STX17 - Antibodies - The Human Protein Atlas


	Antibody HPA001204

ANTIBODY INFORMATION
Provider	Atlas Antibodies Sigma-Aldrich
Product name	HPA001204
Host species	Rabbit
Clonalityⁱ The antibodies are designated mAB for monoclonal and pAb for polyclonal.	pAb
Concentration	0.27 mg/ml
Purity	Affinity purified using the PrEST-antigen as affinity ligand
Released in versionⁱ The release of the Human Protein Atlas in which the antibody was first published.	1.2
Referencesⁱ References to publications in which the antibody has been used.	21
Proper citation	Atlas Antibodies Cat#HPA001204, RRID:AB_1080118
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
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.	Uncertainⁱ 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 not consistent with literature. Immunofluorescent staining of human cell line U2OS shows localization to nucleoli and cytosol.

Antibody dilution	Human assay: A-431 fixed with PFA, dilution: 1:133 Human assay: U-251MG fixed with PFA, dilution: 1:133 Human assay: U2OS fixed with PFA, dilution: 1:133
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.	Approvedⁱ 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 adrenal gland shows strong cytoplasmic positivity in cortical cells. Adrenal gland

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
Antibody dilution	1:250
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.	Partly 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.	Low 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.	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. Band of predicted size in kDa (+/-20%) with additional bands present. Analysis performed using a standard panel of samples. 206 113 82 49 32 26 18

Antibody dilution	1:250
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.	Supported Pass with single peak corresponding to interaction only with its own antigen. Antibody specificity analysis with protein arrays. Predicted and matching interactions are shown in green.

Antibody dilution	1:3000
RELEVANT PUBLICATIONS
	The HOPS complex mediates autophagosome-lysosome fusion through interaction with syntaxin 17 Jiang P et al Mol Biol Cell 2014;25(8):1327-37 Application: ICC-IF, IP, WB
	ATG14 promotes membrane tethering and fusion of autophagosomes to endolysosomes Diao J et al Nature 2015;520(7548):563-6 Application: WB
	Impairment of autophagosome-lysosome fusion in the buff mutant mice with the VPS33A(D251E) mutation Zhen Y et al Autophagy 2015;11(9):1608-22 Application: IP, WB
	The Autophagosomal SNARE Protein Syntaxin 17 Is an Essential Factor for the Hepatitis C Virus Life Cycle Ren H et al J Virol 2016;90(13):5989-6000 Application: ICC-IF, WB
	Syntaxin-17 delivers PINK1/parkin-dependent mitochondrial vesicles to the endolysosomal system McLelland GL et al J Cell Biol 2016;214(3):275-91 Application: IP
	LAMP-2 is required for incorporating syntaxin-17 into autophagosomes and for their fusion with lysosomes Hubert V et al Biol Open 2016;5(10):1516-1529 Application: WB
	Dedicated SNAREs and specialized TRIM cargo receptors mediate secretory autophagy Kimura T et al EMBO J 2017;36(1):42-60 Application: WB
	Multiplex image-based autophagy RNAi screening identifies SMCR8 as ULK1 kinase activity and gene expression regulator Jung J et al Elife 2017;6: Application: IHC
	Transgenic expression of human APOL1 risk variants in podocytes induces kidney disease in mice Beckerman P et al Nat Med 2017;23(4):429-438 Application: ICC-IF
	Accumulation of undegraded autophagosomes by expression of dominant-negative STX17 (syntaxin 17) mutants Uematsu M et al Autophagy 2017;13(8):1452-1464 Application: ICC-IF, WB
	Protocol for Establishing a Multiplex Image-based Autophagy RNAi Screen in Cell Cultures Jung J et al Bio Protoc 2017;7(17):27982478 Application: ICC-IF
	Mechanism of Stx17 recruitment to autophagosomes via IRGM and mammalian Atg8 proteins Kumar S et al J Cell Biol 2018;217(3):997-1013 Application: WB
	A novel autophagy inhibitor berbamine blocks SNARE-mediated autophagosome-lysosome fusion through upregulation of BNIP3 Fu R et al Cell Death Dis 2018;9(2):243 Application: WB
	cPKCγ-Modulated Sequential Reactivation of mTOR Inhibited Autophagic Flux in Neurons Exposed to Oxygen Glucose Deprivation/Reperfusion Hua R et al Int J Mol Sci 2018;19(5): Application: ICC-IF
	Phosphorylation of ULK1 affects autophagosome fusion and links chaperone-mediated autophagy to macroautophagy Wang C et al Nat Commun 2018;9(1):3492 Application: IP, WB
	Autophagosomal YKT6 is required for fusion with lysosomes independently of syntaxin 17 Matsui T et al J Cell Biol 2018;217(8):2633-2645 Application: WB
	FIP200 Claw Domain Binding to p62 Promotes Autophagosome Formation at Ubiquitin Condensates Turco E et al Mol Cell 2019;74(2):330-346.e11 Application: WB
	An alternative mitophagy pathway mediated by Rab9 protects the heart against ischemia Saito T et al J Clin Invest 2019;129(2):802-819 Application: WB
	Syntaxin 17 promotes lipid droplet formation by regulating the distribution of acyl-CoA synthetase 3 Kimura H et al J Lipid Res 2018;59(5):805-819 Application: WB
	STX17 dynamically regulated by Fis1 induces mitophagy via hierarchical macroautophagic mechanism Xian H et al Nat Commun 2019;10(1):2059 Application: WB
	A reversible autophagy inhibitor blocks autophagosome-lysosome fusion by preventing Stx17 loading onto autophagosomes Vats S et al Mol Biol Cell 2019;30(17):2283-2295 Application: WB
ANTIGEN INFORMATION
Antigen	Recombinant protein fragment
Length (aa)	144
Antigen sequence	`RLEPAIQKFIKIVIPTDLERLRKHQINIEKYQRCRIWDKLHEEHINAGRT VQQLRSNIREIEKLCLKVRKDDLVLLKRMIDPVKEEASAATAEFLQLHLE SVEELKKQFNDEETLLQPPLTRSMTVGGAFHTTEAEASSQSLTQ`
Matching transcripts	STX17-201 - ENSP00000259400 [100%] STX17-205 - ENSP00000435981 [100%] STX17-212 - ENSP00000433484 [100%]
Matching mouse transcripts	ENSMUSP00000068087 [90%] ENSMUSP00000103348 [90%] ENSMUSP00000117512 [81%] ENSMUSP00000103349 [74%] ENSMUSP00000083899 [24%] ENSMUSP00000105712 [23%]
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.

STX17-201 STX17-205 STX17-212

Displaying protein features on the AlphaFold structures

The Protein Browser