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ADAM10 / Disintegrin and metalloproteinase domain-containing protein 10 · IHC design guide
Design Immunohistochemistry for ADAM10
Plan chromogenic ADAM10 IHC on paraffin sections using an IHC-validated antibody (datasheet M00566). Score the mainly cytoplasmic tissue staining (HPA tissue IHC), while considering membrane and junctional localization (UniProt).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
ADAM10 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 | Mainly cytoplasmic (HPA tissue IHC); membrane and junctional (UniProt) | |
| Staining pattern | Cytoplasmic staining across several tissue cell types (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet M00566) | |
| Positive control | Lung+4 more · see all | |
| Negative control | Adrenal gland+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 | Mature ADAM10 can cluster at adherens junctions (UniProt) | |
| Regulation | TSPAN14 promotes cell-surface expression (UniProt) | |
| Isoform / epitope | Two isoforms; check whether the epitope is extracellular or cytoplasmic (UniProt) |
Section 1
Recommended ADAM10 IHC & IF Protocols
The catalog antibody has an IHC-P protocol (datasheet M00566); the published examples below cover hepatocellular carcinoma and pancreatic cancer tissue (PMC3829731; PMC5342537).
| Sample | Paraffin-embedded human rectum adenocarcinoma tissue; fixative not specified (datasheet M00566) |
| Fixation | Image fixative and duration unreported (datasheet M00566); 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 M00566); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet M00566) |
| Primary antibody | Rabbit monoclonal (clone AAIC-1) anti-ADAM10, 1:50 recommended; image 1:200 (datasheet M00566) |
| Primary incubation | Overnight at 4 °C (datasheet M00566) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet M00566) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | ADAM10-positive staining in macrophages of lung (HPA tissue IHC: Medium). HPA tissue profile: Mainly cytoplasmic expression in several tissues. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA at pH 8.0 for the catalog antibody (datasheet M00566); citrate at pH 6.0 is a published alternative (PMC5342537).
Section 2
What Is the Expected ADAM10 Staining Pattern?
ADAM10 spans the cell membrane and also localizes to Golgi-associated vesicles and cytoplasm (UniProt O14672 topology and localization). In tissue IHC, HPA describes mainly cytoplasmic staining and reports medium staining in lung macrophages, prostate and stomach glandular cells, and bladder urothelial cells (HPA tissue IHC). HPA rates its tissue staining Approved but notes low consistency with RNA expression (HPA tissue IHC).
| Cytoplasmic staining in lung macrophages or prostate, stomach, or seminal-vesicle glandular cells. | This fits HPA's mainly cytoplasmic tissue profile and its medium staining calls for these cells (HPA tissue IHC). Assess the named cell population rather than treating every cell in the section as an expected positive. |
| Membrane-enriched staining, possibly alongside cytoplasmic puncta, in an appropriate cell population. | ADAM10 is a membrane protein also found in Golgi-associated and clathrin-coated vesicles (UniProt O14672 localization). HPA tissue IHC describes mainly cytoplasmic staining, so membrane enrichment alone is not the sole required tissue pattern (HPA tissue IHC). |
| Predominantly nuclear staining with little cytoplasmic or membrane signal. | Nuclear localization is absent from the supplied UniProt and HPA localization records (UniProt O14672; HPA subcellular). Treat this as a possible artefact; review controls and antibody specificity before scoring it as ADAM10. |
| Strong staining across cell types that HPA calls not detected, such as adrenal glandular cells. | The distribution conflicts with that HPA observation and warrants checks for cross-reactivity or endogenous detection activity (HPA tissue IHC; standard IHC practice). A single HPA negative call does not establish universal biological absence. |
| Diffuse color throughout tissue or no signal in an HPA medium-staining cell population. | Neither result cleanly reproduces the cell-resolved HPA pattern (HPA tissue IHC). Use controls to separate nonspecific background or a detection problem from genuine sample variation (standard IHC practice). |
💡Expected ADAM10 appearanceA convincing positive is cell-resolved, mainly cytoplasmic medium staining in an HPA-listed positive population, with possible membrane accentuation; uniform tissue-wide or predominantly nuclear color is suspect (HPA tissue IHC; UniProt O14672 localization).
| Cellular compartment | The single transmembrane segment places ADAM10 at membranes, while UniProt also lists Golgi-associated vesicles (UniProt O14672 topology and localization). HPA's mainly cytoplasmic tissue pattern should guide the IHC readout (HPA tissue IHC). |
| Epitope position and processing | ADAM10 has an extracellular region at residues 20–672, a cytoplasmic tail at 694–748, and a cleaved propeptide at 20–213 (UniProt O14672). Interpret any compartment difference in light of the antibody's documented epitope, if known. |
| Shedding activity | ADAM10 sheds ectodomains of multiple membrane proteins (UniProt O14672 function). This activity does not by itself predict where a particular antibody will stain in a paraffin section; the antibody epitope and tissue result remain decisive. |
| Isoforms | UniProt lists 2 ADAM10 isoforms (UniProt O14672). The supplied evidence gives no isoform-specific tissue pattern, so avoid assigning a staining difference to an isoform without epitope or other supporting data. |
| Strength of tissue evidence | HPA rates the tissue IHC pattern Approved but reports low consistency between antibody staining and RNA expression (HPA tissue IHC). Its not detected calls are useful comparisons, not definitive absence controls. |
| Antibody validation | CAB001709 is IHC Approved; HPA050670 has no IHC status in the supplied list (HPA antibodies). Do not transfer an ICC/IF validation label to paraffin-section IHC. |
| IF/ICC Q: What localization is expected? | A: Plasma membrane and vesicles are the supported main locations; images are listed for A-431, U-251MG, and U2OS (HPA subcellular). This ICC/IF observation is separate from HPA's tissue IHC pattern. |
| Situation | Likely cause | Next action |
|---|---|---|
| No stain in lung macrophages or prostate glandular cells. | These cells have medium HPA staining, but the result may vary by sample or assay (HPA tissue IHC). | Check a section with identifiable expected cells, then review retrieval, antibody dilution, and detection controls as general IHC variables (standard IHC practice). |
| Color covers the section without clear cellular boundaries. | Diffuse color does not match HPA's cell-specific observations (HPA tissue IHC); background or endogenous detection activity may contribute (standard IHC practice). | Compare a no-primary control and inspect blocking, washes, and chromogen development (standard IHC practice). |
| Signal is mainly nuclear. | The supplied localization records emphasize membranes, vesicles, and cytoplasm (UniProt O14672; HPA subcellular). | Do not score nuclear-only color as a confirmed positive; compare controls and reassess antibody specificity (standard IHC practice). |
| Adrenal glandular cells stain strongly. | HPA calls that population not detected in tissue IHC (HPA tissue IHC). | Inspect morphology and no-primary controls for misplaced cell identification, cross-reactivity, or endogenous detection activity (standard IHC practice). |
| Membrane staining appears without a strong cytoplasmic pattern. | Membrane localization is supported, although HPA describes tissue staining as mainly cytoplasmic (UniProt O14672; HPA tissue IHC). | Record the compartment and cell type, then compare them with the antibody epitope and an HPA-listed positive population before interpreting the difference. |
| A bone-marrow section shows no hematopoietic-cell staining. | HPA calls these cells not detected, while UniProt lists bone-marrow expression without resolving this cell-level IHC result (HPA tissue IHC; UniProt O14672 tissue specificity). | Do not use this section alone to judge assay failure; check an HPA medium-staining cell population and its controls (HPA tissue IHC; standard IHC practice). |
Experiment evidence
IHC & IF images showing ADAM10 staining
M00566 · IHCIHC analysis of ADAM10 using anti-ADAM10 antibody (M00566) . ADAM10 was detected in a paraffin-embedded section of human rectum adenocarcino
M00566 · IHCIHC analysis of ADAM10 using anti-ADAM10 antibody (M00566) . ADAM10 was detected in a paraffin-embedded section of human glioblastoma tissue
A00566-2 · IHCImmunohistochemistry Validation of ADAM10 in Human Testis Immunohistochemical analysis of paraffin-embedded human testis tissue using anti-A
A00566-2 · IHCImmunohistochemistry Validation of ADAM10 in Mouse Thymus Immunohistochemical analysis of paraffin-embedded mouse thymus tissue using anti-A
A00566-2 · IFImmunofluorescence Validation of ADAM10 in MOLT4 Cells Immunofluorescent analysis of 4% paraformaldehyde-fixed MOLT4 cells labeling ADAM10 w
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000137845-ADAM10/tissue/cerebral+cortex#imid_564528
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000137845-ADAM10/tissue/colon#imid_564470
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000137845-ADAM10/tissue/liver#imid_564471
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000137845-ADAM10/tissue/kidney#imid_564456
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody CAB001709; green = ADAM10, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody CAB001709; green = ADAM10, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for ADAM10 IHC & IF
🧪Run lung first and score macrophages, which should stain at the HPA Medium level (HPA: Medium in lung macrophages). Use adrenal gland glandular cells as the negative tissue (HPA: Not detected in adrenal gland glandular cells); on the lung slide, treat staining in other cell types as background only if those cells have been independently confirmed negative for ADAM10 (HPA: lung row specifies macrophages only).
Positive control tissue: Lung (Macrophages, HPA Medium)
Negative control tissue: Adrenal gland (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show ADAM10 in A-431, U-251MG, U2OS, with annotated localisation: Vesicles (supported), Plasma membrane (supported) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only slide, a concentration-matched rabbit IgG isotype control, and ADAM10 knockout tissue or an epitope-matched peptide block as a biological specificity control (caption: rabbit anti-ADAM10; standard IHC practice). Quench endogenous peroxidase and check macrophage pigment against the no-primary slide before interpreting DAB signal in lung (HPA: lung macrophages; standard IHC practice).
⚠️Feasibility: No target-specific fixation window or fixation effect is reported in the supplied evidence, and the M00566 paraffin-section caption does not state a fixative (M00566 caption: fixative unreported). The reported IHC procedure uses heat retrieval in EDTA pH 8.0, but whether ADAM10 staining depends on that retrieval condition is unreported (M00566 caption: EDTA pH 8.0). The evidence does not establish frozen sections or IF as easier than paraffin IHC; macrophage pigment and endogenous peroxidase can complicate lung DAB interpretation (HPA: lung macrophages; standard IHC practice).
HPA tissue IHC evidence for ADAM10
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 |
|---|---|---|---|---|
| Lung | Macrophages | Medium | Protein (IHC) | HPA → |
| Prostate | Glandular cells | Medium | Protein (IHC) | HPA → |
| Seminal vesicle | Glandular cells | Medium | Protein (IHC) | HPA → |
| Stomach | Glandular cells | Medium | Protein (IHC) | HPA → |
| Urinary bladder | Urothelial cells | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adrenal gland | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Caudate | Glial cells | Not detected | Protein (IHC) | HPA → |
| Cerebellum | Cells in granular layer | Not detected | Protein (IHC) | HPA → |
| Cerebral cortex | Glial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced ADAM10 IHC Tips
Troubleshoot ADAM10 staining in paraffin sections by checking retrieval, detection controls, and whether the signal matches its documented cellular distribution.
How should I retrieve ADAM10 in paraffin sections when staining is weak?
Start with heat mediated retrieval in EDTA at pH 8.0 for paraffin sections (datasheet M00566). The documented tissue image used this retrieval before an antibody dilution of 1:200 and overnight incubation at 4°C (datasheet M00566). If staining is weak, compare a fresh section retrieved under the same conditions with a matched section given a modestly longer heating interval, then check tissue integrity and background before changing antibody concentration (standard IHC practice). Keep section thickness, detection chemistry, and imaging conditions constant so a change in signal can be attributed to retrieval (standard IHC practice).
Could fixation explain inconsistent ADAM10 staining between paraffin specimens?
The selected paraffin tissue caption does not state a fixative, so target specific fixation sensitivity for ADAM10 is unknown (datasheet M00566). Record each specimen’s fixative, fixation duration, processing history, and section age before comparing staining, because these variables can alter antigen accessibility in paraffin IHC (standard IHC practice). Run specimens with differing fixation histories in the same staining batch using EDTA retrieval at pH 8.0 and the same antibody dilution (datasheet M00566; standard IHC practice). If results remain inconsistent, report fixation history alongside the scores and avoid assigning the difference to ADAM10 abundance without an independent check (standard IHC practice).
Is cytoplasmic ADAM10 staining credible when I expect a membrane protein?
Yes: tissue IHC shows mainly cytoplasmic staining in several tissues, while ADAM10 is also documented at the cell membrane, Golgi, and cytoplasmic vesicles (HPA tissue IHC; UniProt O14672 subcellular). In paraffin sections, score membrane associated and cytoplasmic staining separately, and note whether either pattern follows intact cellular outlines rather than folds or damaged edges (standard IHC practice). The mature form is predominantly at the cell membrane and zonula adherens, whereas the immature form is mainly near cytoplasmic fibrillar structures (UniProt O14672 subcellular). A cytoplasmic DAB signal alone cannot establish which form is present or whether the enzyme is active (UniProt O14672 processing; standard IHC interpretation).
Can this stain distinguish ADAM10 isoforms or its mature form?
Do not assign an isoform from DAB staining unless the antibody’s epitope and isoform coverage have been established independently; the supplied product caption gives neither (datasheet M00566). ADAM10 has 2 listed isoforms, a propeptide at residues 20–213, and a processed chain beginning at residue 214 (UniProt O14672 isoforms and processing). Its extracellular region spans residues 20–672, whereas the cytoplasmic tail spans 694–748, so epitope position affects what a stained compartment could represent (UniProt O14672 topology). Map the documented immunogen to these regions before interpreting a difference in staining as maturation, shedding, or altered isoform expression (UniProt O14672 topology and processing; standard IHC interpretation).
How can IF help resolve an ambiguous ADAM10 IHC pattern?
Use IF as a separate, validated assay to examine whether ADAM10 signal aligns with cell outlines or intracellular puncta; supported subcellular locations include plasma membrane and vesicles (HPA subcellular). Multiplex with a marker identifying the cell population of interest, and judge overlap within that population rather than across adjacent cells (standard IF practice). Choose fluorophores in channels with low tissue autofluorescence, include unstained and single stain controls, and inspect bleed through before calling colocalisation (standard IF practice). Because the supplied antibody epitope is unspecified, choose permeabilisation only after establishing whether the recognised region is extracellular or cytoplasmic; ADAM10 spans residues 673–693 (datasheet M00566; UniProt O14672 topology; standard IF practice).
What should I adjust when ADAM10 DAB staining is diffuse?
First compare a no primary control with the stained section to identify signal from secondary reagent, endogenous peroxidase, or DAB deposition (standard IHC practice). The documented paraffin workflow used 10% goat serum blocking, an antibody dilution of 1:200, and peroxidase detection with DAB (datasheet M00566). Confirm that the peroxidase blocking step is effective, then adjust washing, antibody concentration, and DAB development one variable at a time while preserving the documented EDTA pH 8.0 retrieval (standard IHC practice; datasheet M00566). Treat broad staining that tracks tissue edges, folds, or necrosis as suspect even when genuine ADAM10 may appear cytoplasmic (standard IHC practice; HPA tissue IHC).
How should I quantify ADAM10 across paraffin tissue sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the tissue compartment and eligible cell population before scoring, then record membrane and cytoplasmic DAB signal separately (standard IHC practice; UniProt O14672 subcellular). For cell based scoring, report the percentage of positive cells and an intensity based H-score from 0–300; for focal infiltrates, positive cell density per mm² can be more informative (standard IHC practice). Normalise counts to viable tissue area or the number of eligible cells, using the same thresholds and section handling across specimens (standard IHC practice). Report the sampled regions and exclude folds, necrosis, and edge artefact so changes in tissue composition do not masquerade as ADAM10 changes (standard IHC practice).
How do I distinguish genuine ADAM10 staining from artefact?
Look for reproducible signal in intact cells with plausible membrane associated or cytoplasmic distribution; ADAM10 is documented at the membrane and in intracellular compartments (UniProt O14672 subcellular; HPA tissue IHC). Compare the cell type with the tissue reference: lung macrophages and several glandular or urothelial populations show medium staining, while the HPA profile warns of low consistency with RNA data (HPA tissue IHC). Reject patterns confined to folds, necrosis, section edges, or a positive no primary control, and investigate endogenous peroxidase before accepting diffuse DAB signal (standard IHC practice). Finally, avoid inferring protease activity or a specific processed form from staining intensity alone (UniProt O14672 function and processing; standard IHC interpretation).
Boster reagents
Best ADAM10 / Disintegrin and metalloproteinase domain-containing protein 10 IHC Antibodies
Validated anti-ADAM10 antibodies have real IHC images from human and mouse paraffin sections and IF images from MOLT4 cells and mouse/rat tissues (M00566 and A00566-2 image captions).
Real IHC data
Anti-ADAM10 Rabbit Monoclonal Antibody
Cat # M00566
Real IHC data
Anti-ADAM10 Antibody
Cat # A00566-2
M00566 has IHC images from human paraffin tumor sections and lists human, mouse and rat reactivity (M00566 IHC captions; catalog). A00566-2 has IHC images from human testis and mouse thymus, plus IF images from MOLT4 cells and mouse/rat tissues; ICC is listed as an application (A00566-2 IHC/IF captions; catalog).
Which to pick: For paraffin tissue IHC, choose M00566 when a rabbit monoclonal with human tumor-section examples fits; its caption describes EDTA retrieval but does not report the fixative (M00566 catalog; IHC captions). Choose A00566-2 for IF/ICC because both applications are listed and IF images are supplied; its IHC captions document paraffin-section human testis and mouse thymus with citrate retrieval (A00566-2 catalog; IHC/IF captions). For cross-species IHC, A00566-2 has human and mouse image examples, while its rat examples are IF; M00566 lists human, mouse and rat reactivity but supplies human IHC examples (A00566-2 IHC/IF captions; M00566 catalog and IHC captions). The selected M00566 tissue-IHC caption documents paraffin sections, but does not specify the fixative (selected-SKU IHC image M00566).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry O14672 (ADA10_HUMAN, Disintegrin and metalloproteinase domain-containing protein 10).
- Human Protein Atlas. ADAM10 tissue IHC expression (reliability: Approved).
- Human Protein Atlas. ADAM10 subcellular location (ICC-IF): Mainly localized to the plasma membrane and vesicles..
- Human Protein Atlas. ADAM10 antibody validation summary (2 antibodies).
- ADAM10 expression in gastric adenocarcinoma: Results of a curative gastrectomy cohort. Pakistan journal of medical sciences 2021 — PMC7931277.
- Upregulated expression levels of ADAM10 and EGFR and downregulated expression levels of E-cadherin in hepatocellular carcinomas. Experimental and therapeutic medicine 2013 — PMC3829731.
- Gemcitabine inhibits immune escape of pancreatic cancer by down regulating the soluble ULBP2 protein. Oncotarget 2016 — PMC5342537.
- Involvement of IRS-1 interaction with ADAM10 in the regulation of neurite extension. Journal of cellular physiology 2014 — PMC4204481.
- PubMed PMID:9305925 — UniProt-cited evidence.
- PubMed PMID:8694785 — UniProt-cited evidence.
- PubMed PMID:16572171 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for ADAM10 (Disintegrin and metalloproteinase domain-containing protein 10). Expected localisation Mainly cytoplasmic (HPA tissue IHC); membrane and junctional (UniProt). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet M00566). Validated IHC antibody: M00566 at 1:50 recommended; image 1:200 (the product's own IHC validation image is shown). Controls (HPA): positive Lung (Macrophages); negative Adrenal gland. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human rectum adenocarcinoma tissue; fixative not specified (datasheet M00566); 2. Fixation: Image fixative and duration unreported (datasheet M00566); 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 M00566); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet M00566); 8. Primary antibody: Rabbit monoclonal (clone AAIC-1) anti-ADAM10, 1:50 recommended; image 1:200 (datasheet M00566); 9. Primary incubation: Overnight at 4 °C (datasheet M00566). Watch for: Fixation: Keep fixation consistent across sections (standard IHC practice; not target-specific); Caveat: Mature ADAM10 can cluster at adherens junctions (UniProt); Isoform / epitope: Two isoforms; check whether the epitope is extracellular or cytoplasmic (UniProt); Regulation: TSPAN14 promotes cell-surface expression (UniProt). Evidence: UniProt O14672, HPA tissue IHC · Approved, HPA ICC-IF · Mainly localized to the plasma membrane and vesicles., Datasheet M00566, PMC3829731, PMC5342537. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/ADAM10/immunohistochemistry-ihc-guide.