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Cd4 clone 4b12

Manufactured by Agilent Technologies
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Sourced in United Kingdom, United States, Denmark

The CD4 (clone 4B12) is a laboratory reagent used for the identification and enumeration of CD4-positive cells in biological samples. It is designed for use in flow cytometry applications.

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Lab products found in correlation

Cd8 clone c8 144b, by Agilent Technologies (7 mentions) Cd20 clone l26, by Agilent Technologies (6 mentions) Granzyme b clone grb 7, by Agilent Technologies (3 mentions) Autostainer link 48, by Agilent Technologies (3 mentions) Axiocam mrc camera, by Zeiss (2 mentions) Dako autostainer link, by Agilent Technologies (2 mentions) Cd19 clone le cd19, by Agilent Technologies (2 mentions) Cd38 clone spc 32, by Leica (2 mentions) Bx51 microscope, by Olympus (2 mentions) Cd3 polyclonal, by Agilent Technologies (2 mentions) Cd16 clone 2h7, by Leica (2 mentions) Primary monoclonal antibodies, by Agilent Technologies (1 mentions) Automated immunostainer, by Agilent Technologies (1 mentions) Cd3 clone f7.2.38, by Agilent Technologies (1 mentions) Cd30 clone berh2, by Agilent Technologies (1 mentions) Envision flex target retrieval solution, by Agilent Technologies (1 mentions) Envision detection kit, by Agilent Technologies (1 mentions) Axio scan z1 digital slide scanner, by Zeiss (1 mentions) Pd 1 clone nat105, by Abcam (1 mentions) Cd8 clone 1a5, by BioGenex (1 mentions)

Spelling variants of this product

Clone 4B12 (6 citations) CD4 (4B12, (5 citations)

11 protocols using cd4 clone 4b12

1

Immunohistochemical Analysis of Tissue Samples

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Bone marrow biopsies were decalcified in EDTA prior to paraffin embedding. 3 μm sections were cut from formalin-fixed, paraffin-embedded samples. Upper gastrointestinal biopsies were routinely stained with hematoxylin and eosin stain and periodic acid–Schiff stain. Bone marrow biopsies were routinely stained with naphthol AS-D chloroacetate esterase (NASDCL). Immunohistochemical staining was performed using a horseradish peroxidase catalyzed brown chromogen reaction together with ready-to-use antibodies in an automated staining system (Dako Autostainer Link®; Dako, Glostrup, Denmark), following the manufacturer's guidelines. Depending on the tissue section size, up to three droplet zones are stained with 100 μl antibody solution per droplet zone. The following antibodies were used: CD3 (rabbit polyclonal, Dako), CD4 (clone 4B12, Dako), CD8 (clone C8/144B, Dako), CD19 (clone LE-CD19, Dako), CD20 (clone L26, Dako), CD38 (clone SPC 32, Novocastra, Newcastle upon Tyne, UK), IgM, IgG, IgA (rabbit polyclonal antibodies; Dako). Photos were taken on an Olympus BX51 microscope (Olympus Germany, Hamburg, Germany) with the AxioCam MRc camera (Zeiss, Jena, Germany).
Schubert D., Bode C., Kenefeck R., Hou T.Z., Wing J.B., Kennedy A., Bulashevska A., Petersen B.S., Schäffer A.A., Grüning B.A., Unger S., Frede N., Baumann U., Witte T., Schmidt R.E., Dueckers G., Niehues T., Seneviratne S., Kanariou M., Speckmann C., Ehl S., Rensing-Ehl A., Warnatz K., Rakhmanov M., Thimme R., Hasselblatt P., Emmerich F., Cathomen T., Backofen R., Fisch P., Seidl M., May A., Schmitt-Graeff A., Ikemizu S., Salzer U., Franke A., Sakaguchi S., Walker L.S., Sansom D.M, & Grimbacher B. (2014). Autosomal-dominant immune dysregulation syndrome in humans with CTLA4 mutations. Nature medicine, 20(12), 1410-1416.
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2

Immunohistochemical Analysis of Metastatic Melanoma

Cited 1 time
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NV mice were sacrificed on days 0, 7 and 14 after B16OVA melanoma transplantation. The most common metastatic organs (liver, spleen, kidney, adrenal glands, liver and lungs) were resected, sectioned, and fixed by immersion in 4% formaldehyde for 24 h. Organs were subsequently embedded in paraffin, processed, and sections stained with hematoxylin–eosin) and immunohistochemical analysis of lymphocyte markers performed as previously described [10 (link)]. Primary monoclonal antibodies (Dako, Carpinteria, CA, USA) used were against the following antigens, CD4 (clone 4B12), CD8 (clone c8/144B), CD23 (clone DAK-CD23), CD45 (clone 2B11 + PD7/26), CD56 (clone 123C3) and CD68 (clone KP1) and visualized as described [10 (link)].
Calderon-Gonzalez R., Bronchalo-Vicente L., Freire J., Frande-Cabanes E., Alaez-Alvarez L., Gomez-Roman J., Yañez-Diaz S, & Alvarez-Dominguez C. (2016). Exceptional antineoplastic activity of a dendritic-cell-targeted vaccine loaded with a Listeria peptide proposed against metastatic melanoma. Oncotarget, 7(13), 16855-16865.
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3

Comprehensive Immunohistochemistry Profiling

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Immunohistochemistry on cell block and FFPE capsular specimens was performed using an automated immunostainer (Dako, Glostrup, Denmark) with the following primary antibodies: CD30 (clone Ber-H2), CD3 (clone F7.2.38), CD4 (clone 4B12), CD8 (clone C8/144B), CD68 (clone PG1), CD15 (clone Car-b), Granzyme B (clone GrB-7), IRF-4 (clone MUM1) (Dako), CD25 (clone 4C9, Novocastra, Newcastle Upon Tyne, UK), PAX5 (clone SP34, Thermo Scientific, Waltham, USA). Paraffin sections were pretreated using EnVision™ FLEX Target Retrieval Solution (Dako) and incubated with an optimal dilution of the primary antibody. The reaction was visualized with the EnVision Detection Kit (Dako) using 3–3’-diaminobenzidine chromogenic substrate. Sections were counterstained with EnVision FLEX Hematoxylin (Link) (Dako).
For every marker, positive cells were counted out of 10 non-overlapping randomly selected high-power microscopic fields (HPFs, 40×10), and the mean number of positively stained cells per HPF was recorded. The percentage of positive cells was calculated as the ratio between the mean number of stained cells per HPF and the mean number of total cells per HPF.
Di Napoli A., Pepe G., Giarnieri E., Cippitelli C., Bonifacino A., Mattei M., Martelli M., Falasca C., Cox M.C., Santino I, & Giovagnoli M.R. (2017). Cytological diagnostic features of late breast implant seromas: From reactive to anaplastic large cell lymphoma. PLoS ONE, 12(7), e0181097.
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4

Quantifying Immune Cell Profiles in Primary Tumors

Cited 1 time
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Immunohistochemistry (IHC) was performed on formalin-fixed, paraffin-embedded primary tumor tissue using the antibodies: CD3 (clone IR503; DAKO), CD8a (clone: C8/144B; DAKO), CD4 (clone 4B12; DAKO), CD103 (clone EPR4166(2); Abcam), PD-1 (clone NAT105; Abcam), PD-L1 (clone E1L3N; CST), MHCI ABC (clone EMR8-5; Abcam) and MHCII DR+DP+ DQ (clone: CR3/43; Abcam). Stained slides were scanned using a Zeiss Axio Scan.Z1 Digital Slide Scanner and Zeiss ZEN software (V.2.6) and positive cells were enumerated using the QuPath bioimage analysis software (V.0.2.9-m9); automated positive cell counting was on three independent regions, with the diaminobezidine mean as the score compartment.24 (link)
McCann K., von Witzleben A., Thomas J., Wang C., Wood O., Singh D., Boukas K., Bendjama K., Silvestre N., Nielsen F.C., Thomas G., Sanchez-Elsner T., Greenbaum J., Schoenberger S., Peters B., Vijayanand P., Savelyeva N, & Ottensmeier C. (2022). Targeting the tumor mutanome for personalized vaccination in a TMB low non-small cell lung cancer. Journal for Immunotherapy of Cancer, 10(3), e003821.
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5

Immunohistochemistry Profiling of Tumor Markers

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Immunohistochemistry staining was carried out using mAbs against HSP105 (clone EPR4576; Epitomics), HLA class I (clone EMR8‐5; Hokudo), PD‐L1 (clone SP142; SPRING), CD8 (clone 1A5; BioGenex), and CD4 (clone 4B12; Dako) according to the manufacturers’ protocols. Quantitative analysis of HSP105 expression was carried out based on the staining intensity and the results were scored as 0, 1+, 2+, and 3+. The HLA class I expression was evaluated based on the proportion of stained cells and staining intensity; however, because all examined cancer tissues had a high proportion of stained cells (67% or more), they were classified based on the intensity. Expression of PD‐L1 was scored as – (0%) or + (1% or more) according to the percentage of positively stained areas relative to the total tumor areas. CD4+ and CD8+ T cells in tumors were counted in high power fields (×400) and the average numbers were calculated.
Shimizu Y., Yoshikawa T., Kojima T., Shoda K., Nosaka K., Mizuno S., Wada S., Fujimoto Y., Sasada T., Kohashi K., Bando H., Endo I, & Nakatsura T. (2019). Heat shock protein 105 peptide vaccine could induce antitumor immune reactions in a phase I clinical trial. Cancer Science, 110(10), 3049-3060.
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6

Immunohistochemical Analysis of Bone Marrow

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Immunohistochemical studies were carried out in 38 of 45 cases using sections of decalcified paraffin-embedded bone marrow biopsy specimens. The following antibodies were used at the dilutions suggested by the manufacturers: CD3 (polyclonal, Dako), CD4 (clone 4B12, Dako), CD8 (clone C8/144B, Dako), CD20 (clone L26, Dako), CD56 (clone 123C3, Dako), CD57 (clone TB01, Dako), granzyme B (clone GrB-7, Dako), and T-cell restricted intracellular antigen 1 (TIA-1) (clone 2G9, Immunotech, France). After dewaxing and heat-induced antigen retrieval, immunostaining was performed on an Autostainer Link 48 (Dako, Denmark) according to the manufacturer’s instructions. All immunostained samples were counterstained with hematoxylin.
Gorodetskiy V.R., Sidorova Y.V., Kupryshina N.A., Vasilyev V.I., Probatova N.A., Ryzhikova N.V, & Sudarikov A.B. (2020). Analysis of a single-institution cohort of patients with Felty's syndrome and T-cell large granular lymphocytic leukemia in the setting of rheumatoid arthritis. Rheumatology International, 41(1), 147-156.
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7

Immunohistochemical Analysis of Splenic Immune Cells

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Immunohistochemistry was performed on an FFPE spleen sample from patient #13. The following antibodies were used at dilutions suggested by the manufacturers: CD3 (polyclonal, Dako, Carpinteria, CA, USA); CD4 (clone 4B12, Dako); CD8 (clone C8/144B, Dako); CD16 (clone 2H7, Novocastra Laboratories, Newcastle upon Tyne, UK); CD20 (clone L26, Dako); and TIA-1 (clone 2G9, Immunotech, France). Immunostaining was performed using an Autostainer Link 48 (Dako, Denmark) according to the manufacturer’s instructions. All immunostained samples were counter-stained with hematoxylin.
Gorodetskiy V., Vasilyev V., Sidorova Y., Biderman B., Kupryshina N., Vagida M., Ryzhikova N, & Sudarikov A. (2022). Clinical Study of the Relationship between Sjögren Syndrome and T-Cell Large Granular Lymphocytic Leukemia: Single-Center Experience. International Journal of Molecular Sciences, 23(21), 13345.
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8

Immunohistochemical Analysis of Spleen and Bone Marrow

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Immunohistochemical studies of the spleen (5 patients) and bone marrow (15 patients) were conducted using the formalin-fixed paraffin-embedded tissue. The following antibodies were used at dilutions suggested by the manufacturers: CD3 (polyclonal, Dako, Carpinteria, CA, United States); CD4 (clone 4B12, Dako); CD8 (clone C8/144B, Dako); CD16 (clone 2H7, Novocastra Laboratories, Newcastle upon Tyne, United Kingdom); CD20 (clone L26, Dako); granzyme B (clone GrB-7, Dako); T-cell restricted intracellular antigen 1 (TIA-1) (clone 2G9, Immunotech, France); TCR-β F1 (clone 8A3, Thermo Scientific, Waltham, MA, United States); and TCR-γ (clone γ3.20, Thermo Scientific). After dewaxing and heat-induced antigen retrieval, immunostaining was performed using an Autostainer Link 48 (Dako, Denmark) according to the manufacturer’s instructions. All immunostained samples were counter-stained with hematoxylin.
Gorodetskiy V., Sidorova Y., Biderman B., Kupryshina N., Ryzhikova N, & Sudarikov A. (2022). STAT3 mutations in “gray-zone” cases of T-cell large granular lymphocytic leukemia associated with autoimmune rheumatic diseases. Frontiers in Medicine, 9, 1000265.
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9

Immunohistochemical Profiling of FFPE Skin Biopsies

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Skin biopsy specimens were routinely fixed in a 10% neutral-buffered formalin solution, and embedded in paraffin. The 4.5-µm sections of FFPE blocks were stained by H&E. Immunohistochemical analysis using the biotin-free alkaline phosphatase method (Leica Biosystems, Newcastle, UK) was performed using the fully automated stainer BOND-III systems (Leica Biosystems) with the following primary antibodies: CD3 (clone LN10, Leica Biosystems), CD4 (clone 4B12, DakoCytomation, Glostrup, Denmark), CD5 (clone 4C7, Leica Biosystems), CD20 (clone LR26, Leica Biosystems), CD56 (clone NCAM, Leica Biosystems), CD68 (clone 514H12, Leica Biosystems), CD123 (clone BR4MS, Novocastra, Leica Biosystems), TCL-1 (clone MRQ-7, Cell Marque, CA Rocklin, USA), TIA-1 (Biocare Medical, CA Pacheo, USA), and MPO (Cell Marque). Irrelevant IgG subclass‒matched antibodies were used for negative controls.
Shumilov E., Mazzeo P., Ghandili S., Künstner A., Weidemann S., Banz Y., Ströbel P., Pollak M., Kolloch L., Beltraminelli H., Kerkhoff A., Mikesch J.H., Schliemann C., Haase D., Wulf G., Legros M., Lenz G., Feldmeyer L., Pabst T., Witte H., Gebauer N, & Bacher U. (2024). Diagnostic management of blastic plasmacytoid dendritic cell neoplasm (BPDCN) in close interaction with therapeutic considerations. Annals of Hematology, 103(5), 1587-1599.
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10

Immunohistochemical Analysis of Tissue Samples

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Bone marrow biopsies were decalcified in EDTA prior to paraffin embedding. 3 μm sections were cut from formalin-fixed, paraffin-embedded samples. Upper gastrointestinal biopsies were routinely stained with hematoxylin and eosin stain and periodic acid–Schiff stain. Bone marrow biopsies were routinely stained with naphthol AS-D chloroacetate esterase (NASDCL). Immunohistochemical staining was performed using a horseradish peroxidase catalyzed brown chromogen reaction together with ready-to-use antibodies in an automated staining system (Dako Autostainer Link®; Dako, Glostrup, Denmark), following the manufacturer's guidelines. Depending on the tissue section size, up to three droplet zones are stained with 100 μl antibody solution per droplet zone. The following antibodies were used: CD3 (rabbit polyclonal, Dako), CD4 (clone 4B12, Dako), CD8 (clone C8/144B, Dako), CD19 (clone LE-CD19, Dako), CD20 (clone L26, Dako), CD38 (clone SPC 32, Novocastra, Newcastle upon Tyne, UK), IgM, IgG, IgA (rabbit polyclonal antibodies; Dako). Photos were taken on an Olympus BX51 microscope (Olympus Germany, Hamburg, Germany) with the AxioCam MRc camera (Zeiss, Jena, Germany).
Schubert D., Bode C., Kenefeck R., Hou T.Z., Wing J.B., Kennedy A., Bulashevska A., Petersen B.S., Schäffer A.A., Grüning B.A., Unger S., Frede N., Baumann U., Witte T., Schmidt R.E., Dueckers G., Niehues T., Seneviratne S., Kanariou M., Speckmann C., Ehl S., Rensing-Ehl A., Warnatz K., Rakhmanov M., Thimme R., Hasselblatt P., Emmerich F., Cathomen T., Backofen R., Fisch P., Seidl M., May A., Schmitt-Graeff A., Ikemizu S., Salzer U., Franke A., Sakaguchi S., Walker L.S., Sansom D.M, & Grimbacher B. (2014). Autosomal-dominant immune dysregulation syndrome in humans with CTLA4 mutations. Nature medicine, 20(12), 1410-1416.
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