Clone kp1

Manufactured by Agilent Technologies

Sourced in Denmark, Japan

The Clone KP1 is a laboratory equipment product from Agilent Technologies. It is designed to perform cloning and protein expression tasks. The core function of the Clone KP1 is to facilitate the cloning and expression of DNA sequences in a controlled and efficient manner.

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CD68 (clone KP1, (20 citations) Anti-human CD68 (clone KP1, (5 citations) Mouse anti-CD68 (clone KP1, (4 citations) Anti‐CD68 antibody (clone KP1, (2 citations) Anti-CD68 FITC (clone KP1, (2 citations)

19 protocols using clone kp1

Neuropathological Characterization of Meningioma

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Neuropathological classification was performed based on the 2016 WHO criteria [3 (link)]. All histopathological reports underwent repeated examination to confirm that diagnosis was in line with these requirements. Immunohistochemistry was performed in the same workflow as described before for paraffin-embedded biopsy tissue specimen [31 (link),32 (link)]. The MIB-1 labeling index was investigated using the following antibody: Anti-Ki67 (Clone 2B11 + PD7/26). Moreover, semiquantitative analysis and scoring of the CD68⁺ stainings using anti-CD68 antibodies to detect macrophages were performed (Clone KP1, dilution 1:1000, DAKO, Glostrup, Denmark). As previously described, meningioma specimens were investigated for the absence, focal or diffuse staining of CD68⁺ macrophages [33 (link)]. Visualization was performed using diaminobenzidine and a neuropathological examination was performed by expert neuropathologists including A.J.B.

Wach J., Lampmann T., Güresir Á., Vatter H., Herrlinger U., Becker A., Toma M., Hölzel M, & Güresir E. (2022). Increased MIB-1 Labeling Index Is Associated with Abducens Nerve Morbidity in Primary Sporadic Petroclival Meningioma Surgery: Beyond Location and Approach. Current Oncology, 29(7), 5026-5041.

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Multiplex Immunohistochemistry for Tumor Immune Profiling

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ngTMAs were sectioned at 3 μm, dewaxed and rehydrated in distilled H₂O. They were double stained immunohistochemically for pancytokeratin (1:400, cytokeratin LMW, clone AE1/AE3, M3515, Dako-Agilent, Santa Clara, CA, USA) and each of the following: CD3 (1:400, clone SP7, ab16669, Abcam, Cambridge, UK), CD4 (1:100, clone CD4/4B12, M7310, Dako), CD8 (1:100, clone C8/144B, M7103, Dako), CD20 (1:100, clone L26, M0755, Dako), CD68 (1:100, clone KP1, M0814, Dako), DC-LAMP (1:100, CD208/DC-LAMP PA, 10527-H08H, Sino Biological, Beijing, China), iNOS (1:100, PAb, PA3-030A, Thermo Fisher Scientific, Waltham, MA, USA), CD163 (1:100, clone 10D6, NCL-CD163, Leica Biosystems AG, Muttenz, Switzerland) and FOXP3 (1:100, clone 236A/E7, ab20034, Abcam). Antigen retrieval was performed with Tris-HCl, pH 9 for 30 min at 95 °C. Antibody testing and staining protocols have been established and staining was performed by an automated Leica Bond RX System (Leica Bond RX, Leica Biosystems, Muttenz, Switzerland) with the Bond Polymer Refine Kit (with DAB as chromogen) and Bond Polymer Refine Red Detection Kit for the double staining (Leica Biosystems, Newcastle, UK).

Sadozai H., Acharjee A., Gruber T., Gloor B, & Karamitopoulou E. (2021). Pancreatic Cancers with High Grade Tumor Budding Exhibit Hallmarks of Diminished Anti-Tumor Immunity. Cancers, 13(5), 1090.

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Immunohistochemical Analysis of Carotid Plaques

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The carotid plaques were analyzed by immunohistochemistry. Paraformaldehyde-fixed sections of the specimens were embedded in paraffin. Serial sections were prepared on silanecoated glass slides, and they were immunostained with CD68 (1:20, mouse monoclonal, Clone JC70 A; DAKO, Glostrup, Denmark), CD117 (1:20, mouse monoclonal, Clone KP1; DAKO), CD4, and CD8 antibodies using a BOND-MAX autostainer (Leica Microsystems, Buffalo Grove, IL, USA). Images were acquired using a Keyence BZ-9000 microscope (Keyence Co., Osaka, Japan). Image J software (National Institutes of Health, Bethesda, MD, USA) was used for the semiquantitative analysis of the percentages of CD68-, CD117-, CD4-, and CD8-stained cells on the basis of the stained area. Histopathological analysis was performed by an experienced pathologist who was blinded to the patient characteristics and imaging findings.

Joo S.P., Lee S.W., Cho Y.H., Kim Y.S., Seo B.R., Kim H.S, & Kim T.S. (2019). Vasa Vasorum Densities in Human Carotid Atherosclerosis Is Associated with Plaque Development and Vulnerability. Journal of Korean Neurosurgical Society, 63(2), 178-187.

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Prostate Tumor Immune Infiltrates Characterization

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To characterize prominent prostate tumor infiltrates, we focused on abundant immunologic cell sub-types. We cut, stained and imaged archived formalin-fixed paraffin-embedded (FFPE) tumor tissue samples from the SCORE cases. Then we analyzed their cell type and count in the Department of Pathology and Laboratory Medicine at the University of Pennsylvania. Prostate sections from resected glands were stained for cytokeratin from tumor cells and T cells (CD3, CD8, FoxP3) and macrophages (CD68.) (Fig 1) The antibody clones that we used were CD3: clone LN 10 (Leica, catalog #NCL-L-3); CD8: clone C8/144B (DAKO, catalog #M7103); FOXP3: clone 206D (Biolegend, catalog #320102); and CD68: clone KP1 (DAKO, catalog #IR60961). The entire tumor nodule was scanned at low power to survey overall prevalence of infiltrating immune cells. We then selected 4 representative fields from each tumor sample. The area of the tumor that was selected depended on where the majority of cells were found in each tumor sample. Tissues were scored for infiltrating lymphocytes and macrophages in the dominant nodule (largest tumor nodule identified in the prostate) [21 (link)] by manually counting the number of TILs or TAMs in 4 fields at 20X for T cells and macrophages and averaging the scores.

Zeigler-Johnson C., Morales K.H., Lal P, & Feldman M. (2016). The Relationship between Obesity, Prostate Tumor Infiltrating Lymphocytes and Macrophages, and Biochemical Failure. PLoS ONE, 11(8), e0159109.

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Histopathological Grading and Macrophage Analysis in Meningioma

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Histopathological grading was performed based on the 2016 WHO criteria [3 (link)]. All histopathological reports underwent repeated review to reconfirm that the diagnosis was in keeping with these requirements. Immunohistochemistry was performed in a similar workflow, as described before for paraffin-embedded biopsy tissue specimen [32 (link),33 (link)]. The MIB-1 labeling index was determined by the usage of the following antibody: Anti-Ki67 (Clone 2B11 + PD7/26). Moreover, semiquantitative analysis and the scoring of CD68⁺ stainings using anti-CD68 antibodies to detect macrophages was performed (Clone KP1, dilution 1:1000, DAKO, Glostrup, Denmark). Meningioma specimens were investigated for the absence, focal or diffuse staining of CD68⁺ macrophages [34 (link)]. Visualization was by diaminobenzidine and the neuropathological assessment was performed by expert neuropathologists, including A.J.B. The further neuropathological workflows were as previously reported [34 (link)].

Wach J., Lampmann T., Güresir Á., Vatter H., Herrlinger U., Becker A., Toma M., Hölzel M, & Güresir E. (2022). Inflammatory Tumor Microenvironment in Cranial Meningiomas: Clinical Implications and Intraindividual Reproducibility. Diagnostics, 12(4), 853.

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Histological Analysis of Breast Implant Capsules

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Tissue samples were fixed in 10 % neutral-buffered formalin, then processed and embedded in paraffin. Sections were cut at 5 μm for hematoxylin and eosin (Richard-Allan Scientific, Kalamazoo, MI, USA) staining and immunohistochemistry.
Immunohistochemical evaluation was performed using monoclonal antibodies specific for α-smooth muscle actin (α-SMA), an indicator of myofibroblast presence (Clone 1A4, DAKO, Glostrup, Denmark) and for CD68 (Clone KP1, DAKO, Glostrup, Denmark [antibody recognizes a 110-kDa glycoprotein expressed on monocytes and macrophages]). All immunohistochemistry was performed using the EnVision™ FLEX High pH visualization system (DAKO, Glostrup, Denmark).
General characteristics of the histopathology of implant capsules with different Baker scores were assessed visually by review of hematoxylin and eosin-stained capsule samples. Capsules were classified into four categories: (1) dense collagen, acellular or low cellular content (example Fig. 4a), (2) dense collagen, moderate to high cellular content (example Fig. 4b), (3) synovial metaplasia (example Fig. 4c, d), or (4) loosely packed collagen (example Fig. 4e, f).

Bui J.M., Perry T., Ren C.D., Nofrey B., Teitelbaum S, & Van Epps D.E. (2015). Histological Characterization of Human Breast Implant Capsules. Aesthetic Plastic Surgery, 39(3), 306-315.

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Neuropathology of TUBB4A-related Hypomyelination

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A patient with severe hypomyelination and clinically, a profound lack of development, died at the age of almost 4 years because of respiratory failure. This patient harbored a de novo TUBB4A gene mutation, c.1242C>G / p.Asn414Lys, not present in the parents, in line with dominant de novo inheritance.
Post-mortem tissue was formalin-fixed paraffin-embedded for (immuno)histochemistry. Four-μm-thick tissue sections were stained with H&E, Klüver stain for myelin and Bodian stain for axons according to standard methods. After heat-induced antigen retrieval in 0.01M citrate buffer (pH6), immunohistochemical staining was performed with antibodies against the major myelin protein proteolipid protein (PLP; AbDserotec, 1:3000), the astrocyte-specific protein glial fibrillary acidic protein (GFAP; Millipore, 1:1000), and CD68 for microglia (Dako, clone KP1, 1:400). Immunoreactivity was detected with 3,30-diaminobenzidine as a chromogen. Tissue sections were photographed using a Leica DM6000B microscope (Leica Microsystems). Omitting primary antibodies yielded no significant staining.
EM was performed on frontal white matter. The tissue was fixed with 2% glutaraldehyde in 0.1M sodium cacodylate buffer (pH 7.4), post-fixed in 2% osmium tetroxide and embedded in epoxy resin. Ultrathin sections were stained with uranyl acetate and lead citrate.

Duncan I.D., Bugiani M., Radcliff A.B., Moran J.J., Lopez-Anido C., Duong P., August B.K., Wolf N.I., van der Knaap M.S, & Svaren J. (2017). A mutation in the Tubb4a gene leads to microtubule accumulation with hypomyelination and demyelination. Annals of neurology, 81(5), 690-702.

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Immunohistochemical Markers for Characterization

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Immunostains were performed on all cases and controls using steam antigen retrieval and standard techniques. Immunostains were performed for Keratin 7 (1:100; Clone OV-TL 12/30; Dako; CA), CD68 (1: 1500; Clone KP1; Dako, CA), Arginase (predilute; Clone SP156; Cell Marque, CA) and PRKAR1A (1:2000; Clone OTi6C7; Origene, MD).

Graham R.P., Lackner K., Terracciano L., González-Cantú Y., Maleszewski J.J., Greipp P.T., Simon S.M, & Torbenson M.S. (2018). Fibrolamellar Carcinoma in the Carney Complex: PRKAR1A Loss Instead of the Classic DNAJB1-PRKACA Fusion. Hepatology (Baltimore, Md.), 68(4), 1441-1447.

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Immunohistochemical Analysis of Tumor Proliferation

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Surgical specimens were routinely evaluated with haematoxylin/eosin staining and processed for immunohistochemical reactions with antibodies directed against: S-100 (DAKO, Glastrop, Denmark), Ki-67 (MIB1; DAKO, Glastrop, Denmark), and cluster of differentiation (CD) 68 (Clone KP1, dilution 1:1000, DAKO, Glastrop, Denmark). The MIB-1 index was determined in randomly selected high-power microscopic fields in 83 (83/87; 95.4%) patients. The proportions of stained and unstained nuclei in the tumor cells were determined. The MIB-1 index was defined as the percentage of Ki-67⁺ nuclei. Macrophage infiltrates were investigated using CD68 staining.

Wach J., Güresir Á., Borger V., Schuss P., Becker A., Coch C., Schmitz M.T., Hölzel M., Toma M., Herrlinger U., Vatter H, & Güresir E. (2021). Elevated baseline C-reactive protein levels predict poor progression-free survival in sporadic vestibular schwannoma. Journal of Neuro-Oncology, 156(2), 365-375.

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Cytological Evaluation of Papillary Thyroid Carcinoma

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A 30-year (1980-2013) retrospective review of the cytology slides from our cytology division of pathology department revealed a total of 750 PTCs. Out of these, 73 cases of CPTC, which were confirmed by histologic sections, were retrieved from the files: 55 females and 18 males, with an age range of 28-52 years. There was no pediatric case. The cyst sizes ranged between 2.8 cm and 3 cm. Aspiration was performed from the cysts tissue (42 cases under ultrasound guidance and the remaining without guidance). The cysts yielded 2-3 mL of the fluid that was hemorrhagic in 40 cases and clear yellow in color in the remaining cases. The smears were prepared from the sediment of cytocentrifuged specimens. The air-dried smears were stained with the Wright–Giemsa stain, and alcohol-fixed smears were stained with Papanicolaou stain. Some selected cases were subjected to immunocytochemistry for thyroid transcription factor-1 (TTF-1, Dako, clone 8G7G3/1, Denmark), cytokeratin 19 (CK 19) (Dako, Clone RCK108, Denmark), and cluster of differentiation 68 (CD68) (Dako, clone KP1, Denmark).
The FNA smears of other cystic thyroid lesions were retrieved from the files for the comparative study. These cases included 300 colloid goiters, 290 adenomatoid nodules, 11 follicular neoplasms, and 9 hurtle cell neoplasms.
Medical ethics committee of our university approved the study.

Mokhtari M., Kumar P.V, & Hayati K. (2016). Fine-needle aspiration study of cystic papillary thyroid carcinoma: Rare cytological findings. Journal of Cytology / Indian Academy of Cytologists, 33(3), 120-124.

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