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PDLIM7 / PDZ and LIM domain protein 7 · IHC design guide
Design Immunohistochemistry for PDLIM7
Plan chromogenic PDLIM7 IHC in paraffin sections using cytoplasmic muscle staining as a reference (HPA tissue IHC). The guide covers fixation, controls and scoring, with 2–5 μg/ml as the catalog antibody’s starting concentration (datasheet A04377-1).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
PDLIM7 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 | Cytoplasm (HPA tissue IHC); cytoskeleton (UniProt) | |
| Staining pattern | Cytoplasmic staining in myocytes and cardiomyocytes (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A04377-1) | |
| Positive control | Heart muscle+1 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 | Low consistency between staining and RNA expression (HPA tissue IHC) | |
| Regulation | Most abundant in skeletal muscle (HPA tissue IHC) | |
| Isoform / epitope | Six isoforms; epitope coverage is unknown (UniProt) |
Section 1
Recommended PDLIM7 IHC & IF Protocols
The catalog antibody’s IHC-P protocol uses EDTA pH 8.0 heat retrieval (datasheet A04377-1). One published PDLIM7 IHC protocol provides additional staining parameters (PMC12274469).
| Sample | Paraffin-embedded human appendiceal adenocarcinoma tissue; fixative not specified (datasheet A04377-1) |
| Fixation | Image fixative and duration unreported (datasheet A04377-1); 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 A04377-1); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A04377-1) |
| Primary antibody | Rabbit anti-PDLIM7, 2-5 μg/ml (datasheet A04377-1) |
| Primary incubation | Overnight at 4 °C (datasheet A04377-1) |
| Detection | Streptavidin-biotin complex (SABC), DAB chromogen (datasheet A04377-1) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | PDLIM7-positive staining in cardiomyocytes of heart muscle (HPA tissue IHC: Medium). HPA tissue profile: Cytoplasmic expression in several tissues, most abundant in skeletal muscle. No signal in the no-primary control. |
💡Decision noteStart with EDTA pH 8.0 heat retrieval for the catalog antibody (datasheet A04377-1); the published protocol does not specify its retrieval conditions (PMC12274469 methods).
Section 2
What Is the Expected PDLIM7 Staining Pattern?
PDLIM7 is a cytoplasmic, cytoskeleton-associated protein with no transmembrane segment (UniProt Q9NR12). In paraffin-section IHC, expect cytoplasmic staining in skeletal myocytes and cardiomyocytes, both scored Medium by HPA (HPA tissue IHC). HPA describes cytoplasmic expression across several tissues, most abundant in skeletal muscle, but rates its tissue IHC evidence Approved with low consistency between staining and RNA expression (HPA tissue IHC).
| Cytoplasmic chromogen in skeletal myocytes or cardiomyocytes at roughly Medium intensity (HPA tissue IHC). | This matches HPA's observed positive cells and UniProt's cytoplasmic, cytoskeletal localisation (HPA tissue IHC; UniProt Q9NR12). Assess staining in the identified cells, since a tissue section contains more than one cell type. |
| Predominantly nuclear staining or a continuous cell-surface outline, without convincing cytoplasmic staining. | Treat this as a localisation mismatch: UniProt places PDLIM7 in the cytoplasm and cytoskeleton and lists no transmembrane segment (UniProt Q9NR12). Review antibody specificity and detection controls before scoring it as positive. |
| Strong staining in adipocytes or appendix glandular cells, where HPA reports Not detected (HPA tissue IHC). | This raises concern for cross-reactivity or endogenous detection activity; check a no-primary control and the detection reagents. HPA's negative calls describe those cell populations, not every cell in the section (HPA tissue IHC). |
| Widespread, poorly defined chromogen obscures cell boundaries and the expected cytoplasmic pattern. | A diffuse field cannot establish PDLIM7 localisation. General IHC practice is to inspect no-primary controls, wash quality, blocking and chromogen development before interpreting cell-specific staining. |
| No staining in skeletal myocytes or cardiomyocytes, the HPA Medium positive populations (HPA tissue IHC). | Consider an assay failure before calling the specimen negative. Check section integrity and the IHC-validated antibody's stated procedure, then compare a known-positive section processed in the same run (HPA tissue IHC). |
💡Expected PDLIM7 appearanceCall a section positive when identifiable skeletal myocytes or cardiomyocytes show cytoplasmic staining around Medium intensity; isolated nuclear, sharp membrane-only or control-associated diffuse staining is suspect (HPA tissue IHC; UniProt Q9NR12).
| Cell compartment and topology | UniProt places PDLIM7 in the cytoplasm and cytoskeleton and reports no transmembrane segment (UniProt Q9NR12). Judge paraffin IHC by cellular localisation; a membrane-only outline does not match that annotation. |
| Tissue and cell selection | Skeletal myocytes and cardiomyocytes are Medium IHC positives; adipocytes and appendix glandular cells are Not detected in HPA's observations (HPA tissue IHC). Select controls by cell type. |
| Isoforms and antibody coverage | UniProt lists 6 isoforms and reports isoform 2 predominating in skeletal muscle (UniProt Q9NR12). The supplied evidence gives no antibody epitope, so it cannot establish which isoforms an IHC stain detects. |
| Strength of IHC evidence | HPA rates tissue IHC Approved but notes low staining–RNA consistency; antibody HPA018794 is IHC Approved, not IHC Enhanced (HPA tissue IHC; HPA antibodies). Interpret unexpected positives cautiously. |
| IF/ICC Q: What localisation should I expect? | A: HPA ICC-IF places PDLIM7 mainly on actin filaments and focal adhesion sites; images are listed for A-431, U-251MG and U2OS (HPA subcellular). Those ICC-IF observations do not specify a paraffin IHC protocol. |
| Situation | Likely cause | Next action |
|---|---|---|
| The known-positive muscle section has no chromogen. | The run may have failed; HPA reports Medium staining in skeletal myocytes and cardiomyocytes (HPA tissue IHC). | Check antibody dilution and detection against the IHC-validated antibody's instructions; inspect section integrity and repeat with a positive control in the same run. |
| Signal is mainly nuclear or forms a crisp membrane ring. | That pattern conflicts with cytoplasmic and cytoskeletal localisation and the lack of a transmembrane segment (UniProt Q9NR12). | Compare the no-primary control, examine cell boundaries with the counterstain and reassess specificity before assigning a positive score. |
| Adipocytes or appendix glandular cells stain strongly. | Those cells are Not detected in HPA tissue IHC; cross-reactivity or endogenous detection activity is possible (HPA tissue IHC). | Run a no-primary control and check endogenous enzyme blocking where applicable; score the named cell population separately from neighboring cells. |
| The section has diffuse brown haze. | General IHC practice: background can arise from detection reagents, insufficient washing or overdevelopment. | Inspect a no-primary control, review blocking and washing, and shorten chromogen development if the control shows background. |
| A low or negative cell population differs from the HPA image. | HPA's tissue IHC has low consistency with RNA data, and its staining levels describe observed samples (HPA tissue IHC). | Confirm cell identity, compare same-run positive and negative controls, and describe the observed pattern without treating HPA levels as absolute thresholds. |
| Staining varies markedly across one section. | General IHC practice: uneven reagent coverage or section quality can create spatial variation. | Inspect tissue preservation and reagent coverage; repeat the section with uniform incubation and use the same-run muscle control to assess the run. |
Experiment evidence
IHC & IF images showing PDLIM7 staining
A04377-1 · IHCIHC analysis of PDLIM7 using anti-PDLIM7 antibody (A04377-1). PDLIM7 was detected in a paraffin-embedded section of human appendiceal adenoc
A04377-1 · IHCIHC analysis of PDLIM7 using anti-PDLIM7 antibody (A04377-1). PDLIM7 was detected in a paraffin-embedded section of human gall bladder adeno
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000196923-PDLIM7/tissue/cerebral+cortex#imid_5154396
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000196923-PDLIM7/tissue/colon#imid_5154338
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000196923-PDLIM7/tissue/liver#imid_5154331
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000196923-PDLIM7/tissue/kidney#imid_5154332
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA018794; green = PDLIM7, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA018794; green = PDLIM7, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for PDLIM7 IHC & IF
🧪Run heart muscle first and look for staining in cardiomyocytes (HPA: Medium in cardiomyocytes). Use adipose tissue adipocytes as the negative tissue (HPA: Not detected in adipocytes); on the heart slide, adjacent noncardiomyocyte cells should lack the cardiomyocyte staining pattern, but they are not a validated target-negative population (UniProt Q9NR12: isoforms 1 and 2 are expressed ubiquitously).
Positive control tissue: Heart muscle (Cardiomyocytes, HPA Medium)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show PDLIM7 in A-431, U-251MG, U2OS, with annotated localisation: Actin filaments (enhanced), Focal adhesion sites (enhanced) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control and a concentration-matched rabbit IgG isotype control for the captioned primary antibody; use a PDLIM7 knockout sample or cognate-peptide competition, if available, as a biological specificity control (caption: rabbit anti-PDLIM7 antibody). Quench endogenous peroxidase and block endogenous biotin when using the captioned biotin–SABC/DAB detection in heart muscle (caption: biotinylated secondary, SABC and DAB; standard IHC practice).
⚠️Feasibility: A target-specific fixation window and fixation effect are unreported, and the selected A04377-1 caption does not state the fixative (caption: fixative unreported). The paraffin-section example used heat retrieval in EDTA at pH 8.0, but whether retrieval is required has not been established by a matched comparison (caption: EDTA heat retrieval). IF may help assess the reported actin-filament and focal-adhesion localization, but the supplied evidence does not establish that IF or frozen sections are easier; heart-muscle peroxidase background needs control during chromogenic IHC (HPA: actin filaments and focal adhesion sites; standard IHC practice).
HPA tissue IHC evidence for PDLIM7
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 |
|---|---|---|---|---|
| Heart muscle | Cardiomyocytes | Medium | Protein (IHC) | HPA → |
| Skeletal muscle | Myocytes | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Appendix | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic 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 PDLIM7 IHC Tips
Troubleshoot PDLIM7 chromogenic IHC in paraffin sections with attention to retrieval, cytoplasmic localisation, and cell specific controls (caption A04377-1; HPA tissue IHC).
Which retrieval conditions should I start with for PDLIM7 in paraffin sections?
Start with heat mediated retrieval in EDTA at pH 8.0 for paraffin sections (datasheet A04377-1). The selected image used that treatment before overnight incubation at 4°C with 2 μg/ml primary antibody, so keep those conditions together for the first comparison (caption A04377-1). If signal is weak, compare a controlled change in heating duration or a citrate buffer fallback while holding section thickness, detection, and exposure to DAB constant (standard IHC practice). Judge improvement by specific cytoplasmic staining in myocytes alongside minimal signal in a negative control, since HPA reports myocyte positivity but low staining to RNA consistency (HPA tissue IHC).
How should I troubleshoot fixation when PDLIM7 staining is weak or uneven?
The selected paraffin section caption does not report a fixative, so target specific fixation sensitivity is unknown (caption A04377-1). Record fixative, fixation duration, processing history, and section age for each specimen before attributing weak staining to fixation (standard IHC practice). Process a small matched series consistently, then compare EDTA retrieval at pH 8.0 and 2 μg/ml primary antibody across the series (datasheet A04377-1; caption A04377-1). Assess intact cytoplasmic staining and tissue morphology together, using myocytes as a positive reference where available (HPA tissue IHC), and treat uneven peripheral staining or damaged areas as possible processing artefacts (standard IHC practice).
What PDLIM7 staining pattern should I expect in chromogenic tissue sections?
Expect predominantly cytoplasmic staining in tissue IHC, with particularly abundant expression in skeletal muscle (HPA tissue IHC). UniProt also places PDLIM7 in the cytoplasm and cytoskeleton and describes colocalisation near the muscle Z line, which makes structured myocyte staining biologically plausible (UniProt Q9NR12 subcellular). The protein has no transmembrane segment, so an isolated continuous cell surface rim needs independent validation before it is scored as PDLIM7 (UniProt Q9NR12 topology). Compare myocytes and cardiomyocytes, both reported at medium staining, with local negative controls and morphology rather than assuming every brown deposit marks the intended compartment (HPA tissue IHC; standard IHC practice).
Could PDLIM7 isoforms or epitope masking explain discordant tissue staining?
PDLIM7 has 6 annotated isoforms, but the supplied antibody caption does not identify its epitope or establish isoform coverage (UniProt Q9NR12 isoforms; caption A04377-1). Isoforms 1 and 2 are ubiquitous, with isoform 2 predominant in skeletal muscle and isoform 1 more abundant in lung, spleen, leukocytes, and fetal liver (UniProt Q9NR12 tissue specificity). Map the immunogen against isoform sequences before claiming that differing IHC intensity reflects expression rather than recognition (standard antibody validation practice). The PDZ domain spans residues 1–85, and three LIM domains occupy residues 280–457; retrieval dependent masking remains a hypothesis until tested with a mapped epitope and matched sections (UniProt Q9NR12 domains; standard IHC practice).
How can IF help validate PDLIM7 localisation seen by chromogenic IHC?
Use IF to check localisation, pairing PDLIM7 with a marker for the expected cell type and an actin or focal adhesion marker (HPA subcellular; standard IF practice). HPA reports enhanced localisation at actin filaments and focal adhesion sites in ICC/IF, whereas tissue IHC describes cytoplasmic staining (HPA subcellular; HPA tissue IHC). Choose fluorophores in spectral regions with low autofluorescence and include unstained and single stain controls (standard IF practice). Because PDLIM7 lacks a transmembrane segment and is cytoplasmic, permeabilise sufficiently to access its intracellular epitope, then check that this treatment preserves the structures being compared (UniProt Q9NR12 topology; UniProt Q9NR12 subcellular; standard IF practice).
How do I distinguish PDLIM7 signal from nonspecific DAB background?
The selected paraffin image used 10% goat serum blocking, 2 μg/ml primary overnight at 4°C, and a biotinylated secondary followed by a streptavidin biotin complex with DAB (caption A04377-1). First compare no primary and secondary only controls, and verify that endogenous peroxidase has been blocked before interpreting brown deposits (standard chromogenic IHC practice). If background remains, titrate the primary downward and shorten DAB development while retaining EDTA retrieval at pH 8.0 during the comparison (datasheet A04377-1; standard IHC practice). Inspect tissue edges, folds, necrotic regions, and pigment separately, then require cytoplasmic staining in plausible cells before assigning specific PDLIM7 signal (HPA tissue IHC; standard IHC practice).
How should I quantify PDLIM7 IHC across tissues with different cell composition? ⚠ ANSWER MARKED FOR VERIFICATION
Segment viable tissue and define the cell population before measuring DAB, because HPA reports medium staining in myocytes and cardiomyocytes with varying results elsewhere (HPA tissue IHC). Report percentage of positive target cells and an H score based on intensity categories, using consistent thresholds across slides (standard IHC quantification practice). For spatial comparisons, report positive cells per mm² of target compartment and normalise to counted target cells or compartment area (standard IHC quantification practice). Keep acquisition, counterstain, and DAB development consistent, and exclude folds or necrosis; interpret small differences cautiously because HPA labels antibody staining versus RNA consistency as low (HPA tissue IHC; standard IHC practice).
What evidence makes a PDLIM7 positive IHC result convincing?
A convincing result shows cellular cytoplasmic staining with myocytes and cardiomyocytes as references at medium intensity (HPA tissue IHC; UniProt Q9NR12 subcellular). Evaluate whether signal follows cell morphology rather than section edges, folds, or necrotic tissue, and check a no primary control (standard IHC practice). A purely nuclear pattern or uninterrupted membrane rim needs validation because reported locations are cytoplasm and cytoskeleton, with no transmembrane segment (UniProt Q9NR12 subcellular; UniProt Q9NR12 topology). The selected appendiceal adenocarcinoma image demonstrates staining after EDTA pH 8.0 retrieval, but appendix glandular cells were not detected in HPA tissue IHC; compare cell identity and controls (caption A04377-1; HPA tissue IHC).
Boster reagents
Best PDLIM7 / PDZ and LIM domain protein 7 IHC Antibodies
The IHC-validated antibody A04377-1 has real staining data from human paraffin-embedded carcinoma sections (catalog image captions). Mouse and Rat reactivity is listed, while IF/ICC validation is unreported (catalog applications and reactivity).
Real IHC data
Anti-PDLIM7 Antibody ®
Cat # A04377-1
A04377-1 has IHC images from human paraffin-embedded appendiceal adenocarcinoma, gall bladder adenosquamous carcinoma, and gastric adenocarcinoma sections (catalog image captions). Its listed applications include IHC, and its listed reactivity is Human, Mouse, and Rat; the images document human IHC only (catalog applications, reactivity, and image captions).
Which to pick: Choose A04377-1 for human paraffin-section IHC: its caption uses 2 μg/ml after EDTA retrieval at pH 8.0, and its listed IHC range is 2–5 μg/ml (A04377-1 image caption; catalog dilution). The caption does not report the fixative, and IF/ICC is absent from the application list, so there is no documented IF/ICC option here (A04377-1 image caption; catalog applications). A04377-1 lists Mouse and Rat reactivity, but the supplied IHC images show human tissue only; cross-species tissue IHC would require validation (catalog reactivity and image captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q9NR12 (PDLI7_HUMAN, PDZ and LIM domain protein 7).
- Human Protein Atlas. PDLIM7 tissue IHC expression (reliability: Approved).
- Human Protein Atlas. PDLIM7 subcellular location (ICC-IF): Mainly localized to the actin filaments and focal adhesion sites..
- Human Protein Atlas. PDLIM7 antibody validation summary (2 antibodies).
- The effect of PDLIM7 on cell proliferation, migration, and drug sensitivity in clear cell renal cell carcinoma. Scientific reports 2025 — PMC12274469.
- TPM2 attenuates progression of prostate cancer by blocking PDLIM7-mediated nuclear translocation of YAP1. Cell & bioscience 2023 — PMC9948342.
- H3K27 acetylation activated-PDLIM7 promotes castration-resistant prostate cancer progression by inducing O-Glycosylation of YAP1 protein. Translational oncology 2024 — PMC10714362.
- PubMed PMID:7929196 — UniProt-cited evidence.
- PubMed PMID:11874232 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for PDLIM7 (PDZ and LIM domain protein 7). Expected localisation Cytoplasm (HPA tissue IHC); cytoskeleton (UniProt). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A04377-1). Validated IHC antibody: A04377-1 at 2-5 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Heart muscle (Cardiomyocytes); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human appendiceal adenocarcinoma tissue; fixative not specified (datasheet A04377-1); 2. Fixation: Image fixative and duration unreported (datasheet A04377-1); 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 A04377-1); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A04377-1); 8. Primary antibody: Rabbit anti-PDLIM7, 2-5 μg/ml (datasheet A04377-1); 9. Primary incubation: Overnight at 4 °C (datasheet A04377-1). Watch for: Fixation: Keep fixation consistent across sections. (standard IHC practice; not target-specific); Caveat: Low consistency between staining and RNA expression (HPA tissue IHC); Isoform / epitope: Six isoforms; epitope coverage is unknown (UniProt); Regulation: Most abundant in skeletal muscle (HPA tissue IHC). Evidence: UniProt Q9NR12, HPA tissue IHC · Approved, HPA ICC-IF · Mainly localized to the actin filaments and focal adhesion sites., Datasheet A04377-1, PMC12274469. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/PDLIM7/immunohistochemistry-ihc-guide.