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Hpa002868

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The HPA002868 is a laboratory equipment product manufactured by Merck Group. It is designed for use in various scientific research and analytical applications. The core function of this equipment is to perform specific tasks required in laboratory settings. However, a detailed and unbiased description of the product's features and capabilities is not available at this time.

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Lab products found in correlation

Ab14715, by Abcam (5 mentions) 2e3gc12fb2ae2, by Bio-Rad (2 mentions) Hpa027341, by Merck Group (2 mentions) Discovery xt system, by Roche (1 mentions) Ab155999, by Abcam (1 mentions) A5441, by Merck Group (1 mentions) Proteome profiler human phospho kinase array kit, by R&D Systems (1 mentions) Anti p62 ab56416, by Abcam (1 mentions) Pierce co immunoprecipitation kit, by Thermo Fisher Scientific (1 mentions) Bond polymer refine detection system, by Leica Biosystems (1 mentions) Ab95950, by Abcam (1 mentions) As48link, by Agilent Technologies (1 mentions)

13 protocols using hpa002868

1

SDHB Immunohistochemistry in Pheochromocytomas

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In addition to standard histological analysis, immunohistochemical analysis of SDHB protein was performed on paraffin-embedded tumors for all cases (polyclonal Anti-SDHB antibody, dilution 1/150, Sigma-Aldrich, HPA002868). Non-SDHx mutated PHEOs typically exhibit a granular cytoplasmic positive SDHB staining (mitochondrial pattern) whereas SDHx-related PHEOs are SDHB protein negative (normal tissue is used as positive control).
Maignan A., Guerin C., Julliard V., Paladino N.C., Kim E., Roche P., Castinetti F., Essamet W., Mancini J., Imperiale A., Clifton-Bligh R., Romanet P., Barlier A., Pacak K., Sebag F, & Taïeb D. (2017). Implications of SDHB genetic testing in patients with sporadic pheochromocytoma. Langenbeck's archives of surgery, 402(5), 787-798.
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2

Bladder Paraganglioma Molecular Characterization

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Paraganglioma tumor tissue was obtained from patients accepted for protocol evaluation at the National Institutes of Health (NIH), in accordance with the principles and procedures outlined in the NIH IRB Guidelines, and this was approved by the Institutional Review Board of the Eunice Kennedy Shriver National Institute of Child Health and Human Development (NICHD), NIH. All patients signed an IRB-approved consent that allowed for the collection of tissue samples.
Fourteen cases of urinary bladder paragangliomas were studied. Tumors were morphologically evaluated and stained for proliferative markers (MIB1), chromogranin and synaptophysin. Eleven cases were stained for SDHA and SDHB protein expression by IHC. One polyclonal antibodie (HPA002868, Sigma-Aldrich) was used to recognize of the target human protein for SDHB (recognizing the C-terminal).
Giubellino A., Lara K., Martucci V., Huynh T., Agarwal P., Pacak K, & Merino M.J. (2015). Urinary bladder paragangliomas: How Immunohistochemistry can assist to identify patients with SDHB germ line and somatic mutations. The American journal of surgical pathology, 39(11), 1488-1492.
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3

Immunohistochemical Detection of SDHB, SDHA, and FH

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FFPE tumor tissue was sectioned and stained for SDHB or FH protein using
rabbit polyclonal anti-SDHB (HPA002868, Sigma-Aldrich), anti-SDHA (ab14715,
Abcam or 2E3GC12FB2AE2, Biorad), and anti-FH (HPA027341, Sigma-Aldrich).
Richter S., Gieldon L., Pang Y., Peitzsch M., Huynh T., Leton R., Viana B., Ercolino T., Mangelis A., Rapizzi E., Menschikowski M., Aust D., Kroiss M., Beuschlein F., Gudziol V., Timmers H.J., Lenders J., Mannelli M., Cascon A., Pacak K., Robledo M., Eisenhofer G, & Klink B. (2018). Metabolome-guided genomics to identify mutations in isocitrate dehydrogenase, fumarate hydratase and succinate dehydrogenase genes in pheochromocytoma and paraganglioma. Genetics in medicine : official journal of the American College of Medical Genetics, 21(3), 705-717.
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4

Immunohistochemical Detection of SDHB, SDHA, and FH

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FFPE tumor tissue was sectioned and stained for SDHB or FH protein using
rabbit polyclonal anti-SDHB (HPA002868, Sigma-Aldrich), anti-SDHA (ab14715,
Abcam or 2E3GC12FB2AE2, Biorad), and anti-FH (HPA027341, Sigma-Aldrich).
Richter S., Gieldon L., Pang Y., Peitzsch M., Huynh T., Leton R., Viana B., Ercolino T., Mangelis A., Rapizzi E., Menschikowski M., Aust D., Kroiss M., Beuschlein F., Gudziol V., Timmers H.J., Lenders J., Mannelli M., Cascon A., Pacak K., Robledo M., Eisenhofer G, & Klink B. (2018). Metabolome-guided genomics to identify mutations in isocitrate dehydrogenase, fumarate hydratase and succinate dehydrogenase genes in pheochromocytoma and paraganglioma. Genetics in medicine : official journal of the American College of Medical Genetics, 21(3), 705-717.
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5

Immunohistochemical Assessment of SDHA and SDHB

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Immunohistochemistry for SDHA and SDHB proteins was performed as part of clinical assessment of tumors on formalin-fixed, paraffin-embedded tissue sections using AB14715 (Abcam, Cambridge MA, USA) and HPA002868 (Sigma–Aldrich, St. Louis, MO, USA) antibodies, respectively. Briefly, 4 μm thick sections from representative tissue blocks were processed using the Ventana Discovery XT system with antigen retrieval (CC1 solution, 60 min), primary antibody (1:6400 dilution for SDHA (AB14715) and 1:800 dilution for SDHB (HPA002868) antibodies), and OptiView DAB immunohistochemistry detection steps (Ventana Medical Systems, Tucson, AZ, USA).
Mandelker D., Marra A., Mehta N., Selenica P., Yelskaya Z., Yang C., Somar J., Mehine M., Misyura M., Basturk O., Latham A., Carlo M., Walsh M., Stadler Z.K., Offit K., Bandlamudi C., Hameed M., Chi P., Reis-Filho J.S, & Ceyhan-Birsoy O. (2023). Expanded genetic testing of GIST patients identifies high proportion of non-syndromic patients with germline alterations. NPJ Precision Oncology, 7, 1.
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6

Evaluating SDHB and SDHC Protein Levels

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Protein lysates were isolated from snap-frozen tumor and non-neoplastic tissues, and 10–50 μg of lysate were loaded on a western blot. Protein levels were evaluated for SDHB (1:500 dilution, HPA002868, Sigma–Aldrich) and SDHC (1:1000 dilution, ab155999, Abcam, Cambridge, UK), with ACTB (1:5000 dilution, A5441, Sigma–Aldrich) serving as loading control.
Haller F., Moskalev E.A., Faucz F.R., Barthelmeß S., Wiemann S., Bieg M., Assie G., Bertherat J., Schaefer I.M., Otto C., Rattenberry E., Maher E.R., Ströbel P., Werner M., Carney J.A., Hartmann A., Stratakis C.A, & Agaimy A. (2014). Aberrant DNA hypermethylation of SDHC: a novel mechanism of tumor development in Carney triad. Endocrine-related cancer, 21(4), 567-577.
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7

SDHB Immunohistochemistry in Tumours

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FFPE tissue was sectioned and stained for SDHB using rabbit polyclonal anti-SDHB (HPA002868, 1:400 dilution; Sigma-Aldrich, St Louis, MO, USA) according to the local procedures of six different centres (Dresden, Bethesda, Madrid, Florence, Nijmegen, and Rotterdam) [9 (link)]. For 23% of tumours (44 samples), a tissue microarray was constructed with three cores of 1.0 mm per sample. Local pathologists evaluated SDHB staining in one slide per tumour and gave the results in four categories: as positive, for the typical granular staining pattern; as negative, for completely negative or weak diffuse staining; as inconclusive, when both patterns were present; or as non-informative, when tissue or staining artefacts were observed. For 50 samples, the SDHB-IHC interpretations were as established previously, from combined interpretations of seven expert pathologists [20 (link)]; for these cases, results were rated as inconclusive when fewer than five pathologists agreed. All pathologists were blinded to the genetic status.
Wallace P.W., Conrad C., Brückmann S., Pang Y., Caleiras E., Murakami M., Korpershoek E., Zhuang Z., Rapizzi E., Kroiss M., Gudziol V., Timmers H.J., Mannelli M., Pietzsch J., Beuschlein F., Pacak K., Robledo M., Klink B., Peitzsch M., Gill A.J., Tischler A.S., de Krijger R.R., Papathomas T., Aust D., Eisenhofer G, & Richter S. (2020). Metabolomics, machine learning and immunohistochemistry to predict succinate dehydrogenase mutational status in phaeochromocytomas and paragangliomas. The Journal of pathology, 251(4), 378-387.
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8

Diagnosis of SDH-related Paraganglioma

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Sections from formalin-fixed paraffin embedded (FFPE) tumor blocks were stained with hematoxylin & eosin (H&E) using standard methods. An experienced neuroendocrine pathologist (RdK) evaluated slides and provided or confirmed (patient 3) the histopathological diagnosis. A recent and important development in the diagnosis of SDH-related paraganglioma has been the application of SDHB immunohistochemistry, which can reliably identify and differentiate SDH-related tumors from tumors with other causes regardless of which of the SDH genes is actually affected by a mutation [16 (link)]. SDHB immunohistochemistry was carried out as described [16 (link)], using a rabbit polyclonal primary SDHB antibody, HPA002868 (Sigma-Aldrich Corp; St Louis, MO, USA; 1:500).
Bayley J.P., Oldenburg R.A., Nuk J., Hoekstra A.S., van der Meer C.A., Korpershoek E., McGillivray B., Corssmit E.P., Dinjens W.N., de Krijger R.R., Devilee P., Jansen J.C, & Hes F.J. (2014). Paraganglioma and pheochromocytoma upon maternal transmission of SDHD mutations. BMC Medical Genetics, 15, 111.
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9

Western Blot and Immunoprecipitation Assay for Cell Signaling

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The western blotting was performed as reported previously [37 (link)]. RIP3 immunoprecipitation was performed using the Pierce Co-Immunoprecipitation kit according to the manufacturer's instructions (Thermo Scientific, Inc., Pittsburgh, PA, USA). The primary antibodies included anti-cleaved caspase 3 (5A1E; 9664), anti-caspase 3 (8G10; 9665), anti-cleaved PARP (9544), anti-LC3A/B (4108), anti-LAMP1 (C54H11; 3243), anti-γH2A.X (2577), anti-RIP1 (D94C12; 3493), anti-histone 3 (D1H2; 4499; Cell Signaling Technology, Danvers, MA, USA), anti-p62 (ab56416, Abcam, Cambridge, UK), anti-ubiquitin (FK2; ST1200; Millipore, Billerica, MA, USA), anti-RIP3, anti-β-tubulin (D66; T0198), anti-RIP1 (334640; MAB3585, R&D Systems, Minneapolis, MN, USA), anti-RIP3 (R4277) and anti-SDHB (HPA002868; Sigma-Aldrich, St. Louis, MO, USA). For the detection of phospo-kinases, the Proteome Profiler Human Phospho-Kinase Array Kit (R&D Systems, Inc, Minneapolis, MN, USA) was used and the samples were analyzed according to the manufacturer's instructions.
Bullova P., Cougnoux A., Abunimer L., Kopacek J., Pastorekova S, & Pacak K. (2016). Hypoxia potentiates the cytotoxic effect of piperlongumine in pheochromocytoma models. Oncotarget, 7(26), 40531-40545.
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10

SDHB Immunohistochemistry on FFPE Tissue

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SDHB immunohistochemistry was performed on 3-μm sections of FFPE tissue mounted on adhesive slides. The staining was performed on a fully automated BOND III IHC and ISH stainer system (Leica Biosystems, Nassloch, Germany). The SDHB primary antibody rabbit polyclonal (HPA002868, Sigma Aldrich, St Louis, MO, USA) was used. The Primary antibody binding to tissue sections was visualized using BOND Polymer Refine Detection system (DS9800, Leica Biosystems, Nassloch, Germany).
Whitworth J., Casey R.T., Smith P.S., Giger O., Martin J.E., Clark G., Cook J., Fernando M.S., Taniere P, & Maher E.R. (2021). Familial wild-type gastrointestinal stromal tumour in association with germline truncating variants in both SDHA and PALB2. European Journal of Human Genetics, 29(7), 1139-1145.
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