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MMP14 / Matrix metalloproteinase-14 · IHC design guide
Design Immunohistochemistry for MMP14
Plan chromogenic MMP14 IHC in paraffin sections around the observed cytoplasmic tissue pattern (HPA tissue IHC). Start with the documented 1:25 A00656 dilution (datasheet A00656), then score staining by cell type and compartment in light of MMP14’s membrane topology (UniProt).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
MMP14 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 tissues (HPA tissue IHC) | |
| Staining pattern | Cytoplasmic staining in several tissue cell types (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6 HIER, heat-mediated (datasheet A00656) | |
| Positive control | Adipose tissue+4 more · see all | |
| Negative control | Bone marrow+4 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Formaldehyde-fixed paraffin sections are documented (selected-SKU IHC image A00656) | |
| Caveat | Staining has medium consistency with RNA expression (HPA tissue IHC) | |
| Regulation | Specific expression regulation is not reported (UniProt) | |
| Isoform / epitope | 0 isoforms annotated; extracellular versus cytoplasmic epitope matters (UniProt) |
Section 1
Recommended MMP14 IHC & IF Protocols
The catalog antibody’s IHC-P protocol is presented alongside published protocols for human pleural mesothelioma (PMC2737627) and rat bone (PMC13101557).
| Sample | Formaldehyde-fixed, paraffin-embedded human lung adenocarcinoma tissue (datasheet A00656) |
| Fixation | Image formalin-fixed; duration unreported (datasheet A00656); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat retrieval: Citrate pH 6 (datasheet A00656); 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-MMP14, 1:25 (datasheet A00656) |
| Primary incubation | 1 hours at 37°C (datasheet A00656) |
| Detection | HRP-polymer secondary, DAB chromogen 5–10 min (standard) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | MMP14-positive staining in adipocytes of adipose tissue (HPA tissue IHC: Medium). HPA tissue profile: Cytoplasmic expression in several tissues. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated citrate retrieval at pH 6 for the catalog antibody (datasheet A00656); the mesothelioma protocol also used citrate at pH 6.0 (PMC2737627).
Section 2
What Is the Expected MMP14 Staining Pattern?
MMP14 is a membrane protein with an extracellular region and a short cytoplasmic tail; UniProt also lists cytoplasmic localization (UniProt P50281 topology and subcellular location). On paraffin sections, expect chiefly cytoplasmic staining in selected cells, including endometrial stromal cells and bronchial basal cells at medium intensity (HPA tissue IHC). Interpret this pattern with care: HPA rates its tissue IHC Approved, with medium consistency against RNA expression (HPA tissue IHC).
| Selected endometrial stromal cells or bronchial basal cells show moderate cytoplasmic chromogen, with defined cell boundaries. | This fits the reported medium staining in those cell populations and HPA's overall cytoplasmic tissue profile (HPA tissue IHC). A membrane accent is plausible from MMP14 topology, but the supplied tissue IHC profile does not require a crisp membrane rim (UniProt P50281 topology; HPA tissue IHC). |
| The dominant signal is confined to nuclei, with little corresponding cytoplasmic or cell surface staining. | Treat a nuclear-only pattern as suspect: the supplied localization records list cell membrane and cytoplasm, while HPA describes cytoplasmic tissue staining (UniProt P50281 subcellular location; HPA tissue IHC). Check a known-positive cell population and controls before assigning this signal to MMP14 (general IHC practice). |
| Hematopoietic cells throughout bone marrow, or germinal center cells in lymph node, stain as strongly as the chosen positive cells. | Those specific populations were reported as not detected in HPA tissue IHC; widespread staining there raises concern for nonspecific binding or endogenous chromogenic activity (HPA tissue IHC; general IHC practice). HPA's result is an assay observation, not proof that every cell in those tissues lacks MMP14. |
| Brown color spreads across extracellular spaces, section edges, or unrelated cell populations without clear cellular structure. | An indiscriminate pattern cannot be assigned to the reported cytoplasmic distribution (HPA tissue IHC). Inspect a primary-antibody-omission control for detection-system background, and review blocking, washes, and chromogen development (general IHC practice). |
| A technically intact section shows no signal in endometrial stromal cells or bronchial basal cells. | These are reported medium-positive populations, so an entirely blank result warrants assay troubleshooting (HPA tissue IHC). Their HPA level is an observation under that assay, not a guaranteed intensity with another antibody or workflow; compare tissue integrity and assay controls before calling MMP14 absent (general IHC practice). |
💡Expected MMP14 appearanceCall an IHC result consistent with MMP14 when selected reported-positive cells show interpretable medium cytoplasmic staining, possibly with a membrane accent; uniform staining of unrelated cells or nuclear-only color is suspect (HPA tissue IHC; UniProt P50281 topology).
| Topology and epitope side | MMP14 spans residues 542–562; residues 112–541 are extracellular and 563–582 cytoplasmic (UniProt P50281 topology). The supplied record does not identify an antibody epitope, so topology alone cannot predict which staining component that antibody will emphasize. |
| Precursor processing | The signal peptide occupies residues 1–20, the propeptide 21–111, and the annotated mature chain 112–582 (UniProt P50281 processing). If an antibody's epitope becomes known, compare it with these regions before interpreting differences between precursor and mature protein; no epitope is supplied here. |
| Strength of tissue evidence | HPA calls the tissue IHC profile Approved and describes medium consistency between antibody staining and RNA data (HPA tissue IHC). Its medium and not-detected labels describe particular cell populations; they should guide comparator selection without becoming universal expression rules (HPA tissue IHC). |
| IF/ICC Q&A: should the IHC pattern be copied into IF? | No. On the separate ICC-IF record, MMP14 is mainly cytosolic, with additional intermediate-filament localization; both locations are approved (HPA subcellular ICC-IF). Those observations help interpret IF images but do not specify a paraffin IHC staining requirement or an IF protocol. |
| Situation | Likely cause | Next action |
|---|---|---|
| Positive comparator is blank. | The tested stain may lack detectable signal, or the selected section may not contain the reported positive cells (HPA tissue IHC; general IHC practice). | Locate endometrial stromal or bronchial basal cells on the section, then check antibody and detection controls before interpreting the study tissue (HPA tissue IHC; general IHC practice). |
| All cells appear brown. | Nonspecific detection or excessive chromogen development can obscure the selective HPA pattern (HPA tissue IHC; general IHC practice). | Compare a primary-antibody-omission control; review blocking, washing, and development time using general IHC practice. |
| The signal appears exclusively nuclear. | This conflicts with the supplied membrane and cytoplasmic localization evidence (UniProt P50281 subcellular location; HPA tissue IHC). | Recheck morphology and controls, then repeat interpretation against a reported-positive cell population (HPA tissue IHC; general IHC practice). |
| Bone marrow hematopoietic cells or lymph node germinal centers stain strongly. | That conflicts with HPA's not-detected observations for those cell populations; nonspecific antibody or detection activity is possible (HPA tissue IHC; general IHC practice). | Run an omission control and compare staining with a reported medium-positive population on a separate control section (HPA tissue IHC; general IHC practice). |
| A different antibody gives a different balance of cytoplasmic and membrane signal. | The supplied antibodies have IHC Approved status, but their epitopes are unspecified; topology cannot establish equivalence (HPA antibodies; UniProt P50281 topology). | Compare each antibody's own controlled pattern with the HPA tissue profile; avoid inferring epitope recognition from staining alone (HPA tissue IHC; general IHC practice). |
| Retrieval changes the amount of visible stain. | The supplied UniProt and HPA records provide no MMP14-specific fixation or retrieval sensitivity evidence. | Treat retrieval as an assay variable: compare controlled sections and retain the condition that yields interpretable cellular staining with low background (general IHC practice). |
Experiment evidence
IHC & IF images showing MMP14 staining
A00656 · IHCA00656 staining MMP14 in human lung adenocarcinoma tissue sections by Immunohistochemistry (IHC-P -paraformaldehyde-fixed, paraffin-embedded
PA2147 · IHCAnti-MMP14 antibody, PA2147, IHC(P) IHC(P): Human Intestinal Cancer Tissue
PA2147 · IHCAnti-MMP14 antibody, PA2147, IHC(P) IHC(P): Human Placenta Tissue
M00656 · IHCImmunohistochemical analysis of paraffin-embedded human kdieny, using MMP14 Antibody .
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000157227-MMP14/tissue/cerebral+cortex#imid_2763396
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000157227-MMP14/tissue/colon#imid_2763338
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000157227-MMP14/tissue/liver#imid_2763331
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000157227-MMP14/tissue/kidney#imid_2763324
![HPA ICC-IF BJ [Human fibroblast] — MMP14 (HPA051432)](https://images.proteinatlas.org/51432/1658_G1_1_blue_red_green.jpg)
HPA · ICC-IF BJ [Human fibroblast]Human Protein Atlas immunofluorescence, BJ [Human fibroblast], antibody HPA051432; green = MMP14, blue = nucleus, red = microtubules.
![HPA ICC-IF BJ [Human fibroblast] — MMP14 (HPA051432)](https://images.proteinatlas.org/51432/1658_G1_3_blue_red_green.jpg)
HPA · ICC-IF BJ [Human fibroblast]Human Protein Atlas immunofluorescence, BJ [Human fibroblast], antibody HPA051432; green = MMP14, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for MMP14 IHC & IF
🧪Run bronchus first and look for staining in basal cells, which HPA scores Medium (HPA: bronchus, basal cells). Use bone marrow hematopoietic cells as the negative tissue (HPA: bone marrow, hematopoietic cells, Not detected); on the bronchus slide, assess cells outside the basal compartment for little or no background staining without assuming every such cell is MMP14-negative.
Positive control tissue: Adipose tissue (Adipocytes, HPA Medium)
Negative control tissue: Bone marrow (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show MMP14 in BJ [Human fibroblast], HaCaT, U2OS, with annotated localisation: Cytosol (approved) (HPA subcellular).
Technical controls: Include no-primary (secondary-only) and host-species- and clonality-matched isotype controls, plus an MMP14 knockout sample or peptide-block control if the immunizing peptide is available (standard IHC controls). Quench endogenous peroxidase in bronchus sections and block endogenous biotin if using the caption’s biotinylated secondary antibody (A00656 tissue-IHC caption: biotinylated secondary).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected A00656 tissue-IHC caption’s fixative field is unreported. The caption uses heat-mediated citrate retrieval at pH 6, but does not establish that retrieval is required for every MMP14 antibody (A00656 tissue-IHC caption: citrate retrieval). Frozen-section performance and relative ease of IF are unreported; for bronchus IHC, endogenous peroxidase can produce background and should be quenched (standard IHC practice).
HPA tissue IHC evidence for MMP14
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — Medium 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 |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Medium | Protein (IHC) | HPA → |
| Breast | Adipocytes | Medium | Protein (IHC) | HPA → |
| Bronchus | Basal cells | Medium | Protein (IHC) | HPA → |
| Cervix | Glandular cells | Medium | Protein (IHC) | HPA → |
| Duodenum | Glandular cells | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Bone marrow | Hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Cerebellum | Cells in granular layer | Not detected | Protein (IHC) | HPA → |
| Lymph node | Germinal center cells | Not detected | Protein (IHC) | HPA → |
| Salivary gland | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Spleen | Cells in red pulp | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced MMP14 IHC Tips
Troubleshoot MMP14 staining in paraffin sections by checking retrieval, compartment, cell identity, and controls before comparing chromogenic signal across samples.
What should I change when MMP14 staining is weak after antigen retrieval?
Start with heat-mediated citrate retrieval at pH 6 for paraffin sections (datasheet A00656). The selected IHC-P example uses this retrieval before a 1:25 primary incubation for 1 hour at 37°C (caption A00656). If signal is weak, compare retrieval duration on adjacent sections while keeping antibody dilution, detection, and development time fixed (standard IHC practice). Check a positive reference alongside each condition; stromal cells in colon, breast, and head and neck are reported expression sites, but their staining intensity in your material still needs validation (UniProt P50281 tissue specificity; standard IHC practice).
Could fixation explain weak or uneven MMP14 staining?
MMP14-specific fixation sensitivity is unknown from the supplied evidence, so do not assign weak staining to a particular fixative without a controlled comparison. Record fixation method, duration, and tissue thickness for each paraffin sample, then compare matched sections processed through the same retrieval and detection steps (standard IHC practice). Uneven fixation or processing can produce regional differences in antigen accessibility; review section morphology and exclude damaged edges before interpreting intensity (standard IHC practice). Include a reference section in every run and change only one preparation variable at a time, because differences in fixation and retrieval can otherwise be confounded (standard IHC practice).
Should MMP14 appear at the cell surface or in the cytoplasm?
Expect compartment-dependent staining: MMP14 has an extracellular region at residues 112–541, a transmembrane segment at 542–562, and a cytoplasmic tail at 563–582 (UniProt P50281 topology). Cytoplasmic tissue staining is reported, while cytosol is the approved main location in cell imaging (HPA tissue IHC; HPA subcellular). Score membranous and cytoplasmic patterns separately, using cell boundaries and morphology to distinguish them from diffuse chromogen spread (standard IHC practice). Interpret staining in its cell context because expression is reported in stromal cells of colon, breast, and head and neck, and in lung tumors (UniProt P50281 tissue specificity).
How does an unmapped epitope affect interpretation of MMP14 staining?
The supplied record annotates 0 isoforms, but it does not map the catalog antibody’s epitope; do not infer which molecular region this stain detects (UniProt P50281 isoforms; supplied antibody evidence). MMP14 is processed from a signal sequence at residues 1–20 and propeptide at 21–111 to a chain spanning 112–582 (UniProt P50281 processing). The extracellular region and cytoplasmic tail therefore pose different accessibility questions, which require epitope information before assigning a staining pattern to one side of the membrane (UniProt P50281 topology; standard IHC practice). A phosphotyrosine is annotated at residue 399; this alone does not establish that the antibody is modification-specific (UniProt P50281 modified residues; supplied antibody evidence).
How should I check MMP14 localisation in a multiplex IF experiment?
Pair MMP14 with a validated marker for the cell population being examined, and inspect the channels separately before calling co-localisation (standard IF practice). Choose a fluorophore and filter set that separates the MMP14 channel from measured tissue autofluorescence, using single-stain and unstained controls (standard IF practice). Because the antibody epitope is unmapped, test permeabilised and non-permeabilised conditions when asking whether signal is accessible at the cell surface; the extracellular region spans 112–541 and the cytoplasmic tail 563–582 (supplied antibody evidence; UniProt P50281 topology; standard IF practice). Cytosolic and intermediate-filament localisation has been reported in cell imaging, so assess intracellular signal without assuming that every punctum is surface MMP14 (HPA subcellular; standard IF practice).
How can I distinguish MMP14 signal from chromogenic background?
Run a no-primary control through the same secondary and chromogen steps to reveal detection-system background, and compare it with a positive reference section (standard IHC practice). The selected example used a biotinylated secondary after 3% BSA blocking for 0.5 hour, so check for background associated with the biotin-based detection system (caption A00656; standard IHC practice). Include a peroxidase block before DAB development and assess any residual endogenous enzyme signal in the no-primary control (standard IHC practice). If diffuse staining persists, optimise blocking, primary dilution, washing, and chromogen development one variable at a time while retaining the same retrieval condition (standard IHC practice).
How should I quantify MMP14 staining across paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the analysis region and cell population before scoring, then report the percentage of positive cells and an intensity-weighted H-score using identical thresholds across slides (standard IHC practice). Score membranous and cytoplasmic signal separately because MMP14 has a transmembrane segment and cytoplasmic tail, while tissue IHC reports cytoplasmic expression (UniProt P50281 topology; HPA tissue IHC). Normalise positive-cell counts to the number of eligible cells in each region, or report positive-cell density per mm² of evaluable tissue (standard IHC practice). Exclude folds, necrosis, and section edges, and keep retrieval, chromogen development, imaging, and scoring rules consistent across comparison groups (standard IHC practice).
When is an apparent MMP14-positive area likely to be an artefact?
Treat staining confined to section edges, folds, or necrotic material as suspect, particularly when similar colour appears in the no-primary control (standard IHC practice). Check whether signal follows identifiable cells and a plausible compartment: MMP14 is membrane-associated with a cytoplasmic tail, and cytoplasmic tissue staining is reported (UniProt P50281 topology; HPA tissue IHC). An unexpected cell population warrants validation against morphology and controls; HPA reports medium staining in bronchial basal cells but no detection in lymph-node germinal-center cells (HPA tissue IHC; standard IHC practice). Residual endogenous peroxidase can mimic DAB signal, so compare peroxidase-blocked and no-primary sections before calling isolated deposits positive (standard IHC practice).
Boster reagents
Best MMP14 / Matrix metalloproteinase-14 IHC Antibodies
All three antibodies list human, mouse and rat reactivity (catalog). Their IHC figures show human lung adenocarcinoma, intestinal cancer, placenta and kidney, plus rat cardiac muscle (figure captions).
Real IHC data
Anti-MMP14 Antibody (N-term)
Cat # A00656
Real IHC data
Anti-Matrix metalloproteinase-14 MMP14 Antibody ®
Cat # PA2147
Real IHC data
Anti-MMP14/Mt1 Mmp Rabbit Monoclonal Antibody
Cat # M00656
A00656 shows IHC-P in formaldehyde-fixed, paraffin-embedded human lung adenocarcinoma (A00656 caption); M00656 shows IHC in paraffin-embedded human kidney, with fixative unreported (M00656 caption). PA2147 shows IHC(P) in human intestinal cancer and placenta and rat cardiac muscle; fixative is unreported (PA2147 captions).
Which to pick: For tissue IHC, choose A00656 when its documented formaldehyde fixation, citrate retrieval and 1:25 dilution fit your sections (A00656 caption); PA2147 provides human intestinal cancer, placenta and rat cardiac muscle examples, with fixative unreported (PA2147 captions). For IF/ICC, M00656 is the only listed option, with a 1:50 IF dilution and rabbit monoclonal clone BFD-13 (M00656 catalog); its IHC figure shows paraffin-embedded human kidney, with fixative unreported (M00656 caption). For cross-species planning, all three list human, mouse and rat reactivity (catalog), while PA2147 has the rat IHC example (PA2147 caption).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P50281 (MMP14_HUMAN, Matrix metalloproteinase-14).
- Human Protein Atlas. MMP14 tissue IHC expression (reliability: Approved).
- Human Protein Atlas. MMP14 subcellular location (ICC-IF): Mainly localized to the cytosol. In addition localized to the intermediate filaments..
- Human Protein Atlas. MMP14 antibody validation summary (2 antibodies).
- MMP14 expression levels accurately predict the presence of extranodal extensions in oral squamous cell carcinoma: a retrospective cohort study. BMC cancer 2023 — PMC9921360.
- High MMP14 and low decorin expression correlate with disease progression and metastasis in Brazilian prostate cancer patients. Journal of molecular histology 2026 — PMC13303449.
- Global gene expression profiling of human pleural mesotheliomas: identification of matrix metalloproteinase 14 (MMP-14) as potential tumour target. PloS one 2009 — PMC2737627.
- Early intermittent low-dose sclerostin antibody treatment promotes surface bone formation and reduces bone loss, but also decreases osteocyte apoptosis and mechanotransduction in ovariectomized rats: A pilot study. Bone reports 2026 — PMC13101557.
- PubMed PMID:8015608 — UniProt-cited evidence.
- PubMed PMID:7721107 — UniProt-cited evidence.
- PubMed PMID:7708715 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for MMP14 (Matrix metalloproteinase-14). Expected localisation Cytoplasmic staining in tissues (HPA tissue IHC). Antigen retrieval: Citrate pH 6 HIER, heat-mediated (datasheet A00656). Validated IHC antibody: A00656 at 1:25 (the product's own IHC validation image is shown). Controls (HPA): positive Adipose tissue (Adipocytes); negative Bone marrow. IHC-P workflow on paraffin sections. Protocol steps: 1. Sample: Formaldehyde-fixed, paraffin-embedded human lung adenocarcinoma tissue (datasheet A00656); 2. Fixation: Image formalin-fixed; duration unreported (datasheet A00656); 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: Citrate pH 6 (datasheet A00656); 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-MMP14, 1:25 (datasheet A00656); 9. Primary incubation: 1 hours at 37°C (datasheet A00656). Watch for: Fixation: Formaldehyde-fixed paraffin sections are documented (selected-SKU IHC image A00656); Caveat: Staining has medium consistency with RNA expression (HPA tissue IHC); Isoform / epitope: 0 isoforms annotated; extracellular versus cytoplasmic epitope matters (UniProt); Regulation: Specific expression regulation is not reported (UniProt). Evidence: UniProt P50281, HPA tissue IHC · Approved, HPA ICC-IF · Mainly localized to the cytosol. In addition localized to the…, Datasheet A00656, PMC2737627, PMC13101557. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/MMP14/immunohistochemistry-ihc-guide.