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TPSAB1 / Tryptase alpha/beta-1 · IHC design guide
Design Immunohistochemistry for TPSAB1
Plan chromogenic TPSAB1 IHC in paraffin sections with the catalog antibody at 0.5–1 μg/ml (datasheet: PB10016). Expect cytoplasmic staining in leukocyte subsets, probably mast cells (HPA tissue IHC), and account for possible detection of protein from more than one gene (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
TPSAB1 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 staining in leukocyte subsets (HPA tissue IHC) | |
| Staining pattern | Subset of leukocytes, probably mast cells: cytoplasmic (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet PB10016) | |
| Positive control | Bone marrow+2 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 | Antibody may detect protein from more than one gene (HPA tissue IHC) | |
| Regulation | Degranulation releases tryptase (UniProt) | |
| Isoform / epitope | Two isoforms; mature chain starts at residue 31 (UniProt) |
Section 1
Recommended TPSAB1 IHC & IF Protocols
The catalog antibody’s IHC-P protocol is presented alongside published chromogenic TPSAB1 protocols for mouse ovary and rat synovium (PMC3790711; PMC13460010).
| Sample | Paraffin-embedded human lung cancer tissue; fixative not specified (datasheet PB10016) |
| Fixation | Image fixative and duration unreported (datasheet PB10016); 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 PB10016); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet PB10016) |
| Primary antibody | Rabbit anti-TPSAB1, 0.5-1μg/ml (datasheet PB10016) |
| Primary incubation | Overnight at 4 °C (datasheet PB10016) |
| Detection | Streptavidin-biotin complex (SABC), DAB chromogen (datasheet PB10016) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | TPSAB1-positive staining in hematopoietic cells of bone marrow (HPA tissue IHC: Medium). HPA tissue profile: Selective cytoplasmic expression in subsets of leukocytes, probably mast cells. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA, pH 8.0 (datasheet PB10016); citrate, pH 6.0, is a published alternative for mouse ovary (PMC3790711).
Section 2
What Is the Expected TPSAB1 Staining Pattern?
TPSAB1 is a secreted protease released from mast-cell granules and has no transmembrane segment (UniProt Q15661). In tissue IHC, expect selective cytoplasmic staining in subsets of leukocytes, probably mast cells (HPA: tissue IHC profile). HPA rates the tissue pattern Supported, while reporting very low agreement with RNA expression and warning that the antibodies may recognize proteins from more than one gene (HPA: tissue IHC reliability).
| Discrete cytoplasmic staining in scattered leukocytes, with surrounding cells largely unstained. | This matches the selective pattern HPA describes in leukocyte subsets, probably mast cells (HPA: tissue IHC profile). Granule-associated signal is plausible because tryptase is released from mast-cell secretory granules (UniProt Q15661). The pattern supports interpretation but does not identify every positive cell as a mast cell (HPA: tissue IHC profile). |
| Strong, uniform nuclear or membrane outlining signal replaces the selective cytoplasmic pattern. | Treat this as a pattern mismatch and check the assay before scoring it as TPSAB1. HPA reports cytoplasmic staining, while UniProt describes a secreted protein without a transmembrane segment (HPA: tissue IHC profile; UniProt Q15661). Review morphology and controls for staining artifact (general IHC practice). |
| Broad staining appears in adipocytes, respiratory epithelium, or other unexpected cells. | HPA reports no detection in adipocytes of adipose tissue or breast, or in bronchial respiratory epithelial cells (HPA: tissue IHC). Consider antibody cross-reactivity or endogenous chromogenic activity, and compare a no-primary control (HPA: multigene-target caution; general IHC practice). Do not apply those negatives to every cell in those tissues. |
| Diffuse color covers stroma, lumens, and many cell types without clear cell borders. | Background or extracellular protein can obscure selective cellular staining (general IHC practice; UniProt Q15661: secreted protein). Because secretion can separate protein location from its RNA source, do not assign diffuse staining to a cell type solely from nearby RNA expression (HPA: tissue IHC reliability). |
| No convincing staining appears in a bone marrow, lymph-node, or tonsil section. | First confirm the relevant cell population is present: HPA reports Medium staining in bone-marrow hematopoietic cells and non-germinal-center cells of lymph node and tonsil (HPA: tissue IHC). A negative run calls for control and assay review; HPA's reported levels do not guarantee every section will contain positive cells. |
💡Expected TPSAB1 appearanceCall the IHC result convincing when selective cytoplasmic signal marks scattered leukocytes, probably mast cells, with roughly Medium staining in HPA-reported positive cell populations; broad nuclear, membrane, or unrelated-cell staining is a pattern mismatch (HPA: tissue IHC profile and levels; UniProt Q15661: secreted, no transmembrane segment).
| Cell identity and tissue context | HPA's positive calls concern specific populations: bone-marrow hematopoietic cells and non-germinal-center cells in lymph node and tonsil, each at Medium level (HPA: tissue IHC). Its negative calls likewise name cell populations, such as adipocytes and bronchial respiratory epithelial cells (HPA: tissue IHC). Score cells, not whole organs. |
| Secretion and processing | UniProt lists a signal peptide at residues 1–18, a propeptide at 19–30, and the mature chain at 31–275; tryptase is released on mast-cell activation (UniProt Q15661). Extracellular signal therefore needs morphological context. The supplied record does not locate the catalog antibody's epitope, so processing cannot predict its staining strength. |
| Isoforms and endothelial staining | UniProt reports two isoforms and describes isoform 2 in endothelial cells of some vessels around the aorta and breast tumor, with little if any mast-cell protein there (UniProt Q15661). Do not automatically label every endothelial signal an artifact. HPA separately reports no detection in cerebral-cortex endothelial cells (HPA: tissue IHC); site matters. |
| Antibody specificity and evidence | Four listed HPA antibodies have Supported IHC status, but HPA warns that staining may reflect proteins from more than one gene and shows very low agreement with RNA expression (HPA: antibody validation; tissue IHC reliability). Interpret a matching pattern as evidence of tryptase-associated staining, not proof that every signal is uniquely TPSAB1. |
| IF/ICC: what should a positive cell show? | HPA reports vesicular localization in ICC-IF, with the main location approved as vesicles (HPA: subcellular). This is consistent with UniProt's secretory-granule description (UniProt Q15661). Use the separate IF/ICC guide for that application; these IHC tissue levels do not set an IF intensity threshold. |
| Situation | Likely cause | Next action |
|---|---|---|
| The reported positive tissue is blank. | The sampled section may lack the relevant cells, or the staining run may have failed; HPA's Medium calls apply to named cell populations (HPA: tissue IHC; general IHC practice). | Check tissue morphology and a positive-run control, then review antibody dilution, retrieval, detection, and counterstain settings (general IHC practice). No TPSAB1-specific retrieval or fixation sensitivity is established by the supplied sources. |
| The entire section develops color. | Excess background or endogenous chromogenic activity may overwhelm selective staining (general IHC practice). HPA describes selective cytoplasmic leukocyte staining (HPA: tissue IHC profile). | Compare a no-primary control; review blocking, washes, detection exposure, and counterstain (general IHC practice). Assess whether discrete cytoplasmic positive cells remain visible. |
| Adipocytes or bronchial respiratory epithelial cells stain prominently. | Those named cell types were not detected in HPA tissue IHC; antibody cross-reactivity or detection background is possible (HPA: tissue IHC and multigene-target caution; general IHC practice). | Review cell identity and no-primary staining, then compare the suspect pattern with a Supported IHC antibody where available (HPA: antibody validation; general IHC practice). |
| Nuclear or continuous membrane staining dominates. | The compartment conflicts with HPA's cytoplasmic pattern and UniProt's secreted, non-transmembrane annotation (HPA: tissue IHC profile; UniProt Q15661). | Check morphology, no-primary background, and detection settings before assigning a positive score (general IHC practice). |
| Vascular endothelial cells stain. | Some endothelial staining may reflect isoform 2 near the aorta or breast tumor, although HPA reports no detection in cerebral-cortex endothelial cells (UniProt Q15661; HPA: tissue IHC). | Record the vessel site and cell identity. Compare with the expected local pattern and controls; avoid a blanket positive or artifact call across tissues (UniProt Q15661; HPA: tissue IHC; general IHC practice). |
| Protein staining and local RNA expression disagree. | HPA reports very low staining–RNA consistency and notes that secreted protein can lie away from the cells producing it (HPA: tissue IHC reliability). | Score the observed cellular and extracellular pattern with tissue morphology and staining controls; report the RNA discrepancy as a limitation rather than using RNA alone to override the slide (HPA: tissue IHC reliability; general IHC practice). |
Experiment evidence
IHC & IF images showing TPSAB1 staining
PB10016 · IHCIHC analysis of Mast Cell Tryptase using anti-Mast Cell Tryptase antibody (PB10016). Mast Cell Tryptase was detected in a paraffin-embedded
PB10016 · IHCIHC analysis of Mast Cell Tryptase using anti-Mast Cell Tryptase antibody (PB10016). Mast Cell Tryptase was detected in a paraffin-embedded
A08039-1 · IHCImmunohistochemical analysis of paraffin-embedded human tonsil tissue using anti-Mast Cell Tryptase antibody. Counter stained with hematoxyl
M08039 · IHCImmunohistochemical analysis of paraffin-embedded human kidney, using Mast Cell Tryptase Antibody.
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000172236-TPSAB1/tissue/cerebral+cortex#imid_6119712
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000172236-TPSAB1/tissue/colon#imid_6119654
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000172236-TPSAB1/tissue/liver#imid_6119647
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000172236-TPSAB1/tissue/kidney#imid_6119640
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody CAB016369; green = TPSAB1, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody CAB016369; green = TPSAB1, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for TPSAB1 IHC & IF
🧪Run bone marrow first; hematopoietic cells should stain (HPA: Medium in bone marrow hematopoietic cells). Use adipose tissue as the negative comparator, where adipocytes should lack staining (HPA: Not detected in adipocytes); on the bone marrow slide, assess morphologically distinct unstained cells for background.
Positive control tissue: Bone marrow (Hematopoietic cells, HPA Medium)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show TPSAB1 in A-431, U-251MG, U2OS, with annotated localisation: Vesicles (approved) (HPA subcellular).
Technical controls: Include a no-primary (secondary-only) slide, a concentration-matched rabbit IgG isotype control (selected caption: rabbit primary antibody), and a TPSAB1 knockout specimen as a biological negative. Check endogenous peroxidase activity in bone marrow and endogenous biotin background if using the caption’s avidin–biotin detection method (selected caption: SABC with DAB).
⚠️Feasibility: A target-specific fixation window and fixation effect are unreported, and the selected PB10016 paraffin-section caption does not state the fixative. The caption reports heat retrieval in EDTA at pH 8.0, but does not establish whether retrieval is required for every specimen (selected caption: EDTA retrieval). The supplied evidence does not establish that frozen sections or IF are easier; bone marrow peroxidase activity can complicate chromogenic interpretation (standard IHC practice).
HPA tissue IHC evidence for TPSAB1
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Supported — Very low consistency between antibody staining and RNA expression data. Caution, targets protein from more than one gene. Secreted protein, tissue location of RNA and protein is expected to differ. 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 |
|---|---|---|---|---|
| Bone marrow | Hematopoietic cells | Medium | Protein (IHC) | HPA → |
| Lymph node | Non-germinal center cells | Medium | Protein (IHC) | HPA → |
| Tonsil | Non-germinal center cells | Medium | 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 → |
| Appendix | Endocrine cells | Not detected | Protein (IHC) | HPA → |
| Breast | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced TPSAB1 IHC Tips
Troubleshoot TPSAB1 staining by assessing retrieval, tissue processing, expected localisation and cell identity before comparing signal intensity across sections.
What retrieval conditions should I try when TPSAB1 staining is weak?
Start with heat-mediated retrieval in EDTA at pH 8.0 for paraffin sections (datasheet PB10016). The selected image used this retrieval before overnight incubation at 4°C with 1 μg/ml catalog antibody, so assess retrieval alongside those documented conditions (datasheet PB10016). If staining remains weak, vary heating time on matched sections while keeping detection and development constant (standard IHC practice). Include a mast-cell-containing control section and inspect tissue integrity: excessive heating can damage morphology, while inadequate retrieval can leave epitopes inaccessible (standard IHC practice). Record the heating method and duration because the caption supplies neither (datasheet PB10016).
Could fixation explain weak or uneven TPSAB1 staining?
Target-specific fixation sensitivity is unknown because the selected paraffin-section caption does not state a fixative (datasheet PB10016). Record each specimen’s fixative and fixation duration, then compare sections processed together under the documented EDTA pH 8.0 retrieval conditions (datasheet PB10016; standard IHC practice). Check whether uneven staining follows differences in tissue thickness, processing or retrieval exposure before changing antibody concentration (standard IHC practice). Use a consistently processed control section to distinguish a preparation problem from a staining-run problem (standard IHC practice). Neither the tissue staining profile nor TPSAB1’s molecular features establishes a fixation preference (HPA tissue IHC; UniProt Q15661).
Where should convincing TPSAB1 signal appear in a paraffin section?
Look for cytoplasmic staining in a subset of leukocytes consistent with mast cells (HPA tissue IHC: selective cytoplasmic expression, probably mast cells). A punctate or granular pattern is biologically plausible because TPSAB1 is stored in secretory granules and released during activation (UniProt Q15661: subcellular location and function). Evaluate extracellular staining cautiously: release is possible, but diffuse deposit alone cannot identify its cellular source (UniProt Q15661: secreted; standard IHC interpretation). TPSAB1 has no transmembrane segment, so a crisp membrane-only pattern needs independent confirmation (UniProt Q15661: topology). Compare morphology and nearby cells before assigning the signal to mast cells (standard IHC practice).
Can this stain distinguish TPSAB1 isoforms or identify the bound epitope?
Do not assign an isoform from staining intensity alone: the record lists 2 TPSAB1 isoforms, but the selected caption does not identify the antibody’s epitope (UniProt Q15661: isoforms; datasheet PB10016). Signal peptide residues 1–18 and propeptide residues 19–30 are processed before the mature chain spanning 31–275 (UniProt Q15661: processing). Glycosylation sites at 132 and 233 could matter to epitope accessibility only if the binding site is known (UniProt Q15661: glycosylation; standard IHC interpretation). Obtain epitope or isoform-specific validation before interpreting distinct cell patterns as isoform differences (standard IHC practice).
How should I check an IF multiplex result against the chromogenic IHC pattern?
Use the chromogenic section to identify candidate cytoplasmic, granular TPSAB1-positive cells, then test cell identity in a separate IF multiplex with a validated mast-cell marker (HPA tissue IHC: probable mast cells; UniProt Q15661: secretory granules). Choose fluorophores after measuring tissue autofluorescence and include single-stain controls to assess channel bleed-through (standard IF practice). For an intracellular granule epitope, assess permeabilisation after fixation; TPSAB1 has no transmembrane segment, but the catalog epitope is unspecified (UniProt Q15661: topology; datasheet PB10016). Match anatomical regions when comparing methods, while recognising that the selected catalog evidence documents paraffin-section chromogenic IHC rather than IF performance (datasheet PB10016).
What should I check when DAB background obscures scattered positive cells?
Inspect a no-primary control to separate secondary-reagent or chromogen background from primary-antibody staining (standard IHC practice). The selected method used 10% goat serum, a biotinylated secondary reagent, a streptavidin–biotin complex and DAB (datasheet PB10016). For that detection scheme, assess endogenous peroxidase and endogenous biotin controls as general IHC troubleshooting steps (standard IHC practice). Shorten DAB development or adjust antibody concentration only after confirming that controls retain interpretable signal (standard IHC practice). Judge faint diffuse colour against the expected selective cytoplasmic pattern in leukocyte subsets, rather than treating every brown deposit as TPSAB1-positive (HPA tissue IHC; standard IHC interpretation).
How should I quantify TPSAB1 staining across paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the tissue compartment and scoring threshold before reviewing groups, then quantify positive cells per mm² of viable tissue or the percentage of morphologically eligible cells staining positive (standard IHC practice). An H-score can summarise intensity and positive-cell fraction when staining quality supports reproducible intensity categories (standard IHC practice). Normalise counts to measured viable area, or percentages to the eligible-cell denominator, and report which denominator was used (standard IHC practice). Exclude folds, necrosis and section edges using the same rules for every specimen (standard IHC practice). Because TPSAB1 staining is selective among leukocytes, whole-section intensity may obscure differences in those cells (HPA tissue IHC).
How can I distinguish genuine TPSAB1 staining from an artefact?
Prioritise cytoplasmic signal in cells with morphology consistent with mast cells; that pattern matches the reported selective staining of probable mast cells (HPA tissue IHC). Consider extracellular deposits in context because tryptase can be released from secretory granules during activation (UniProt Q15661: subcellular location). Treat isolated membrane-only staining, necrotic areas and strong section-edge colour as findings requiring control review (UniProt Q15661: topology; standard IHC practice). Check no-primary and endogenous-peroxidase controls before attributing DAB colour to TPSAB1 (standard IHC practice). Interpret cell identity conservatively because the tissue-IHC reliability note warns that the antibody targets protein from more than one gene (HPA tissue IHC: reliability description).
Boster reagents
Best TPSAB1 / Tryptase alpha/beta-1 IHC Antibodies
Three anti-TPSAB1 antibodies have human paraffin-section IHC images (catalog image captions). Human reactivity is listed for all three; mouse and rat reactivity is also listed for A08039-1 and M08039 (catalog reactivity).
Real IHC data
Anti-Mast Cell Tryptase/TPSAB1 Antibody ®
Cat # PB10016
Real IHC data
Anti-Tryptase Alpha / Beta 1 TPSAB1 Antibody
Cat # A08039-1
Real IHC data
Anti-Mast Cell Tryptase Rabbit Monoclonal Antibody
Cat # M08039
PB10016 has IHC images from paraffin sections of human lung, breast, and intestinal cancer tissue (PB10016 image captions). A08039-1 has a paraffin-section IHC image from human tonsil, and M08039 has one from human kidney (respective image captions).
Which to pick: For human paraffin-section IHC, PB10016 offers a documented starting workflow with EDTA pH 8.0 retrieval and 1 μg/ml primary antibody; the fixative is unreported (PB10016 image caption). For mouse or rat tissue IHC, A08039-1 and monoclonal M08039 list those species, although their paraffin-section images show human tonsil and kidney, respectively; fixation is unreported in both captions (catalog reactivity and clone; respective image captions). No SKU lists IF/ICC or provides an IF image, so these data do not support an IF/ICC pick (catalog applications and IF image fields).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q15661 (TRYB1_HUMAN, Tryptase alpha/beta-1).
- Human Protein Atlas. TPSAB1 tissue IHC expression (reliability: Supported).
- Human Protein Atlas. TPSAB1 subcellular location (ICC-IF): Localized to vesicles..
- Human Protein Atlas. TPSAB1 antibody validation summary (4 antibodies).
- Relationship between numerous mast cells and early follicular development in neonatal MRL/MpJ mouse ovaries. PloS one 2013 — PMC3790711.
- Identification and Experimental Validation of Key Biomarkers for Rheumatoid Arthritis Based on Bioinformatics Analysis and Machine Learning. Journal of inflammation research 2026 — PMC13460010.
- Increased mast cell activation in eosinophilic chronic obstructive pulmonary disease. Clinical & translational immunology 2022 — PMC9512688.
- Single-cell RNA sequencing uncovers an immunosuppressive program in PDAC mast cells: tumour-driven activation, proliferation, and association with Treg recruitment. Annals of medicine 2026 — PMC13537062.
- PubMed PMID:2677049 — UniProt-cited evidence.
- PubMed PMID:2187193 — UniProt-cited evidence.
- PubMed PMID:9920877 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for TPSAB1 (Tryptase alpha/beta-1). Expected localisation Cytoplasmic staining in leukocyte subsets (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet PB10016). Validated IHC antibody: PB10016 at 0.5-1μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Bone marrow (Hematopoietic cells); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human lung cancer tissue; fixative not specified (datasheet PB10016); 2. Fixation: Image fixative and duration unreported (datasheet PB10016); 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 PB10016); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet PB10016); 8. Primary antibody: Rabbit anti-TPSAB1, 0.5-1μg/ml (datasheet PB10016); 9. Primary incubation: Overnight at 4 °C (datasheet PB10016). Watch for: Fixation: Keep fixation consistent across sections. (standard IHC practice; not target-specific); Caveat: Antibody may detect protein from more than one gene (HPA tissue IHC); Isoform / epitope: Two isoforms; mature chain starts at residue 31 (UniProt); Regulation: Degranulation releases tryptase (UniProt). Evidence: UniProt Q15661, HPA tissue IHC · Supported, HPA ICC-IF · vesicles., Datasheet PB10016, PMC3790711, PMC13460010. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/TPSAB1/immunohistochemistry-ihc-guide.