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SH3PXD2A / SH3 and PX domain-containing protein 2A · IHC design guide
Design Immunohistochemistry for SH3PXD2A
Plan chromogenic IHC for SH3PXD2A in paraffin sections using the documented antibody dilution of 1:200 (datasheet A05241). Compare cytoplasmic and membranous staining with HPA tissue observations, bearing in mind their low concordance with RNA expression (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
SH3PXD2A 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 and membranous tissue staining (HPA tissue IHC) | |
| Staining pattern | Cytoplasmic and membranous staining in most tissues (HPA tissue IHC) | |
| Antigen retrieval | Tris-EDTA pH 9.0 HIER, heat-mediated (datasheet A05241) | |
| Positive control | Bronchus+4 more · see all | |
| Negative control | Adipose tissue+3 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 | Low concordance between staining and RNA expression (HPA tissue IHC) | |
| Regulation | RNA has low tissue specificity (HPA tissue RNA) | |
| Isoform / epitope | 3 isoforms; check epitope coverage (UniProt) |
Section 1
Recommended SH3PXD2A IHC & IF Protocols
The catalog antibody protocol uses Tris-EDTA pH 9.0 heat retrieval (datasheet A05241). The published placental IHC protocol below uses microwave retrieval in citric acid buffer (PMC7385659).
| Sample | Paraffin-embedded human uterus tissue; fixative not specified (datasheet A05241) |
| Fixation | Image fixative and duration unreported (datasheet A05241); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat retrieval: Tris-EDTA pH 9.0 (datasheet A05241); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% normal serum of the secondary host, 30 min, room temperature (standard) |
| Primary antibody | Rabbit anti-SH3PXD2A, 1:100 recommended; image 1:200 (datasheet A05241) |
| Primary incubation | Overnight at 4 °C (standard) |
| Detection | HRP-polymer secondary, DAB chromogen 5–10 min (standard) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | SH3PXD2A-positive staining in ciliated cells (ciliary rootlets) of bronchus (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic and membranous expression in most tissues. No signal in the no-primary control. |
💡Decision noteStart with Tris-EDTA pH 9.0 heat retrieval for the catalog antibody (datasheet A05241); citric acid microwave retrieval is the published alternative (PMC7385659).
Section 2
What Is the Expected SH3PXD2A Staining Pattern?
SH3PXD2A is cytoplasmic in normal cells and has no transmembrane segment (UniProt Q5TCZ1). In tissue IHC, expect cytoplasmic and membranous staining, with particularly strong staining at ciliary rootlets in ciliated cells of the bronchus, fallopian tube and nasopharynx (HPA: tissue IHC). Treat this as a qualified expectation: HPA rates its tissue staining Approved but reports low consistency with RNA expression (HPA: reliability).
| Strong, discrete staining at ciliary rootlets in ciliated epithelium. | This matches HPA's High staining in bronchus, fallopian tube and nasopharynx ciliated cells (HPA: tissue IHC). Evaluate the rootlet pattern against nearby cell morphology; brown color alone does not establish cell identity (general IHC practice). |
| Cytoplasmic and some membranous staining across several tissue compartments. | This fits HPA's broad tissue profile and UniProt's cytoplasmic localisation (HPA: tissue IHC; UniProt Q5TCZ1). Membranous staining by itself does not establish that SH3PXD2A spans the membrane: UniProt annotates no transmembrane segment (UniProt Q5TCZ1 topology). |
| Predominantly nuclear signal, with little cytoplasmic or ciliary-rootlet staining. | A nuclear-only pattern conflicts with the reported tissue and subcellular patterns (HPA: tissue IHC and ICC-IF; UniProt Q5TCZ1). Treat it as suspect staining and check the antibody and detection controls before assigning it to SH3PXD2A (general IHC practice). |
| Strong staining in adipocytes, duodenal glandular cells or lymph-node germinal-center cells. | These cell populations were Not detected in HPA tissue IHC (HPA: tissue IHC). Unexpected signal may reflect cross-reactivity or endogenous detection activity; compare matched controls and tissue morphology before calling it specific (general IHC practice). |
| Diffuse stain across the section, or no signal in ciliated cells expected to be positive. | Diffuse stain obscures the reported rootlet pattern; absent signal in an HPA High cell population leaves the run uninformative (HPA: tissue IHC). Examine background and positive controls before interpreting a negative specimen (general IHC practice). |
💡Expected SH3PXD2A appearanceCall a result positive when ciliated cells show High, discrete ciliary-rootlet staining with a compatible cytoplasmic or membranous pattern; predominantly nuclear signal or uniform background is suspect (HPA: tissue IHC; UniProt Q5TCZ1; general IHC practice).
| Cell type and tissue | HPA reports High ciliary-rootlet staining in bronchus, fallopian tube and nasopharynx; Medium staining in adrenal and appendix glandular cells; and Not detected in adipocytes (HPA: tissue IHC). Score the named cell population rather than treating an entire section as uniformly positive or negative. |
| Compartment and topology | UniProt places SH3PXD2A in cytoplasm and, in SRC-transformed cells, podosomes; it annotates no transmembrane segment (UniProt Q5TCZ1). HPA describes cytoplasmic and membranous tissue staining (HPA: tissue IHC). A routine tissue section cannot establish podosome localisation from a brown punctum alone. |
| Antibody evidence | HPA rates tissue IHC Approved, while reporting low consistency between antibody staining and RNA expression (HPA: reliability). HPA037922 and HPA037923 each have Approved IHC status; the supplied antibody list gives no IHC Enhanced status (HPA: antibodies). Interpret unexpected cells conservatively. |
| IF/ICC question: where should signal appear? | HPA reports cytosol and primary cilium transition zone as approved ICC-IF locations (HPA: subcellular). That observation can inform compartment interpretation, but it is an ICC-IF result; use the tissue IHC record for intensity expectations in paraffin sections (HPA: tissue IHC). |
| Isoforms and epitope | UniProt lists 3 isoforms and a full-length chain of residues 1–1133 (UniProt Q5TCZ1). The supplied evidence gives no antibody epitope or isoform coverage, so a staining difference cannot be assigned to a particular isoform or to epitope loss. |
| Situation | Likely cause | Next action |
|---|---|---|
| No stain at ciliary rootlets in a bronchus positive-control section. | The expected High cell population is absent from the readout (HPA: bronchus); the supplied sources do not identify a target-specific fixation or retrieval cause. | Confirm ciliated cells are present, then review the run's positive control, antibody application, retrieval and detection steps as general IHC checks; avoid calling the specimen negative until the control works. |
| Whole section is diffusely brown, including areas outside recognizable cells. | Non-specific background or detection chemistry may obscure the cell-specific pattern (general IHC practice); HPA's rootlet observation cannot be assessed through uniform stain (HPA: tissue IHC). | Compare the negative reagent control and examine blocking, washes and chromogen development using standard IHC controls; then reassess ciliary rootlets and cell outlines. |
| Stain is concentrated in nuclei. | The pattern conflicts with cytoplasmic tissue staining and the approved cytosol ICC-IF location (HPA: tissue IHC and subcellular; UniProt Q5TCZ1). | Check the negative control, tissue morphology and antibody specificity before scoring it as SH3PXD2A; retain a cytoplasmic positive control in the same run (general IHC practice). |
| Adipocytes or germinal-center cells stain strongly. | Both populations are Not detected in HPA tissue IHC (HPA: adipose tissue and lymph node); cross-reactivity or endogenous detection activity is possible (general IHC practice). | Confirm the cell type on the counterstain, compare detection controls and repeat with an independently validated antibody if available; report the conflict with HPA rather than silently recoding it as expected. |
| Low staining in lung macrophages is read as a failed stain. | HPA records lung macrophages as Low, while ciliary rootlets in bronchus are High (HPA: tissue IHC); expected intensity depends on the cell population. | Use a High-staining ciliated-cell control to judge run performance, then score macrophages within their own tissue context rather than requiring rootlet-level intensity. |
| A tissue pattern differs from RNA expression or from another antibody. | HPA explicitly reports low consistency with RNA despite an Approved IHC reliability rating (HPA: reliability); the supplied antibody records show Approved IHC for HPA037922 and HPA037923 (HPA: antibodies). | Document the antibody and the specific cell populations scored, compare controls and independent staining where available, and keep any unresolved discrepancy visible in the interpretation. |
Experiment evidence
IHC & IF images showing SH3PXD2A staining
A05241 · IHCImmunohistochemical analysis of paraffin-embedded human uterus. 1, Antibody was diluted at 1:200 (4° overnight). 2, Tris-EDTA, pH9.0 was use
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000107957-SH3PXD2A/tissue/cerebral+cortex#imid_9234648
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000107957-SH3PXD2A/tissue/colon#imid_9234574
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000107957-SH3PXD2A/tissue/liver#imid_9234583
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000107957-SH3PXD2A/tissue/kidney#imid_9234576
HPA · ICC-IF ASC52teloHuman Protein Atlas immunofluorescence, ASC52telo, antibody HPA037923; green = SH3PXD2A, blue = nucleus, red = microtubules.
HPA · ICC-IF ASC52teloHuman Protein Atlas immunofluorescence, ASC52telo, antibody HPA037923; green = SH3PXD2A, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for SH3PXD2A IHC & IF
🧪Run bronchus first and look for staining in ciliated-cell ciliary rootlets (HPA: High in bronchus ciliated cells). Run duodenum as a negative tissue, focusing on glandular cells (HPA: Not detected in duodenum glandular cells); on the bronchus slide, use adjacent non-ciliated cells to assess background rather than assuming they are target-negative (HPA: High signal is assigned to ciliated-cell ciliary rootlets).
Positive control tissue: Bronchus (Ciliated cells (ciliary rootlets), HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show SH3PXD2A in ASC52telo, RPTEC/TERT1, hTERT-RPE1 (serum starved), with annotated localisation: Primary cilium transition zone (approved), Cytosol (approved) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control and a concentration-matched irrelevant antibody control matched to the primary antibody’s host and isotype or clonality (standard IHC practice). Confirm specificity with target knockout material or immunizing-peptide competition if available, and quench endogenous peroxidase in bronchus sections before chromogenic detection (standard IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported in the supplied evidence, and the selected A05241 paraffin-section caption does not report a fixative (A05241 tissue-IHC caption). The caption uses Tris-EDTA at pH 9.0 for retrieval in uterus; retrieval conditions for bronchus need checking on the chosen sections (A05241 tissue-IHC caption: uterus, Tris-EDTA pH 9.0). The evidence does not establish that frozen sections or IF are easier; for bronchus IHC, assess ciliary-rootlet staining with section orientation and luminal staining artefact in mind (HPA: High in bronchus ciliated-cell ciliary rootlets; standard IHC practice).
HPA tissue IHC evidence for SH3PXD2A
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — Low consistency between antibody staining and RNA expression data.). 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 |
|---|---|---|---|---|
| Bronchus | Ciliated cells (ciliary rootlets) | High | Protein (IHC) | HPA → |
| Fallopian tube | Ciliated cells (ciliary rootlets) | High | Protein (IHC) | HPA → |
| Nasopharynx | Ciliated cells (ciliary rootlets) | High | Protein (IHC) | HPA → |
| Adrenal gland | Glandular cells | Medium | Protein (IHC) | HPA → |
| Appendix | Glandular cells | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Duodenum | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Lymph node | Germinal center cells | Not detected | Protein (IHC) | HPA → |
| Parathyroid gland | Glandular cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced SH3PXD2A IHC Tips
Troubleshoot SH3PXD2A staining in paraffin sections by checking retrieval, cellular compartment, controls and scoring before interpreting signal as podosome activity (UniProt Q5TCZ1).
Which retrieval conditions should I try first for weak SH3PXD2A staining?
Start with heat-mediated retrieval in Tris-EDTA at pH 9.0 (datasheet A05241). The paraffin-section example used that buffer with antibody at 1:200 overnight at 4°C, so keep those conditions constant while assessing retrieval (datasheet A05241). If staining remains weak, compare a shorter or longer heating interval on adjacent sections, allowing each section to cool consistently before washing (standard IHC practice). Check preservation of morphology and include a no-primary control, since stronger chromogen alone does not establish specific staining (standard IHC practice). Record the cell types and compartments that change: SH3PXD2A has cytoplasmic and cell-projection localization (UniProt Q5TCZ1).
How should I handle fixation when SH3PXD2A staining varies between specimens?
Target-specific sensitivity to fixation is unknown from the supplied evidence; the paraffin-section caption does not state a fixative (datasheet A05241). Record each specimen’s fixative and fixation duration, then compare sections processed together with the same Tris-EDTA retrieval at pH 9.0 (datasheet A05241; standard IHC practice). Keep section thickness, primary incubation and chromogen development consistent so processing differences remain interpretable (standard IHC practice). The illustrated primary incubation was 1:200 overnight at 4°C, but that example alone cannot establish fixation tolerance (datasheet A05241). Assess morphology and background alongside cellular staining before attributing variation to antigen loss (standard IHC practice).
What staining pattern is plausible for SH3PXD2A in paraffin sections?
Expect cellular staining compatible with a cytoplasmic protein; SH3PXD2A lacks a transmembrane segment and is annotated in cytoplasm and podosomes (UniProt Q5TCZ1). Tissue IHC reports cytoplasmic and membranous expression in most tissues, but its staining has low consistency with RNA data (HPA tissue IHC). High staining at ciliary rootlets is reported in ciliated cells of bronchus, fallopian tube and nasopharynx (HPA tissue IHC). Examine these structures at sufficient magnification and compare the same cell type across adjacent sections before calling a fine apical signal specific (standard IHC practice). A sharply nuclear-only pattern warrants antibody and detection controls before biological interpretation (UniProt Q5TCZ1; standard IHC practice).
Could isoforms or epitope placement explain inconsistent SH3PXD2A staining?
SH3PXD2A has 3 listed isoforms and a PX domain plus 5 SH3 domains across its 1,133 residues (UniProt Q5TCZ1). Confirm the catalog antibody’s immunogen coordinates and whether its epitope is shared by the isoforms before comparing specimens (standard IHC practice). The supplied caption documents staining conditions but provides no epitope coordinates or isoform coverage (datasheet A05241). If staining differs between specimens, first repeat matched retrieval at pH 9.0 and review morphology and controls (datasheet A05241; standard IHC practice). Phosphorylated residues are annotated, but the supplied evidence does not show that their state changes antibody binding (UniProt Q5TCZ1).
How can I check SH3PXD2A localization by IF alongside paraffin-section IHC?
For a separate IF experiment, multiplex SH3PXD2A with a marker identifying the expected cell type, such as a ciliated-cell marker when examining reported ciliary-rootlet staining (HPA tissue IHC; standard IF practice). Select fluorophores after measuring tissue autofluorescence in an unstained control, and favor a detection channel with clear separation from that background (standard IF practice). SH3PXD2A is annotated in cytosol and the primary cilium transition zone; select permeabilisation according to epitope accessibility and verify it against a no-primary control (HPA subcellular; standard IF practice). Compare cellular location with the chromogenic result, while treating differences in processing and detection as possible explanations for discordance (standard IHC/IF practice).
How do I reduce diffuse brown background without losing SH3PXD2A signal?
Start with a no-primary control and inspect whether brown deposit tracks tissue edges, damaged areas or structures outside the expected cellular pattern (standard IHC practice; UniProt Q5TCZ1). For peroxidase-based chromogenic detection, use an appropriate endogenous peroxidase block and check its effectiveness in the control section (standard IHC practice). Titrate primary antibody around the illustrated 1:200 condition, holding retrieval at pH 9.0 and development time constant during the comparison (datasheet A05241; standard IHC practice). Optimize blocking and washes if diffuse staining persists, then judge signal against morphology and cell-specific distribution (standard IHC practice). The reported tissue staining has low RNA concordance, so broad positivity deserves particular caution (HPA tissue IHC).
What should I score when comparing SH3PXD2A staining across paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the compartment and cell population before scoring; reported tissue patterns include cytoplasmic and membranous staining and prominent ciliary rootlets in specified ciliated cells (HPA tissue IHC). For a broad cellular signal, record percentage-positive cells and intensity or calculate an H-score from 0–300 using intensity-weighted percentages (standard IHC practice). For a restricted structure such as ciliary rootlets, record positive profiles per mm² within a consistently defined epithelial region (HPA tissue IHC; standard IHC practice). Normalize counts to the relevant cells or measured tissue area, and exclude folds, necrosis and detached edges by a rule set before review (standard IHC practice). Use identical staining and image-analysis settings across compared sections (standard IHC practice).
When should I question an apparent SH3PXD2A-positive focus?
Check that the focus lies in an interpretable cell and compartment: SH3PXD2A is annotated in cytoplasm and podosomes, while tissue IHC also reports ciliary-rootlet staining in certain ciliated cells (UniProt Q5TCZ1; HPA tissue IHC). Compare neighboring intact cells and a no-primary section when signal is confined to an edge, necrotic area or endogenous-enzyme-rich region (standard IHC practice). A nuclear-only focus or uniform deposit across unrelated structures needs further validation before attribution to SH3PXD2A (UniProt Q5TCZ1; standard IHC practice). Even plausible staining does not, by itself, demonstrate invadopodia activity or matrix degradation, and the reported tissue IHC has low consistency with RNA expression (UniProt Q5TCZ1; HPA tissue IHC).
Boster reagents
Best SH3PXD2A / SH3 and PX domain-containing protein 2A IHC Antibodies
The catalog includes an IHC image of paraffin-embedded human uterus (A05241 image caption), IF/ICC applications (catalog: application lists), and listed human, mouse, and rat reactivity (catalog: reactivity lists).
Real IHC data
Anti-Fish SH3PXD2A Antibody
Cat # A05241
A05241 has an IHC image of paraffin-embedded human uterus and lists IHC and IF applications with human and mouse reactivity (A05241 image caption; catalog: applications and reactivity). A05241-1 lists IHC, IF, and ICC applications with human, mouse, and rat reactivity; no IHC or IF image is supplied (catalog: A05241-1 applications, reactivity, and image fields).
Which to pick: Choose A05241 for tissue IHC when a documented paraffin-section example is useful: its image shows human uterus after Tris-EDTA retrieval at pH 9.0 and primary antibody at 1:200 overnight at 4°C (A05241 IHC image caption). Choose A05241-1 for IF/ICC or rat samples because it lists both applications and rat reactivity and is described as polyclonal (catalog: A05241-1 applications, reactivity, and dilution text). The fixative is unreported for the A05241 paraffin-section image, and A05241-1 has no supplied processing image (A05241 IHC image caption; catalog: A05241-1 image fields).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q5TCZ1 (SPD2A_HUMAN, SH3 and PX domain-containing protein 2A).
- Human Protein Atlas. SH3PXD2A tissue IHC expression (reliability: Approved).
- Human Protein Atlas. SH3PXD2A subcellular location (ICC-IF): Localized to the cytosol and primary cilium transition zone..
- Human Protein Atlas. SH3PXD2A antibody validation summary (2 antibodies).
- Association of lncRNA SH3PXD2A-AS1 with preeclampsia and its function in invasion and migration of placental trophoblast cells. Cell death & disease 2020 — PMC7385659.
- The role and regulation of Rab40b-Tks5 complex during invadopodia formation and cancer cell invasion. Journal of cell science 2016 — PMC5201011.
- PubMed PMID:9455477 — UniProt-cited evidence.
- PubMed PMID:15164054 — UniProt-cited evidence.
- PubMed PMID:12615925 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for SH3PXD2A (SH3 and PX domain-containing protein 2A). Expected localisation Cytoplasmic and membranous tissue staining (HPA tissue IHC). Antigen retrieval: Tris-EDTA pH 9.0 HIER, heat-mediated (datasheet A05241). Validated IHC antibody: A05241 at 1:100 recommended; image 1:200 (the product's own IHC validation image is shown). Controls (HPA): positive Bronchus (Ciliated cells (ciliary rootlets)); negative Adipose tissue. IHC workflow; specimen preparation unspecified. Protocol steps: 1. Sample: Paraffin-embedded human uterus tissue; fixative not specified (datasheet A05241); 2. Fixation: Image fixative and duration unreported (datasheet A05241); 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: Tris-EDTA pH 9.0 (datasheet A05241); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% normal serum of the secondary host, 30 min , room temperature (standard); 8. Primary antibody: Rabbit anti-SH3PXD2A, 1:100 recommended; image 1:200 (datasheet A05241); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep fixation consistent across sections (standard IHC practice; not target-specific); Caveat: Low concordance between staining and RNA expression (HPA tissue IHC); Isoform / epitope: 3 isoforms; check epitope coverage (UniProt); Regulation: RNA has low tissue specificity (HPA tissue RNA). Evidence: UniProt Q5TCZ1, HPA tissue IHC · Approved, HPA ICC-IF · cytosol and primary cilium transition zone., Datasheet A05241, PMC7385659. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/SH3PXD2A/immunohistochemistry-ihc-guide.