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SWAP70 / Switch-associated protein 70 · IHC design guide
Design Immunohistochemistry for SWAP70
Plan chromogenic SWAP70 IHC on paraffin sections with the catalog antibody at 2–5 μg/ml (datasheet A06972-3). Use spleen white pulp as a positive reference and score cytoplasmic staining by cell population (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
SWAP70 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 | Cytoplasmic in a subset of immune cells (HPA tissue IHC) | |
| Staining pattern | High cytoplasmic staining in some immune cells (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A06972-3) | |
| Positive control | Appendix+4 more · see all | |
| Negative control | Adipose tissue+4 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A06972-3) | |
| Caveat | Immune-cell proportions can change section-level signal (HPA tissue IHC) | |
| Regulation | B-cell activation shifts SWAP70 location (UniProt) | |
| Isoform / epitope | No listed isoforms; one chain and no extracellular domain (UniProt) |
Section 1
Recommended SWAP70 IHC & IF Protocols
Compare the catalog antibody’s IHC-P protocol with the published SWAP70 staining protocol for lower-grade glioma tissue (PMC9885112).
| Sample | Paraffin-embedded human ovary serous carcinoma tissue; fixative not specified (datasheet A06972-3) |
| Fixation | Image fixative and duration unreported (datasheet A06972-3); 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 A06972-3); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A06972-3) |
| Primary antibody | Rabbit anti-SWAP70, 2-5 μg/ml (datasheet A06972-3) |
| Primary incubation | Overnight at 4 °C (datasheet A06972-3) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A06972-3) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | SWAP70-positive staining in lymphoid tissue of appendix (HPA tissue IHC: High). HPA tissue profile: High cytoplasmic expression in a subset of immune cells. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA retrieval at pH 8.0 (datasheet A06972-3); the published protocol does not specify retrieval conditions (PMC9885112).
Section 2
What Is the Expected SWAP70 Staining Pattern?
SWAP70 staining should be assessed by cell type and compartment: HPA reports high cytoplasmic staining in a subset of immune cells, including germinal center and splenic white pulp cells (HPA: tissue IHC). SWAP70 has no transmembrane segment and can localize to cytoplasm, membrane and nucleus depending on context (UniProt Q9UH65: topology and localization). HPA rates the tissue IHC pattern Supported, with medium consistency between staining and RNA expression (HPA: tissue IHC reliability).
| Strong cytoplasmic staining in tonsil or lymph node germinal center cells, or splenic white pulp cells. | This matches reported high staining in those compartments (HPA: tissue IHC). Score the relevant cells rather than treating every cell in the section as an expected positive; HPA describes high cytoplasmic expression in a subset of immune cells (HPA: tissue IHC). |
| Predominantly nuclear or uniformly membranous staining across the section, with little cytoplasmic signal in expected positive cells. | Check for a staining artefact before accepting this as the IHC pattern (HPA: high cytoplasmic expression in a subset of immune cells; general IHC practice). Nuclear or membrane localization is possible in particular B-cell activation states, so compartment alone cannot settle specificity (UniProt Q9UH65: localization). |
| Strong staining of bronchial respiratory epithelial cells or adipocytes, especially when expected immune cells do not stain. | Those cell types are reported as not detected (HPA: bronchus respiratory epithelium; adipose adipocytes). Investigate antibody cross-reactivity or endogenous chromogenic detection activity (general IHC practice). Staining of nearby immune cells requires separate cell-level assessment (HPA: subset of immune cells). |
| Diffuse chromogen over cells and extracellular areas, without clear cell boundaries or enrichment in expected positive cells. | Treat this as background until controlled: inadequate blocking, detection reagent background or excessive primary antibody can produce diffuse staining (general IHC practice). Compare a no-primary control and a known-positive section processed in the same run (general IHC practice). |
| No signal in tonsil germinal center cells or splenic white pulp cells. | These are reported high-staining populations, so an absent result calls for a run-level check before biological interpretation (HPA: tonsil and spleen tissue IHC; general IHC practice). Review section identity, antibody preparation, retrieval and detection controls (general IHC practice). |
💡Expected SWAP70 appearanceCall an IHC result positive when identifiable tonsil or lymph node germinal center cells, or splenic white pulp cells, show high cytoplasmic staining (HPA: tissue IHC); broad staining of adipocytes or bronchial respiratory epithelial cells is suspect because those cells are reported as not detected (HPA: tissue IHC).
| Cell population and tissue context | HPA reports high staining in germinal center cells, splenic white pulp cells, appendix lymphoid tissue and placental cytotrophoblasts; bone marrow hematopoietic cells are medium, while lung macrophages are low (HPA: tissue IHC). Choose and score the named cell population within each tissue (general IHC practice). |
| Intracellular distribution | Cytoplasmic staining is the main tissue IHC pattern (HPA: tissue IHC). Resting B-cells are mainly cytoplasmic; activation can recruit SWAP70 to the plasma membrane and then nucleus (UniProt Q9UH65: localization). Interpret a changed compartment alongside cell identity and activation context (general IHC practice). |
| Antibody evidence | HPA006810 has Supported IHC status, while HPA006929 has Supported ICC status (HPA: antibody validation). HPA rates the tissue pattern Supported but notes medium consistency with RNA expression and staining in some unannotated cells or structures (HPA: tissue IHC reliability). Resolve uncertain cells by morphology and controls (general IHC practice). |
| Topology and processing | SWAP70 has no transmembrane segment, signal peptide or propeptide, and is listed as one 1–585 chain (UniProt Q9UH65: topology and processing). Membrane-associated signal should be interpreted as localization of the protein, without assuming a membrane-spanning or shed form (UniProt Q9UH65: topology and localization). |
| Target-specific retrieval sensitivity | No SWAP70-specific fixation or antigen-retrieval effect is established by the supplied HPA tissue IHC and UniProt records (HPA: tissue IHC scope; UniProt Q9UH65: record scope). Assess retrieval and detection with run controls as general IHC practice; do not infer a SWAP70-specific effect from tissue staining levels. |
| Situation | Likely cause | Next action |
|---|---|---|
| Expected germinal center or splenic white pulp cells are unstained. | A section, primary antibody, retrieval or detection problem is possible (general IHC practice); both populations are reported high by IHC (HPA: tonsil and spleen). | Confirm the intended cell population on the section, then check reagent preparation, retrieval conditions and detection controls against a known-positive section in the same run (general IHC practice). |
| The entire section has weak, diffuse brown signal. | Background from the detection system, incomplete blocking or excessive antibody can obscure cell-specific staining (general IHC practice). | Compare the no-primary control; review blocking, primary antibody concentration, washes and chromogen development, then rescore only distinguishable cells (general IHC practice). |
| Bronchial respiratory epithelium or adipocytes stain strongly. | These cells are reported as not detected (HPA: bronchus and adipose tissue IHC). Cross-reactivity or endogenous detection activity is possible (general IHC practice). | Confirm cell identity, inspect a no-primary control and compare staining with an expected positive tissue before assigning SWAP70 positivity (general IHC practice; HPA: tonsil and spleen tissue IHC). |
| Signal appears mainly nuclear or at the cell edge. | SWAP70 can redistribute during B-cell activation, although HPA describes the tissue IHC pattern as mainly cytoplasmic in a subset of immune cells (UniProt Q9UH65: localization; HPA: tissue IHC). Artefact is also possible (general IHC practice). | Identify the stained cells and check whether expected cytoplasmic positives are present; compare controls before interpreting compartment shifts (general IHC practice). |
| Only a few cells stain in a tissue otherwise called negative. | HPA describes expression in a subset of immune cells, and its listed negatives refer to specified cell types, such as adipocytes or bronchial respiratory epithelial cells (HPA: tissue IHC). | Score stained cells by morphology and location; report the named negative cell population separately from any infiltrating immune cells (general IHC practice; HPA: tissue IHC). |
| IF/ICC: where should SWAP70 appear? | HPA reports plasma membrane as the main supported ICC-IF location, with actin filaments as an additional supported location (HPA: subcellular ICC-IF). This evidence concerns ICC-IF images, not the tissue IHC pattern (HPA: subcellular ICC-IF and tissue IHC). | Interpret membrane and actin-associated IF signal in its own cell context; use the separate IF/ICC guide for the assay workflow (HPA: subcellular ICC-IF; general assay practice). |
Experiment evidence
IHC & IF images showing SWAP70 staining
A06972-3 · IHCIHC analysis of SWAP70 using anti-SWAP70 antibody (A06972-3). SWAP70 was detected in a paraffin-embedded section of human ovary serous carci
A06972-3 · IHCIHC analysis of SWAP70 using anti-SWAP70 antibody (A06972-3). SWAP70 was detected in a paraffin-embedded section of human rectum adenocarcin
A06972-3 · IFIF analysis of SWAP70 using anti-SWAP70 antibody (A06972-3). SWAP70 was detected in a paraffin-embedded section of human colon cancer tissue
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000133789-SWAP70/tissue/cerebral+cortex#imid_2105400
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000133789-SWAP70/tissue/colon#imid_2089702
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000133789-SWAP70/tissue/liver#imid_2089719
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000133789-SWAP70/tissue/kidney#imid_2089704
HPA · ICC-IF HUVEC/TERT2Human Protein Atlas immunofluorescence, HUVEC/TERT2, antibody HPA006929; green = SWAP70, blue = nucleus, red = microtubules.
HPA · ICC-IF HUVEC/TERT2Human Protein Atlas immunofluorescence, HUVEC/TERT2, antibody HPA006929; green = SWAP70, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for SWAP70 IHC & IF
🧪Run tonsil first and look for staining in germinal center cells (HPA: High in tonsil germinal center cells). Use adipose tissue as the negative, where adipocytes should be unstained (HPA: Not detected in adipose tissue adipocytes); on the tonsil slide, use cells outside germinal centers as an internal background reference and verify that they show no specific staining, since their expression is not specified in the supplied HPA row.
Positive control tissue: Appendix (Lymphoid tissue, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show SWAP70 in HUVEC/TERT2, RT-4, U2OS, with annotated localisation: Plasma membrane (supported) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only slide and a matched nonimmune rabbit IgG isotype control (catalog caption: rabbit primary antibody), plus SWAP70 knockout material as a biological negative. Block endogenous peroxidase and check for background DAB signal in tonsil before scoring (catalog caption: peroxidase detection with DAB).
⚠️Feasibility: The selected A06972-3 paraffin-section caption does not report a fixative, and no target-specific fixation window or fixation effect is reported in the supplied evidence (catalog caption: fixative not stated). Heat retrieval in EDTA at pH 8.0 was used for the captioned ovary carcinoma section; its necessity for tonsil is unreported (catalog caption: heat-mediated EDTA retrieval). Frozen-section IHC is not documented here, and the supplied ICC-IF localization images do not establish that tissue IF is easier (HPA: ICC-IF images in HUVEC/TERT2, RT-4 and U2OS); assess tonsil DAB background with the control slides (catalog caption: DAB detection).
HPA tissue IHC evidence for SWAP70
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Supported — Medium consistency between antibody staining and RNA expression data. Antibody staining in cells/structures not annotated, view images.). 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 |
|---|---|---|---|---|
| Appendix | Lymphoid tissue | High | Protein (IHC) | HPA → |
| Lymph node | Germinal center cells | High | Protein (IHC) | HPA → |
| Placenta | Cytotrophoblasts | High | Protein (IHC) | HPA → |
| Spleen | Cells in white pulp | High | Protein (IHC) | HPA → |
| Tonsil | 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 → |
| Adrenal gland | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Breast | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | Not detected | Protein (IHC) | HPA → |
| Caudate | Glial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced SWAP70 IHC Tips
Troubleshoot SWAP70 chromogenic IHC in paraffin sections using the catalog antibody’s tissue evidence and compartment specific controls.
Which antigen retrieval should I start with for SWAP70 paraffin sections?
Begin with heat mediated retrieval in EDTA at pH 8.0 (datasheet A06972-3). The catalog antibody produced SWAP70 staining in a paraffin section after this retrieval, followed by 2 μg/ml primary antibody overnight at 4°C (IHC caption A06972-3). If staining is weak, check that sections were fully deparaffinized and compare a known positive germinal center region processed in the same run (HPA: High in tonsil germinal center cells). Keep retrieval conditions identical across comparison sections, because differences in heating can change apparent staining intensity (standard IHC practice). Record the heating time and temperature before adjusting antibody concentration (standard IHC practice).
How should I assess whether fixation caused weak SWAP70 staining?
SWAP70 specific sensitivity to fixation is unknown from the supplied evidence; the catalog image identifies a paraffin section but does not state its fixative (IHC caption A06972-3). Record the fixative and fixation duration for each specimen, then compare affected sections with a positive control processed under matching conditions (standard IHC practice). Use the documented EDTA pH 8.0 retrieval and 2 μg/ml primary antibody conditions as the initial comparison settings (IHC caption A06972-3). If staining varies with fixation history, test paired sections while holding retrieval and detection constant (standard IHC practice). Do not infer a SWAP70 specific fixation effect from tissue staining patterns alone.
What cellular pattern should I expect in SWAP70 IHC?
Expect prominent cytoplasmic staining in a subset of immune cells in tissue sections (HPA: High cytoplasmic expression in a subset of immune cells). Germinal center cells in tonsil and lymph node offer high staining reference populations (HPA: High in tonsil and lymph node germinal center cells). SWAP70 can also occupy the plasma membrane, nucleus, and actin associated projections, with redistribution described after B cell activation (UniProt Q9UH65 subcellular location). HPA cell imaging supports plasma membrane and actin filament localization, so assess membrane accentuation in cellular context (HPA: supported subcellular locations). Compare compartments within intact cells before interpreting diffuse extracellular chromogen as target staining (standard IHC practice).
Could epitope placement explain an unexpected SWAP70 staining pattern?
Check the catalog antibody’s stated immunogen or epitope before attributing a staining difference to SWAP70 sequence features; the supplied tissue caption does not identify that epitope (IHC caption A06972-3). The reference record lists a 585 residue chain, a PH domain at residues 210–306, and no alternative isoforms (UniProt Q9UH65 record). It lists no transmembrane segment, signal peptide, glycosylation sites, or modified residues (UniProt Q9UH65 record). Those annotations cannot establish whether fixation masks this antibody’s epitope, so compare retrieval conditions on matched sections if signal is weak (standard IHC practice). Treat unexpected compartment staining as a validation question rather than evidence of an unlisted isoform.
How can IF help evaluate a disputed SWAP70 IHC pattern?
On a matched section, multiplex SWAP70 with a validated marker for the expected immune cell population to check whether the IHC positive cells are correctly identified (HPA: High cytoplasmic expression in a subset of immune cells). Choose fluorophores in red or far red channels after inspecting unstained tissue for autofluorescence, and include single stain controls (standard IF practice). For an intracellular epitope, use controlled permeabilisation because SWAP70 lacks a transmembrane segment and has cytoplasmic and membrane associated locations (UniProt Q9UH65 topology and subcellular location). Optimize permeabilisation against morphology and background; the catalog’s paraffin IHC caption supplies no IF permeabilisation condition (IHC caption A06972-3). Interpret IF as a compartment check alongside the chromogenic result.
What should I check when SWAP70 DAB staining is diffuse?
Run a section without primary antibody to reveal detection reagent or endogenous peroxidase signal, and apply a peroxidase block as part of the chromogenic workflow (standard IHC practice). The documented assay used a peroxidase conjugated secondary and DAB, with 10% goat serum blocking and 2 μg/ml primary antibody (IHC caption A06972-3). If background persists, inspect reagent only controls and titrate primary antibody while keeping development time consistent (standard IHC practice). Compare staining with a positive germinal center region and a low signal cell population such as adipocytes (HPA: High in tonsil germinal center cells; Not detected in adipose adipocytes). Exclude folds, section edges, and necrotic areas from assessment (standard IHC practice).
How should I quantify SWAP70 staining across sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring, because tissue staining is concentrated in subsets of immune cells (HPA: High cytoplasmic expression in a subset of immune cells). For chromogenic IHC, report the percentage of positive cells and an intensity based H-score within annotated regions, or positive cell density per mm² when cell numbers differ (standard IHC practice). Normalize positive cell counts to the number of eligible cells or the analyzed tissue area, and report those denominators (standard IHC practice). Keep retrieval, primary incubation, DAB development, counterstain, and image analysis thresholds constant across the comparison (standard IHC practice). Score cytoplasmic and membrane associated signal separately if the study tests redistribution (UniProt Q9UH65 subcellular location).
How do I distinguish genuine SWAP70 signal from artefact?
A credible positive signal should occur in intact cells and align with a plausible SWAP70 compartment, especially cytoplasm in immune cell subsets (HPA: High cytoplasmic expression in a subset of immune cells). Compare suspect staining with germinal center cells in tonsil or lymph node, which have high reported expression (HPA: High in tonsil and lymph node germinal center cells). Membrane or nuclear signal can be biologically plausible after B cell activation, but requires cellular context and suitable controls (UniProt Q9UH65 subcellular location). Discount chromogen concentrated at section edges, folds, or necrotic areas, and investigate staining that persists without primary antibody as potential endogenous enzyme or detection background (standard IHC practice). A wrong cell population warrants further validation.
Boster reagents
Best SWAP70 / Switch-associated protein 70 IHC Antibodies
Anti-SWAP70 antibody A06972-3 has IHC and IF image data from human paraffin-embedded cancer sections (catalog: A06972-3 image captions); the fixative is unreported (catalog: A06972-3 image captions).
Real IHC data
Anti-SWAP70 Antibody ®
Cat # A06972-3
A06972-3 has IHC images from human ovary serous carcinoma and rectum adenocarcinoma paraffin sections (catalog: A06972-3 IHC image captions). The same SKU has IF images from human colon and ovarian cancer paraffin sections (catalog: A06972-3 IF image captions).
Which to pick: For tissue IHC, choose A06972-3: its IHC caption documents staining of a human paraffin section at 2 μg/ml; the fixative is unreported (catalog: A06972-3 IHC image caption). For IF, A06972-3 has human paraffin-section images at 5 μg/ml; ICC validation and clonality are unreported (catalog: A06972-3 applications, IF image captions, clone field). No cross-species choice is supported because A06972-3 lists human reactivity only (catalog: A06972-3 reactivity).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q9UH65 (SWP70_HUMAN, Switch-associated protein 70).
- Human Protein Atlas. SWAP70 tissue IHC expression (reliability: Supported).
- Human Protein Atlas. SWAP70 subcellular location (ICC-IF): Mainly localized to the plasma membrane. In addition localized to the actin filaments..
- Human Protein Atlas. SWAP70 antibody validation summary (2 antibodies).
- Integrated analysis of genome-wide DNA methylation and cancer-associated fibroblasts identified prognostic biomarkers and immune checkpoint blockade in lower grade gliomas. Frontiers in oncology 2022 — PMC9885112.
- PubMed PMID:10681448 — UniProt-cited evidence.
- PubMed PMID:11726218 — UniProt-cited evidence.
- PubMed PMID:9734811 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for SWAP70 (Switch-associated protein 70). Expected localisation Cytoplasmic in a subset of immune cells (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A06972-3). Validated IHC antibody: A06972-3 at 2-5 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Appendix (Lymphoid tissue); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human ovary serous carcinoma tissue; fixative not specified (datasheet A06972-3); 2. Fixation: Image fixative and duration unreported (datasheet A06972-3); 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 A06972-3); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A06972-3); 8. Primary antibody: Rabbit anti-SWAP70, 2-5 μg/ml (datasheet A06972-3); 9. Primary incubation: Overnight at 4 °C (datasheet A06972-3). Watch for: Fixation: Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A06972-3); Caveat: Immune-cell proportions can change section-level signal (HPA tissue IHC); Isoform / epitope: No listed isoforms; one chain and no extracellular domain (UniProt); Regulation: B-cell activation shifts SWAP70 location (UniProt). Evidence: UniProt Q9UH65, HPA tissue IHC · Supported, HPA ICC-IF · Mainly localized to the plasma membrane. In addition localized to…, Datasheet A06972-3, PMC9885112. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/SWAP70/immunohistochemistry-ihc-guide.