SDHB - Antibodies - The Human Protein Atlas


	Antibody HPA002868	Antibody CAB009822	Antibody CAB068233	Antibody CAB068234	Antibody CAB068235

ANTIBODY INFORMATION
Provider	Atlas Antibodies Sigma-Aldrich	Santa Cruz Biotechnology	Atlas Antibodies	Atlas Antibodies	Atlas Antibodies
Product name	HPA002868	sc-25851	AMAb90705	AMAb90706	AMAb90708
Host species	Rabbit	Rabbit	Mouse	Mouse	Mouse
Clonalityⁱ The antibodies are designated mAB for monoclonal and pAb for polyclonal.	pAb	pAb	mAb	mAb	mAb
Concentration	0.0675 mg/ml	Not known	Not known	Not known	Not known
Purity	Affinity purified using the PrEST-antigen as affinity ligand	Protein A/G	Protein A/G	Protein A/G	Protein A/G
Released in versionⁱ The release of the Human Protein Atlas in which the antibody was first published.	3.1	4.1	13.0	13.0	13.0
Referencesⁱ References to publications in which the antibody has been used.	24
Proper citation	Atlas Antibodies Cat#HPA002868, RRID:AB_1079889	Santa Cruz Biotechnology Cat#sc-25851, RRID:AB_2183458	Atlas Antibodies Cat#AMAb90705, RRID:AB_2665638	Atlas Antibodies Cat#AMAb90706, RRID:AB_2665639	Atlas Antibodies Cat#AMAb90708, RRID:AB_2665640
Validation summaryⁱ All assays through which the antibody has been validated. Assays&annotation provide a detailed description of the different assays. The pie-charts indicate degree of validation.	ICC IHC WB PA	N/A ICC IHC WB N/A PA	N/A ICC IHC WB N/A PA	N/A ICC IHC WB N/A PA	N/A ICC IHC WB N/A PA
IMMUNOCYTOCHEMISTRYⁱ Immunocytochemistry is used to validate the antibody staining and for assessing and validating the protein expression pattern in selected human cell lines.
Validationⁱ Results of validation by standard or enhanced validation. Standard validation is based on concordance with available experimental gene/protein characterization data in the UniProtKB/Swiss-Prot database. Standard validation results in scores Supported, Approved or Uncertain. Enhanced validation is performed using either siRNA knockdown, tagged GFP cell lines or independent antibodies. For the siRNA validation the decrease in antibody-based staining intensity upon target protein downregulation is evaluated. For the GFP validation the signal overlap between the antibody staining and the GFP-tagged protein is evaluated. For the independent antibodies validation the evaluation is based on comparison of the staining of two (or more) independent antibodies directed towards independent epitopes on the protein. For all cases except the siRNA validation, an image representative of the antibody staining pattern is shown. For the siRNA validation, a box plot of the results is shown.	Supportedⁱ Reliability scores for antibodies used in immunocytochemistry are set by comparing the staining pattern in cell lines with external experimental evidence for protein localization. The scores are termed Supported, Approved and Uncertain. The subcellular location is supported by literature. Immunofluorescent staining of human cell line PC-3 shows localization to nucleoplasm, plasma membrane and mitochondria.	N/A	N/A	N/A	N/A

Antibody dilution	Human assay: A-431 fixed with PFA, dilution: 1:34 Human assay: PC-3 fixed with PFA, dilution: 1:34 Human assay: U2OS fixed with PFA, dilution: 1:34
IMMUNOHISTOCHEMISTRYⁱ Immunohistochemistry is used for validating antibody reliability by assessing staining pattern in 44 normal tissues. Validation scores include Enhanced, Supported, Approved and Uncertain.
Validationⁱ Results of validation by standard or enhanced validation based on assessment of antibody performance in 44 normal tissues. Standard validation results in scores Supported, Approved or Uncertain. An image representative of the antibody staining pattern is shown. Enhanced validation results in the score Enhanced and includes two methods: Orthogonal validation and Independent antibody validation. For orthogonal validation, representative images of high and low expression are shown. For independent antibody validation, four images of each independent antibody are displayed.	Enhanced - Orthogonal Antibody staining mainly consistent with RNA expression data across 43 tissues. HIGH EXPRESSION Heart muscle RNA expression: 293.0 nTPM LOW EXPRESSION Cervix RNA expression: 54.7 nTPM	Enhanced - Orthogonal Antibody staining mainly consistent with RNA expression data across 44 tissues. HIGH EXPRESSION Heart muscle RNA expression: 293.0 nTPM LOW EXPRESSION Cervix RNA expression: 54.7 nTPM	Supportedⁱ Immunohistochemistry is used for validating antibody reliability by assessing staining pattern in 44 normal tissues. Validation scores include Enhanced, Supported, Approved and Uncertain. Immunohistochemical staining of human liver shows strong positivity in hepatocytes. Liver	Supportedⁱ Immunohistochemistry is used for validating antibody reliability by assessing staining pattern in 44 normal tissues. Validation scores include Enhanced, Supported, Approved and Uncertain. Immunohistochemical staining of human liver shows strong granular cytoplasmic positivity in hepatocytes. Liver	Enhanced - Orthogonal Antibody staining mainly consistent with RNA expression data across 44 tissues. HIGH EXPRESSION Heart muscle RNA expression: 293.0 nTPM LOW EXPRESSION Cervix RNA expression: 54.7 nTPM

Retrievalⁱ Antigen retrieval is a method used to restore/retrieve the epitope (antibody bidning region) of the target protein, cross-linked, and thus masked, during tissue preserving fixative treatment of the tissues.	HIER pH6	HIER pH6	HIER pH6	HIER pH6	HIER pH6
Antibody dilution	1:250	1:100	1:500	1:500	1:1400
Literature conformityⁱ Conformance of the expression pattern with available gene/protein characterization data in scientific literature and data from bioinformatic predictions. UniProt is used as the main source of gene/protein characterization data and when relevant, available publications and other sources of information are researched in depth. Extensive or sufficient gene/protein data requires that there is evidence of existence on a protein level and that a substantial quantity of published experimental data is available from literature and public databases. Limited protein/gene characterization data does not require evidence of existence on a protein level and refers to genes for which only bioinformatic predictions and scarce published experimental data is available.	Consistent with extensive gene/protein characterization data.	Consistent with extensive gene/protein characterization data.	Consistent with extensive gene/protein characterization data.	Consistent with extensive gene/protein characterization data.	Consistent with extensive gene/protein characterization data.
RNA consistencyⁱ Consistency between immunohistochemistry data and consensus RNA levels is divided into five different categories: i) High consistency, ii) Medium consistency, iii) Low consistency, iv) Very low consistency, and v) Cannot be evaluated.	High consistency between antibody staining and RNA expression data.	High consistency between antibody staining and RNA expression data.	High consistency between antibody staining and RNA expression data.	High consistency between antibody staining and RNA expression data.	High consistency between antibody staining and RNA expression data.
WESTERN BLOTⁱ A Western blot analysis is performed on a panel of human tissues and cell lines to evaluate antibody specificity. For antibodies with unreliable result a revalidation using an over-expression lysate is performed.
Validationⁱ Western Blot is used for quality control of the polyclonal antibodies generated in the project. After purification, the antibodies are used to detect bands in a setup of lysate and different tissues. The result is then scored Enhanced, Supported, Approved, or Uncertain. Enhanced validation includes five different methods: Genetic validation, Recombinant expression validation, Independent antibody validation, Orthogonal validation and Capture MS validation.	Enhanced - Capture MSⁱ This method is based on comparison between the molecular weight of the stained band visualized by the antibody against the protein size obtained by a capture MS method in which multiple gel slices are cut out from the electrophoretic separation of cell lysates of RT4 and U-251 and analysed separately by proteomics. The band detected by the antibody should be equivalent to the same of the intended target protein and its peptide(s). Antibody band pattern is confirmed by capture-MS. 206 113 82 49 32 26 18	Supportedⁱ The staining of an antibody is evaluated by Western Blot through analysis of samples from different cell lysates. A supportive score is given if band(s) of predicted size in kDa (+/-20%) is detected. Single band corresponding to the predicted size in kDa (+/-20%). Analysis performed using a standard panel of samples. 219 111 83 48 32 26 17	Supportedⁱ The staining of an antibody is evaluated by Western Blot through analysis of samples from different cell lysates. A supportive score is given if band(s) of predicted size in kDa (+/-20%) is detected. Single band corresponding to the predicted size in kDa (+/-20%). Analysis performed using a standard panel of samples. 250 130 100 70 55 35 25 15 10	Supportedⁱ The staining of an antibody is evaluated by Western Blot through analysis of samples from different cell lysates. A supportive score is given if band(s) of predicted size in kDa (+/-20%) is detected. Single band corresponding to the predicted size in kDa (+/-20%). Analysis performed using a standard panel of samples. 250 130 100 70 55 35 25 15 10	Supportedⁱ The staining of an antibody is evaluated by Western Blot through analysis of samples from different cell lysates. A supportive score is given if band(s) of predicted size in kDa (+/-20%) is detected. Single band corresponding to the predicted size in kDa (+/-20%). Analysis performed using a standard panel of samples. 250 130 100 70 55 35 25 15 10

Antibody dilution	1:250	1:500	1:1000	1:1000	1:1000
PROTEIN ARRAY
Validationⁱ A protein array containing 384 different antigens including the antibody target is used to analyse antibody specificity. Depending on the array interaction profile the antibody is scored as Supported, Approved, or Uncertain.	Approved Pass with quality comment low specificity (binding to 1-2 antigens >15% and <40%). Antibody specificity analysis with protein arrays. Predicted and matching interactions are shown in green.	N/A	N/A	N/A	N/A

Antibody dilution	1:3000
RELEVANT PUBLICATIONS
	An immunohistochemical procedure to detect patients with paraganglioma and phaeochromocytoma with germline SDHB, SDHC, or SDHD gene mutations: a retrospective and prospective analysis van Nederveen FH et al Lancet Oncol 2009;10(8):764-71
	The Warburg effect is genetically determined in inherited pheochromocytomas Favier J et al PLoS One 2009;4(9):e7094
	SDHA is a tumor suppressor gene causing paraganglioma Burnichon N et al Hum Mol Genet 2010;19(15):3011-20
	A non-catecholamine-producing sympathetic paraganglioma of the spermatic cord: the importance of performing candidate gene mutation analysis Alataki D et al Virchows Arch 2010;457(5):619-22
	SDHB immunohistochemistry: a useful tool in the diagnosis of Carney-Stratakis and Carney triad gastrointestinal stromal tumors Gaal J et al Mod Pathol 2011;24(1):147-51
	Sumo1-ylation of human spermatozoa and its relationship with semen quality Marchiani S et al Int J Androl 2011;34(6 Pt 1):581-93
	Succinate dehydrogenase (SDH) D subunit (SDHD) inactivation in a growth-hormone-producing pituitary tumor: a new association for SDH? Xekouki P et al J Clin Endocrinol Metab 2012;97(3):E357-66
	Mitochondrial function and content in pheochromocytoma/paraganglioma of succinate dehydrogenase mutation carriers Rapizzi E et al Endocr Relat Cancer 2012;19(3):261-9
	Analysis of all subunits, SDHA, SDHB, SDHC, SDHD, of the succinate dehydrogenase complex in KIT/PDGFRA wild-type GIST Pantaleo MA et al Eur J Hum Genet 2014;22(1):32-9 Application: IHC
	A MEN1 syndrome with a paraganglioma Jamilloux Y et al Eur J Hum Genet 2014;22(2):283-5 Application: IHC
	Tyrosine kinase receptors as molecular targets in pheochromocytomas and paragangliomas Cassol CA et al Mod Pathol 2014;27(8):1050-62 Application: IHC
	Aberrant DNA hypermethylation of SDHC: a novel mechanism of tumor development in Carney triad Haller F et al Endocr Relat Cancer 2014;21(4):567-77 Application: WB
	Integrated genomic study of quadruple-WT GIST (KIT/PDGFRA/SDH/RAS pathway wild-type GIST) Nannini M et al BMC Cancer 2014;14:685 Application: IHC
	Paraganglioma and pheochromocytoma upon maternal transmission of SDHD mutations Bayley JP et al BMC Med Genet 2014;15:111 Application: IHC
	C11orf83, a mitochondrial cardiolipin-binding protein involved in bc1 complex assembly and supercomplex stabilization Desmurs M et al Mol Cell Biol 2015;35(7):1139-56 Application: WB
	Urinary Bladder Paragangliomas: How Immunohistochemistry Can Assist to Identify Patients With SDHB Germline and Somatic Mutations Giubellino A et al Am J Surg Pathol 2015;39(11):1488-92 Application: IHC
	Deciphering the molecular basis of invasiveness in Sdhb-deficient cells Loriot C et al Oncotarget 2015;6(32):32955-65 Application: WB
	NDUFAF5 Hydroxylates NDUFS7 at an Early Stage in the Assembly of Human Complex I Rhein VF et al J Biol Chem 2016;291(28):14851-60 Application: WB
	Hypoxia potentiates the cytotoxic effect of piperlongumine in pheochromocytoma models Bullova P et al Oncotarget 2016;7(26):40531-40545 Application: IP
	or - mutations do not predict outcome and do not cause loss of 5-hydroxymethylcytosine or altered histone modifications in central chondrosarcomas Cleven AHG et al Clin Sarcoma Res 2017;7:8 Application: IHC
	Validation of pathological grading systems for predicting metastatic potential in pheochromocytoma and paraganglioma Koh JM et al PLoS One 2017;12(11):e0187398 Application: IHC
	Metabolome-guided genomics to identify pathogenic variants in isocitrate dehydrogenase, fumarate hydratase, and succinate dehydrogenase genes in pheochromocytoma and paraganglioma Richter S et al Genet Med 2019;21(3):705-717 Application: IHC
	The loss of succinate dehydrogenase B expression is frequently identified in hemangioblastoma of the central nervous system Roh TH et al Sci Rep 2019;9(1):5873 Application: IHC
	Genetic Profile of Indian Pheochromocytoma and Paraganglioma Patients - A Single Institutional Study Agarwal G et al Indian J Endocrinol Metab ;23(4):486-490 Application: IHC
ANTIGEN INFORMATION
Antigen	Recombinant protein fragment	Recombinant protein	Recombinant protein	Recombinant protein	Recombinant protein
Length (aa)	109
Antigen sequence	`EGKQQYLQSIEEREKLDGLYECILCACCSTSCPSYWWNGDKYLGPAVLMQ AYRWMIDSRDDFTEERLAKLQDPFSLYRCHTIMNCTRTCPKGLNPGKAIA EIKKMMATY`
Matching transcripts	SDHB-201 - ENSP00000364649 [100%]
Matching mouse transcripts	ENSMUSP00000010007 [97%] ENSMUSP00000133917 [22%] ENSMUSP00000142386 [19%]
ANTIGEN VIEWⁱ The Structure section provides in-house generated structures, predicted using the Alphafold source code, for the majority of the proteins and their related isoforms. Displaying protein features on the AlphaFold structures Individual splice variants can be selected in the top part of the Protein Browser (see below) and different transcript-related features such as transmembrane regions, InterPro domains and antigen sequences for antibodies can be displayed in the structure by clicking on the respective features in the Protein Browser. Clinical and population-based amino acid variants based on data from the Ensembl variation database and AlphaMissense (AM) predictions can be highlighted using the sliders to the right of the structure. These can also be used to colour the entire structure by residue index or make the structure autorotate.The structures are displayed using the NGL Viewer and can also be zoomed-in and rotated manually. The Protein Browser The ProteinBrowser displays the antigen location on the target protein(s) and the features of the target protein. Transcript names and schematic transcript structures including exons, introns and UTRs for the different isoforms are shown on top, and can be used to switch between the structures for the different splice variants. At the top of the view, the position of the antigen (identified by the corresponding HPA identifier) is shown as a green bar. A yellow triangle on the bar indicates a <100% sequence identity to the protein target. Below the antigens, the maximum percent sequence identity of the protein to all other proteins from other human genes is displayed, using a sliding window of 10 aa residues (HsID 10) or 50 aa residues (HsID 50). The region with the lowest possible identity is always selected for antigen design, with a maximum identity of 60% allowed for designing a single-target antigen (read more). The curve in blue displays the predicted antigenicity i.e. the tendency for different regions of the protein to generate an immune response, with peak regions being predicted to be more antigenic.The curve shows average values based on a sliding window approach using an in-house propensity scale. (read more). Signal peptides (turquoise) and membrane regions (orange) based on predictions using the majority decision methods MDM and MDSEC are also displayed. Low complexity regions are shown in yellow and InterPro regions in green. Common (purple) and unique (grey) regions between different splice variants of the gene are also displayed (read more), and at the bottom of the protein view is the protein scale.

SDHB-201

Displaying protein features on the AlphaFold structures

The Protein Browser