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SATB1 / DNA-binding protein SATB1 · IHC design guide
Design Immunohistochemistry for SATB1
Plan SATB1 chromogenic IHC in paraffin sections using the expected nuclear pattern (HPA tissue IHC). Use thymic cortical cells or lymph node non-germinal center cells as positive references, and score staining by cell type (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
SATB1 Immunohistochemistry Experimental Design Guide
Expected localisation, validated protocols, controls and antibodies — the at-a-glance facts below, then the full design guide.
Must know before staining
| Expected localisation | Nuclear staining (HPA tissue IHC) | |
| Staining pattern | Nuclei of CNS and immune cells (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet M01312) | |
| Positive control | Cerebral cortex+4 more · see all | |
| Negative control | Adipose tissue+4 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep fixation consistent across sections (standard IHC practice; not target-specific) | |
| Caveat | Lymphoid staining is strongest in non-germinal center cells (HPA tissue IHC) | |
| Regulation | Predominantly expressed in thymus (UniProt) | |
| Isoform / epitope | Two isoforms; verify antibody epitope coverage (UniProt) |
Section 1
Recommended SATB1 IHC & IF Protocols
The catalog antibody protocol (datasheet: M01312) is followed by four published SATB1 IHC protocols (PMC3472180; PMC4023980; PMC5228311; PMC3523011).
| Sample | Paraffin-embedded human spleen tissue; fixative not specified (datasheet M01312) |
| Fixation | Image fixative and duration unreported (datasheet M01312); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat retrieval: EDTA pH 8.0 (datasheet M01312); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet M01312) |
| Primary antibody | Rabbit monoclonal (clone ACO-19) anti-SATB1, 1:50 (datasheet M01312) |
| Primary incubation | Overnight at 4 °C (datasheet M01312) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet M01312) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | SATB1-positive staining in neuronal cells of cerebral cortex (HPA tissue IHC: High). HPA tissue profile: Nuclear expression in several tissues, mainly in cells of the CNS and immune system. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA retrieval at pH 8.0 (datasheet: M01312); the renal carcinoma protocol specifies microwaving but gives no retrieval buffer (PMC4023980).
Section 2
What Is the Expected SATB1 Staining Pattern?
SATB1 should appear predominantly in nuclei, consistent with its nuclear matrix and PML body localization and lack of a transmembrane segment (UniProt Q01826). In paraffin sections, expect staining in selected neuronal and immune cell populations, especially cerebral cortex neurons and thymic cortical cells (HPA: High). HPA rates the tissue IHC pattern Enhanced, while noting medium consistency between staining and RNA expression (HPA: tissue IHC).
| Distinct nuclear staining in thymic cortical cells or cerebral cortex neurons; nearby cells may differ in intensity. | This fits the reported high staining in those cell populations (HPA: thymus cortical cells, High; cerebral cortex neuronal cells, High). Judge the relevant cells, rather than treating every nucleus in a section as an expected positive (HPA: tissue IHC profile). |
| Predominantly cytoplasmic, membranous, or extracellular staining with little nuclear signal. | That distribution is inconsistent with SATB1's nuclear matrix and PML body localization (UniProt Q01826) and the nuclear tissue pattern (HPA: tissue IHC). Treat it as suspect; review the antibody and detection controls before assigning it to SATB1 (standard IHC practice). |
| Strong staining in adipocytes or appendix glandular cells, especially without the expected nuclear pattern. | Those cell populations are reported as not detected (HPA: adipose tissue adipocytes; appendix glandular cells). Consider antibody cross-reactivity or endogenous detection activity, and compare a no-primary control; staining alone does not identify which cause applies (standard IHC practice). |
| Widespread haze or similar color over tissue and empty areas obscures cell boundaries. | This is background rather than a scorable cell-specific nuclear pattern (standard IHC practice; HPA: nuclear tissue IHC profile). Check no-primary and detection controls, blocking, washing, and chromogen development before interpreting weak nuclei (standard IHC practice). |
| No nuclear staining in thymic cortical cells or cerebral cortex neurons. | These are reported high-staining populations (HPA: thymus; cerebral cortex). An absent result warrants checking tissue identity and assay performance, including retrieval and detection controls (standard IHC practice); it does not by itself establish that the sample lacks SATB1. |
💡Expected SATB1 appearanceCall a positive result when selected cells show clear nuclear chromogen, potentially strong in thymic cortical cells or cerebral cortex neurons (HPA: High; UniProt Q01826: nuclear); diffuse cytoplasmic color or uniform staining of reported negative cells is suspect (HPA: tissue IHC; standard IHC practice).
| Cell population and tissue context | HPA reports high staining in lymph node and tonsil non-germinal center cells and placental decidual cells, alongside thymic cortical cells and cerebral cortex neurons (HPA: tissue IHC). Hippocampal neurons and ovarian follicle cells are medium (HPA: tissue IHC); a single intensity cutoff across tissues could misclassify them. |
| Nuclear distribution | UniProt places SATB1 in the nuclear matrix and, when sumoylated, PML nuclear bodies (UniProt Q01826). HPA ICC-IF reports supported nucleoplasmic and additional nuclear-body localization (HPA: subcellular). Nuclear-body signal need not make every nucleus look uniformly filled; assess the dominant nuclear pattern. |
| Antibody validation | Three listed antibodies have Enhanced IHC status: HPA051769, HPA070534, and CAB056144 (HPA: antibodies). HPA describes the tissue result as Enhanced but notes medium staining–RNA consistency (HPA: tissue IHC). Use validation as support for the reported pattern, while retaining slide controls for the antibody in use (standard IHC practice). |
| Retrieval and fixation evidence | Antigen retrieval is a routine variable in paraffin IHC (standard IHC practice). The supplied sources report no SATB1-specific fixation effect or retrieval condition (UniProt Q01826; HPA: tissue IHC). Optimize against a reported positive population and appropriate controls without inferring a target-specific sensitivity from tissue intensity. |
| What about IF/ICC? | HPA reports supported nucleoplasmic and additional nuclear-body localization in ICC-IF, with images from HEK293, RT-4, and U2OS (HPA: subcellular). This supports a localization comparison; IF/ICC assay design belongs in its separate guide and the tissue IHC result does not specify an IF protocol. |
| Situation | Likely cause | Next action |
|---|---|---|
| Expected positive tissue is blank. | The assay may have failed, or the examined field may miss the reported positive cells (HPA: thymus cortical cells, High; standard IHC practice). | Confirm the cell population, then check a known-positive section, retrieval, primary-antibody application, and detection controls (standard IHC practice). |
| Nuclei are weak but tissue shows substantial background. | Background can obscure a weak nuclear pattern; staining intensity alone cannot resolve this (standard IHC practice). | Compare no-primary controls, review blocking and washes, and assess chromogen development before scoring nuclei (standard IHC practice). |
| Most color is cytoplasmic. | The compartment conflicts with SATB1's nuclear localization (UniProt Q01826; HPA: tissue IHC). | Check the no-primary control and the antibody's IHC validation, then interpret only a reproducible nuclear pattern in expected cells (standard IHC practice; HPA: antibodies). |
| Adipocytes stain strongly. | HPA reports SATB1 as not detected in adipocytes; cross-reactivity or endogenous detection activity is possible (HPA: adipose tissue; standard IHC practice). | Inspect nuclear localization and compare no-primary and detection controls before calling the staining SATB1 positive (standard IHC practice). |
| Cerebellar granular-layer cells are negative while cortical neurons stain. | The difference matches HPA's cell-specific observations: not detected in cerebellar granular-layer cells and high in cerebral cortex neurons (HPA: tissue IHC). | Score each named population against its own reported pattern; do not use a positive cortical field to label all neuronal populations positive (HPA: tissue IHC). |
| A negative slide is being used to reject a weak positive slide. | A negative field may contain a population with low or undetected staining, such as thyroid glandular cells or appendix glandular cells (HPA: Low; Not detected). | Verify the exact tissue and cell type, and compare the weak slide with a reported high-staining population and assay controls (HPA: tissue IHC; standard IHC practice). |
Experiment evidence
IHC & IF images showing SATB1 staining
M01312 · IHCIHC analysis of STAB1 using anti-STAB1 antibody (M01312). STAB1 was detected in a paraffin-embedded section of human spleen tissue. Heat med
M01312 · IHCIHC analysis of STAB1 using anti-STAB1 antibody (M01312). STAB1 was detected in a paraffin-embedded section of human prostatic acinar adenoc
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000182568-SATB1/tissue/cerebral+cortex#imid_17130283
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000182568-SATB1/tissue/colon#imid_17130129
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000182568-SATB1/tissue/liver#imid_17130130
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000182568-SATB1/tissue/kidney#imid_17130123
HPA · ICC-IF HEK293Human Protein Atlas immunofluorescence, HEK293, antibody HPA051769; green = SATB1, blue = nucleus, red = microtubules.
HPA · ICC-IF HEK293Human Protein Atlas immunofluorescence, HEK293, antibody HPA051769; green = SATB1, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for SATB1 IHC & IF
🧪Run thymus first: cortical cells should show nuclear SATB1 staining (HPA: High in thymus cortical cells; UniProt Q01826: nuclear matrix). Use adipose tissue as the negative tissue, assessing adipocytes for absent staining (HPA: Not detected in adipocytes); on the thymus slide, unstained neighboring cells can show local background, but HPA does not identify a validated internal negative cell type there.
Positive control tissue: Cerebral cortex (Neuronal cells, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show SATB1 in HEK293, RT-4, U2OS, with annotated localisation: Nucleoplasm (supported) (HPA subcellular).
Technical controls: Include a no-primary (secondary-only) control, an isotype control matched to the actual primary antibody’s host species and clonality, and SATB1-knockout material as a biological negative (standard IHC practice). For chromogenic thymus sections, quench endogenous peroxidase and check the no-primary slide for background signal (standard IHC practice).
⚠️Feasibility: No SATB1-specific fixation window or fixation effect is reported in the supplied evidence, and SATB1 retrieval requirements remain unreported. The selected M01312 caption names STAB1 rather than SATB1 and does not report a fixative, so its EDTA retrieval conditions cannot be assigned to SATB1 (selected product caption). IF/ICC can help assess the expected nucleoplasmic and nuclear-body localization (HPA: supported subcellular locations), but the supplied evidence does not establish that frozen sections or IF are easier; assess tissue background with the controls above (standard IHC/IF practice).
HPA tissue IHC evidence for SATB1
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — Medium consistency between antibody staining and RNA expression data. External characterization data supports antibody staining.). Rows are taken directly from the HPA tissue chart — click any row's HPA link to view the source.
Positive expression · recommended positive controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Cerebral cortex | Neuronal cells | High | Protein (IHC) | HPA → |
| Lymph node | Non-germinal center cells | High | Protein (IHC) | HPA → |
| Placenta | Decidual cells | High | Protein (IHC) | HPA → |
| Thymus | Cortical cells | High | Protein (IHC) | HPA → |
| Tonsil | Non-germinal center cells | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Appendix | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Breast | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Cerebellum | Cells in granular layer | Not detected | Protein (IHC) | HPA → |
| Cervix | Glandular cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced SATB1 IHC Tips
Use nuclear localization and cell type controls to troubleshoot SATB1 staining in paraffin sections; verify the antibody identity before adopting the supplied retrieval conditions.
Which retrieval condition should I try first for weak SATB1 staining in paraffin sections?
Use heat-mediated antigen retrieval in EDTA at pH 8.0 as the first condition for this page (datasheet M01312). The supplied M01312 tissue caption names STAB1, so it does not establish SATB1 staining performance with that antibody (M01312 caption). Check the antibody identity before interpreting a weak result or carrying over its incubation and detection settings (M01312 caption). If identity is confirmed and nuclear staining remains weak, compare retrieval times on adjacent sections while keeping detection and exposure to DAB constant (standard IHC practice). Include a thymus section with cortical cells as a positive tissue control (HPA: High in thymus cortical cells).
Could fixation explain absent SATB1 nuclear staining?
Target-specific SATB1 sensitivity to fixative type or fixation duration is unknown from the supplied evidence (record; M01312 caption: fixative not stated). The M01312 caption describes a paraffin section but gives no fixative, and it names STAB1 rather than SATB1 (M01312 caption). Record the fixative and processing history for each specimen, then compare similarly processed positive and test sections in the same staining run (standard IHC practice). If nuclei are weak throughout both sections, review processing and retrieval before drawing a biological conclusion (standard IHC practice). Judge the control in cortical thymus cells, where staining is reported as High (HPA: thymus cortical cells).
What staining pattern should count as SATB1 positive?
Count a cell as SATB1 positive when convincing chromogen is confined primarily to its nucleus (UniProt Q01826: nuclear matrix; HPA: nuclear tissue expression). Nucleoplasmic staining and nuclear bodies are plausible patterns, while sumoylated SATB1 can localize to PML nuclear bodies (HPA subcellular: supported locations; UniProt Q01826: PML bodies). Interpret apparent puncta against the counterstain and section thickness, since overlapping nuclei can imitate nuclear foci (standard IHC practice). Diffuse cytoplasmic or membranous DAB alone is discordant with the supplied localization evidence (UniProt Q01826: nuclear localization and no transmembrane segment). Compare that pattern with a no-primary control before assigning positive cells (standard IHC practice).
How should isoforms and epitope location affect an unexpected staining pattern?
SATB1 has 2 listed isoforms, but the supplied evidence does not map the catalog antibody epitope to either one (UniProt Q01826: isoforms 1 and 2; supplied antibody record). Confirm the immunogen or epitope region from antibody documentation before interpreting isoform-dependent staining (standard IHC practice). The protein has a CMP region at residues 71–172 and a CUTL region at 175–248 (UniProt Q01826: domains). Acetylation at lysine 136 and phosphorylation at serine 185 are reported, but their effects on this antibody's binding are untested here (UniProt Q01826: modified residues; supplied antibody record). Test any proposed epitope explanation with an independently characterized antibody and matched sections (standard IHC practice).
How can I assess SATB1 by multiplex IF alongside a cell type marker?
For IF, pair SATB1 with a validated marker of the expected cell type and use a nuclear counterstain to assign signals to individual cells (standard IF practice). Cortical thymus cells or neuronal cells in cerebral cortex provide evidence-based contexts for selecting that marker (HPA: High in thymus cortical cells and cerebral cortex neuronal cells). Choose a fluorophore channel with low tissue autofluorescence and inspect single-color controls before interpreting overlap (standard IF practice). SATB1 is nuclear and has no transmembrane segment, so permeabilize sufficiently for antibody access to the nuclear epitope after fixation (UniProt Q01826: localization and topology; standard IF practice). Optimize fixation and permeabilization empirically because no antibody-specific IF conditions are supplied (supplied antibody record).
How do I separate nonspecific DAB background from SATB1 staining?
Compare suspect staining with a no-primary control and a matched negative cell population before changing antibody concentration (standard IHC practice). SATB1 interpretation should emphasize nuclei, whereas widespread cytoplasmic or extracellular DAB is inconsistent with its reported nuclear localization (UniProt Q01826: nuclear matrix; HPA: nuclear expression). Block endogenous peroxidase before HRP detection, and inspect whether the same deposits appear without primary antibody (standard chromogenic IHC practice). If background persists, adjust blocking, washes, or primary concentration one variable at a time while preserving the positive control (standard IHC practice). Do not adopt the M01312 caption's 1:50 setting as SATB1 validation: that caption names STAB1 (M01312 caption).
How should I quantify heterogeneous SATB1 nuclear staining across sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define positive nuclei using a fixed DAB threshold and score only the prespecified cell population (standard quantitative IHC practice). Report the percentage of positive cells and, when intensity matters, an H-score derived from the percentages at each intensity level (standard quantitative IHC practice). For spatial comparisons, report positive nuclei per mm² of evaluable tissue, excluding folds and necrotic regions (standard quantitative IHC practice). Normalize counts to the total number of eligible nuclei or sampled tissue area, as appropriate, and keep sampling rules constant across sections (standard quantitative IHC practice). Stratify by cell type because SATB1 staining differs across reported tissue cell populations (HPA: tissue IHC profile).
What findings would make a SATB1 positive call unreliable?
A credible call combines nuclear DAB with the expected cell type and a clean no-primary control (UniProt Q01826: nuclear localization; HPA: tissue IHC profile; standard IHC practice). High staining in cortical thymus cells or non-germinal center lymph node cells can support the run, but neither establishes antibody specificity alone (HPA: High in these cells; standard IHC practice). Treat isolated cytoplasmic staining, section-edge accentuation, and staining restricted to necrotic areas as potential artifacts (UniProt Q01826: nuclear localization; standard IHC practice). Check endogenous peroxidase when DAB appears in the no-primary control (standard chromogenic IHC practice). Finally, verify antibody identity: the supplied M01312 caption identifies STAB1, not SATB1 (M01312 caption).
Boster reagents
Best SATB1 / DNA-binding protein SATB1 IHC Antibodies
Both anti-SATB1 catalog entries list human, mouse, and rat reactivity (catalog: A01312-1, M01312). The supplied IHC figure caption, however, identifies STAB1 rather than SATB1 (M01312 image caption).
Real IHC data
Anti-SATB1 Rabbit Monoclonal Antibody
Cat # M01312
M01312 is the sole rendered card; its IHC caption describes staining in a paraffin-embedded human spleen section at 1:50, but identifies the target as STAB1 (M01312 image caption). A01312-1 lists frozen-section IHC and ICC without a supplied image, while M01312 lists IHC and ICC/IF without a supplied IF image (catalog: A01312-1, M01312).
Which to pick: For tissue IHC, M01312 is the pictured rabbit monoclonal option, but confirm target identity before treating its STAB1-captioned paraffin-section image as SATB1 evidence; the fixative is unreported (catalog: M01312; M01312 image caption). For IF/ICC, M01312 explicitly lists IF and ICC, while A01312-1 lists ICC; neither has a supplied IF image (catalog: M01312, A01312-1). Both list human, mouse, and rat reactivity, but the supplied tissue caption documents only a human section (catalog: A01312-1, M01312; M01312 image caption).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q01826 (SATB1_HUMAN, DNA-binding protein SATB1).
- Human Protein Atlas. SATB1 tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. SATB1 subcellular location (ICC-IF): Mainly localized to the nucleoplasm. In addition localized to the nuclear bodies..
- Human Protein Atlas. SATB1 antibody validation summary (3 antibodies).
- Expression of the global regulator SATB1 is an independent factor of poor prognosis in high grade epithelial ovarian cancer. Journal of ovarian research 2012 — PMC3472180.
- Overexpression of SATB1 is associated with biologic behavior in human renal cell carcinoma. PloS one 2014 — PMC4023980.
- Special AT-rich DNA-binding protein-1 expression is associated with liver cancer metastasis. Oncology letters 2016 — PMC5228311.
- Molecular correlates and prognostic significance of SATB1 expression in colorectal cancer. Diagnostic pathology 2012 — PMC3523011.
- PubMed PMID:1505028 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
- PubMed PMID:16641997 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for SATB1 (DNA-binding protein SATB1). Expected localisation Nuclear staining (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet M01312). Validated IHC antibody: M01312 at 1:50 (the product's own IHC validation image is shown). Controls (HPA): positive Cerebral cortex (Neuronal cells); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human spleen tissue; fixative not specified (datasheet M01312); 2. Fixation: Image fixative and duration unreported (datasheet M01312); verify before use.; 3. Sectioning: 4–5 µm sections on charged slides (standard); 4. Deparaffinisation: Xylene, graded ethanol series to water (standard); 5. Antigen retrieval: Heat retrieval: EDTA pH 8.0 (datasheet M01312); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet M01312); 8. Primary antibody: Rabbit monoclonal (clone ACO-19) anti-SATB1, 1:50 (datasheet M01312); 9. Primary incubation: Overnight at 4 °C (datasheet M01312). Watch for: Fixation: Keep fixation consistent across sections (standard IHC practice; not target-specific); Caveat: Lymphoid staining is strongest in non-germinal center cells (HPA tissue IHC); Isoform / epitope: Two isoforms; verify antibody epitope coverage (UniProt); Regulation: Predominantly expressed in thymus (UniProt). Evidence: UniProt Q01826, HPA tissue IHC · Enhanced, HPA ICC-IF · Mainly localized to the nucleoplasm. In addition localized to the…, Datasheet M01312, PMC3472180, PMC4023980. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/SATB1/immunohistochemistry-ihc-guide.