bojanowski M.W., Hammond R, & Lu J.Q. (2021). Immunohistochemical Markers in the Diagnosis of Calcifying Pseudoneoplasm of the Neuraxis. The Canadian journal of neurological sciences. Le journal canadien des sciences neurologiques, 48(2).

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Comprehensive Immune Profiling of Melanoma

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Dual IHC for MHC class I (HLA-A, HLA-B, and HLA-C, clone EMR8-5, 1:6000; Abcam) or MHC class II (HLA-DP, HLA-DQ, HLA-DR, clone CR3/43, 1:750; Dako) with the melanoma marker SOX10 (EP268, 1:1500; Cell Marque) was performed using an automated staining system (Bond-III; Leica Biosystems), as previously described (42) .
IHC for CD3 (LN10; Leica), CD4 (4B12; Dako), CD8 (C8/144B; Dako), PD-1 (NAT105; Abcam), CD56 (clone 123C3; Dako), TCR (clone H-41; Santa Cruz Biotechnology), and 2M (A0072, 1:6000; Dako) was performed either manually (CD3, CD4, CD8, and PD-1) or on Bond RX (2M, CD56, and TCR) per standard protocols. IHC for PD-L1 (28-8; Dako) was performed as part of an investigational use-only kit (PD-L1 IHC pharmDx) on Dako Link 48 (2) .

Rodig S.J., Gusenleitner D., Jackson D.G., Gjini E., Giobbie-Hurder A., Jin C., Chang H., Lovitch S.B., Horak C., Weber J.S., Weirather J.L., Wolchok J.D., Postow M.A., Pavlick A.C., Chesney J, & Hodi F.S. (2018). MHC proteins confer differential sensitivity to CTLA-4 and PD-1 blockade in untreated metastatic melanoma. Science translational medicine, 10(450).

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