SATB2 - Antibodies - The Human Protein Atlas


	Antibody HPA001042	Antibody HPA029543	Antibody CAB023669	Antibody CAB062562	Antibody CAB067751	Antibody CAB068229	Antibody CAB068230	Antibody CAB068231

ANTIBODY INFORMATION
Provider	Atlas Antibodies Sigma-Aldrich	Atlas Antibodies Sigma-Aldrich	Santa Cruz Biotechnology	Atlas Antibodies	Atlas Antibodies	Atlas Antibodies	Atlas Antibodies	Atlas Antibodies
Product name	HPA001042	HPA029543	sc-81376	AMAb90635	AMAb90679	AMAb90678	AMAb90680	AMAb90682
Host species	Rabbit	Rabbit	Mouse	Mouse	Mouse	Mouse	Mouse	Mouse
Clonalityⁱ The antibodies are designated mAB for monoclonal and pAb for polyclonal.	pAb	pAb	mAb	mAb	mAb	mAb	mAb	mAb
Concentration	0.05 mg/ml	0.0412 mg/ml	Not known	Not known	Not known	Not known	Not known	Not known
Purity	Affinity purified using the PrEST-antigen as affinity ligand	Affinity purified using the PrEST-antigen as affinity ligand	Protein A/G	Protein A/G	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.	6.0	9.0	9.0	12.0	13.0	13.0	13.0	13.0
Referencesⁱ References to publications in which the antibody has been used.	4	1		4	2			1
Proper citation	Atlas Antibodies Cat#HPA001042, RRID:AB_10601711	Atlas Antibodies Cat#HPA029543, RRID:AB_10959737	Santa Cruz Biotechnology Cat#sc-81376, RRID:AB_1129287	Atlas Antibodies Cat#AMAb90635, RRID:AB_2665615	Atlas Antibodies Cat#AMAb90679, RRID:AB_2665629	Atlas Antibodies Cat#AMAb90678, RRID:AB_2665628	Atlas Antibodies Cat#AMAb90680, RRID:AB_2665630	Atlas Antibodies Cat#AMAb90682, RRID:AB_2665631
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.	N/A ICC IHC WB PA	ICC IHC WB PA	N/A ICC IHC WB N/A PA	N/A ICC IHC N/A 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	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.	N/A	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 U2OS shows localization to nucleoplasm.	N/A	N/A	N/A	N/A	N/A	N/A

Antibody dilution		Human assay: A-431 fixed with PFA, dilution: 1:20 Human assay: U-251MG fixed with PFA, dilution: 1:20 Human assay: U2OS fixed with PFA, dilution: 1:20
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 44 tissues. HIGH EXPRESSION Colon RNA expression: 27.2 nTPM LOW EXPRESSION Liver RNA expression: 2.9 nTPM Enhanced - Independent antibodies Protein distribution across 45 tissues similar between the independent antibodies HPA001042 and HPA029543. Cerebral cortex Colon Liver Rectum	Enhanced - Orthogonal Antibody staining mainly consistent with RNA expression data across 43 tissues. HIGH EXPRESSION Colon RNA expression: 27.2 nTPM LOW EXPRESSION Esophagus RNA expression: 0.8 nTPM Enhanced - Independent antibodies Protein distribution across 45 tissues similar between the independent antibodies HPA001042 and HPA029543. Cerebral cortex Colon Liver Rectum	Enhanced - Orthogonal Antibody staining mainly consistent with RNA expression data across 44 tissues. HIGH EXPRESSION Rectum RNA expression: 31.7 nTPM LOW EXPRESSION Stomach 1 RNA expression: 1.0 nTPM	Enhanced - Orthogonal Antibody staining mainly consistent with RNA expression data across 44 tissues. HIGH EXPRESSION Colon RNA expression: 27.2 nTPM LOW EXPRESSION Liver RNA expression: 2.9 nTPM	Enhanced - Orthogonal Antibody staining mainly consistent with RNA expression data across 44 tissues. HIGH EXPRESSION Rectum RNA expression: 31.7 nTPM LOW EXPRESSION Stomach 1 RNA expression: 1.0 nTPM	Enhanced - Orthogonal Antibody staining mainly consistent with RNA expression data across 44 tissues. HIGH EXPRESSION Colon RNA expression: 27.2 nTPM LOW EXPRESSION Liver RNA expression: 2.9 nTPM	Enhanced - Orthogonal Antibody staining mainly consistent with RNA expression data across 44 tissues. HIGH EXPRESSION Rectum RNA expression: 31.7 nTPM LOW EXPRESSION Stomach 1 RNA expression: 1.0 nTPM	Enhanced - Orthogonal Antibody staining mainly consistent with RNA expression data across 44 tissues. HIGH EXPRESSION Rectum RNA expression: 31.7 nTPM LOW EXPRESSION Stomach 1 RNA expression: 1.0 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	HIER pH6	HIER pH6	HIER pH6
Antibody dilution	1:50	1:500	1:100	1:300	1:1000	1:500	1:250	1:1000
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.	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.	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.	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. 219 112 85 49 32 25 18	Uncertainⁱ 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. No bands detected. Analysis performed using a standard panel of samples. 230 130 95 72 56 36 28 17 11	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 95 72 55 36 28 17 11	Uncertainⁱ 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. Weak band of predicted size but with additional bands of higher intensity also present. Analysis performed using a standard panel of samples.	Uncertainⁱ 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. No bands detected. Analysis performed using a standard panel of samples. 250 130 100 70 55 35 25 15 10	Uncertainⁱ 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. No bands detected. Analysis performed using a standard panel of samples. 250 130 100 70 55 35 25 15 10	Uncertainⁱ 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. No bands detected. 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	1:500	1:250	1:500	1:500	1:1000	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.	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.	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.	N/A	N/A	N/A	N/A	N/A	N/A

Antibody dilution	1:2000	1:500
RELEVANT PUBLICATIONS
	From gene expression analysis to tissue microarrays: a rational approach to identify therapeutic and diagnostic targets in lymphoid malignancies Ek S et al Mol Cell Proteomics 2006;5(6):1072-81	Molecular correlates and prognostic significance of SATB1 expression in colorectal cancer Nodin B et al Diagn Pathol 2012;7:115		Epitope mapping of antibodies using bacterial surface display Rockberg J et al Nat Methods 2008;5(12):1039-45	Prognostic and treatment predictive significance of SATB1 and SATB2 expression in pancreatic and periampullary adenocarcinoma Elebro J et al J Transl Med 2014;12:289 Application: IHC			Satb2 determines miRNA expression and long-term memory in the adult central nervous system Jaitner C et al Elife 2016;5: Application: ChIP
	Extensive expression of craniofacial related homeobox genes in canine mammary sarcomas Wensman H et al Breast Cancer Res Treat 2009;118(2):333-43			A cohort study of the prognostic and treatment predictive value of SATB2 expression in colorectal cancer Eberhard J et al Br J Cancer 2012;106(5):931-8	SATB1 is an independent prognostic factor in radically resected upper gastrointestinal tract adenocarcinoma Hedner C et al Virchows Arch 2014;465(6):649-59 Application: IHC, WB
	SATB2 in combination with cytokeratin 20 identifies over 95% of all colorectal carcinomas Magnusson K et al Am J Surg Pathol 2011;35(7):937-48			Scalable in situ hybridization on tissue arrays for validation of novel cancer and tissue-specific biomarkers Kiflemariam S et al PLoS One 2012;7(3):e32927
	Generation of monospecific antibodies based on affinity capture of polyclonal antibodies Hjelm B et al Protein Sci 2011;20(11):1824-35			Evaluating real-time immunohistochemistry on multiple tissue samples, multiple targets and multiple antibody labeling methods Dubois L et al BMC Res Notes 2013;6:542
ANTIGEN INFORMATION
Antigen	Recombinant protein fragment	Recombinant protein fragment	Recombinant protein	Recombinant protein	Recombinant protein	Recombinant protein	Recombinant protein	Recombinant protein
Length (aa)	123	115
Antigen sequence	`VSQAVFARVAFNRTQGLLSEILRKEEDPRTASQSLLVNLRAMQNFLNLPE VERDRIYQDERERSMNPNVSMVSSASSSPSSSRTPQAKTSTPTTDLPIKV DGANINITAAIYDEIQQEMKRAK`	`KECPLSQSMISSIVNSTYYANVSATKCQEFGRWYKKYKKIKVERVERENL SDYCVLGQRPMHLPNMNQLASLGKTNEQSPHSQIHHSTPIRNQVPALQPI MSPGLLSPQLSPQLV`
Matching transcripts	SATB2-201 - ENSP00000260926 [100%] SATB2-202 - ENSP00000401112 [100%] SATB2-203 - ENSP00000388581 [100%] SATB2-205 - ENSP00000388764 [100%] SATB2-206 - ENSP00000405420 [100%] SATB2-211 - ENSP00000514853 [100%] SATB2-213 - ENSP00000514854 [100%] SATB2-218 - ENSP00000514861 [100%]	SATB2-201 - ENSP00000260926 [100%] SATB2-202 - ENSP00000401112 [100%] SATB2-205 - ENSP00000388764 [100%] SATB2-206 - ENSP00000405420 [100%] SATB2-213 - ENSP00000514854 [100%] SATB2-218 - ENSP00000514861 [100%]
Matching mouse transcripts	ENSMUSP00000046067 [99%] ENSMUSP00000110057 [99%] ENSMUSP00000135391 [99%] ENSMUSP00000116006 [72%] ENSMUSP00000072904 [24%]	ENSMUSP00000046067 [100%] ENSMUSP00000110057 [100%] ENSMUSP00000135391 [66%] ENSMUSP00000128841 [54%] ENSMUSP00000036482 [20%]
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.

SATB2-201 SATB2-202 SATB2-203 SATB2-205 SATB2-206 SATB2-211 SATB2-213 SATB2-218

Displaying protein features on the AlphaFold structures

The Protein Browser