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CLTC / Clathrin heavy chain 1 · IHC design guide
Design Immunohistochemistry for CLTC
Plan CLTC IHC on paraffin sections using the catalog antibody at 1:50 (datasheet M03134) and assess cytoplasmic and membranous staining (HPA tissue IHC). Parathyroid glandular cells and placental trophoblasts provide high-staining references (HPA tissue IHC); compare cell types using a matched negative control (standard IHC practice).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
CLTC 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 | General cytoplasmic/membranous staining; parathyroid glandular cells high (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet M03134) | |
| Positive control | Parathyroid gland+4 more · see all | |
| Negative control | Adipose tissue+3 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image M03134) | |
| Caveat | Broad expression does not mean every cell stains; adipocytes are negative (HPA tissue IHC) | |
| Regulation | No expression regulator annotated (UniProt) | |
| Isoform / epitope | 2 isoforms; no extracellular domain; epitope variation unknown (UniProt) |
Section 1
Recommended CLTC IHC & IF Protocols
The catalog antibody has an IHC-P protocol (datasheet M03134). The published option below describes chromogenic CLTC staining in clinical breast cancer tissues (PMC4284514).
| Sample | Paraffin-embedded human liver cancer tissue; fixative not specified (datasheet M03134) |
| Fixation | Image fixative and duration unreported (datasheet M03134); 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 M03134); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet M03134) |
| Primary antibody | Rabbit monoclonal (clone ABDH-3) anti-CLTC, 1:50 (datasheet M03134) |
| Primary incubation | Overnight at 4 °C (datasheet M03134) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet M03134) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | CLTC-positive staining in glandular cells of parathyroid gland (HPA tissue IHC: High). HPA tissue profile: General cytoplasmic and membranous expression. No signal in the no-primary control. |
💡Decision noteStart with EDTA pH 8.0 heat retrieval (datasheet M03134). The published tissue protocol used 10 mM citrate, pH 6.0, at 100°C for 1 h (PMC4284514).
Section 2
What Is the Expected CLTC Staining Pattern?
CLTC should show cytoplasmic and membranous staining, especially in parathyroid glandular cells and placental trophoblasts, where HPA reports high staining (HPA: tissue IHC). CLTC sits on the cytoplasmic face of coated pits and vesicles and has no transmembrane segment (UniProt Q00610: localization, topology). The HPA tissue profile is Approved, with medium consistency between staining and RNA expression (HPA: reliability).
| Cytoplasmic and membranous signal in parathyroid glandular cells or placental trophoblasts. | This matches the reported high staining in these cell populations (HPA: tissue IHC). Score the signal in the named cells and its compartment, rather than treating an entire section as uniformly positive. A cytoplasmic or membrane-associated pattern is consistent with CLTC on the cytoplasmic face of coated pits and vesicles (UniProt Q00610: localization). |
| Moderate cytoplasmic or membranous staining in other listed positive cells. | Medium staining is reported in adrenal, appendix and breast glandular cells; bronchial respiratory epithelial cells; bone marrow hematopoietic cells; and caudate neuronal cells (HPA: tissue IHC). These can support interpretation, but their reported intensity is below that of the high-staining parathyroid and placental populations (HPA: tissue IHC). Compare like cell populations when scoring. |
| Predominantly nuclear staining, or another compartment that dominates the expected pattern. | Reassess specificity and slide quality before calling this CLTC: the tissue profile is generally cytoplasmic and membranous (HPA: tissue IHC). A spindle-associated signal in a mitotic cell is a possible exception because CLTC is also reported at mitotic spindle bridges (UniProt Q00610: localization). Nuclear dominance alone lacks support in the supplied localization records. |
| Strong signal in adipocytes, oral squamous epithelial cells, or muscle cells reported as unstained. | HPA reports no detection in adipocytes, oral mucosal squamous epithelial cells, skeletal myocytes and smooth muscle cells (HPA: tissue IHC). Unexpected staining there raises concern for cross-reactivity or endogenous detection activity (general IHC practice). Treat these as comparison populations, not absolute biological knockouts; the HPA tissue profile has medium staining–RNA consistency (HPA: reliability). |
| Diffuse staining across cells and surrounding tissue, or no signal in a known-positive population. | A widespread haze obscures the reported cell and compartment pattern (HPA: tissue IHC). No signal in parathyroid glandular cells or placental trophoblasts conflicts with their reported high staining (HPA: tissue IHC). Neither result alone identifies a CLTC-specific cause; assess controls, tissue preservation and the general IHC workflow before interpreting a biological difference (general IHC practice). |
💡Expected CLTC appearanceCall a positive result when parathyroid glandular cells or placental trophoblasts show high cytoplasmic and membranous staining (HPA: tissue IHC); nuclear dominance or strong staining in HPA-undetected cell populations warrants investigation before a positive call (HPA: tissue IHC; UniProt Q00610: localization).
| Where should the signal sit? | CLTC is associated with the cytoplasmic face of coated pits and vesicles and has no transmembrane segment (UniProt Q00610: localization, topology). Interpret a membrane-associated rim together with cytoplasmic staining; the supplied record does not support calling CLTC a transmembrane surface marker. |
| How much confidence does tissue validation provide? | The HPA tissue pattern is Approved, with medium consistency between antibody staining and RNA expression (HPA: reliability). The listed HPA antibodies have Approved IHC status (HPA: antibody validation). These ratings support comparison with the reported pattern, while leaving unexpected staining to be checked against controls. |
| Could isoforms change the expected IHC result? | UniProt lists 2 CLTC isoforms (UniProt Q00610: isoforms). The supplied record gives no antibody epitope or isoform-specific tissue pattern, so it cannot predict which isoform a particular IHC signal represents. Avoid assigning regional staining differences to isoforms from this evidence alone. |
| What about IF/ICC localization? | HPA reports enhanced endosome and lysosome localization in ICC-IF, with supported mitotic spindle localization; cytosol and centriolar satellite assignments are uncertain (HPA: subcellular). Those cellular images can inform compartment interpretation, but they do not set an IHC-P staining threshold or provide an IHC protocol. |
| Situation | Likely cause | Next action |
|---|---|---|
| No signal in parathyroid glandular cells or placental trophoblasts. | This conflicts with their reported high staining (HPA: tissue IHC); the cause is unresolved. | Check the positive-control section, antibody dilution, antigen-retrieval record and detection steps (general IHC practice). |
| Only weak staining in a section expected to be strongly positive. | High HPA staining is cell-specific, so a section with few relevant cells can appear weak overall (HPA: tissue IHC). | Locate and score the named glandular or trophoblastic cells; compare a suitable positive-control section (general IHC practice). |
| Strong staining in adipocytes or HPA-undetected epithelial or muscle cells. | Cross-reactivity or endogenous detection activity is possible (HPA: tissue IHC; general IHC practice). | Review a no-primary control and the relevant endogenous-activity block before assigning CLTC positivity (general IHC practice). |
| Diffuse chromogen obscures cell boundaries. | Nonspecific background can mask the reported cytoplasmic and membranous profile (HPA: tissue IHC; general IHC practice). | Compare no-primary control staining; review blocking, washes, antibody dilution and detection time (general IHC practice). |
| Signal appears predominantly nuclear. | Nuclear dominance is unsupported by the supplied tissue and localization profiles (HPA: tissue IHC; UniProt Q00610: localization). | Check counterstain and controls, then reassess whether cytoplasmic or membrane-associated signal is present (general IHC practice). |
| A spindle-like signal appears in a dividing cell. | CLTC can localize to spindle inter-microtubule bridges (UniProt Q00610: localization). | Assess the dividing cell in context and compare the broader cytoplasmic and membranous tissue pattern before scoring (HPA: tissue IHC). |
Experiment evidence
IHC & IF images showing CLTC staining
M03134 · IHCIHC analysis of CLTC using anti-CLTC antibody (M03134). CLTC was detected in a paraffin-embedded section of human liver cancer tissue. Heat
M03134 · IHCIHC analysis of CLTC using anti-CLTC antibody (M03134). CLTC was detected in a paraffin-embedded section of human liver cancer tissue. Heat
M03134 · IFImmunofluorescent analysis using the Antibody at 1:50 dilution.
A03134-1 · IHCIHC analysis of Clathrin heavy chain/CLTC using anti-Clathrin heavy chain/CLTC antibody (A03134-1). Clathrin heavy chain/CLTC was detected i
A03134-1 · IHCIHC analysis of Clathrin heavy chain/CLTC using anti-Clathrin heavy chain/CLTC antibody (A03134-1). Clathrin heavy chain/CLTC was detected i
A03134-1 · IFIF analysis of Clathrin heavy chain/CLTC using anti-Clathrin heavy chain/CLTC antibody (A03134-1). Clathrin heavy chain/CLTC was detected in
M03134-2 · IHCIHC analysis of Clathrin heavy chain/CLTC using anti-Clathrin heavy chain/CLTC antibody (M03134-2). Clathrin heavy chain/CLTC was detected i
M03134-2 · IHCIHC analysis of Clathrin heavy chain/CLTC using anti-Clathrin heavy chain/CLTC antibody (M03134-2). Clathrin heavy chain/CLTC was detected i
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000141367-CLTC/tissue/cerebral+cortex#imid_4428912
HPA · Parathyroid glandHuman Protein Atlas IHC, Parathyroid gland. Source: https://www.proteinatlas.org/ENSG00000141367-CLTC/tissue/parathyroid+gland#imid_4428803
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000141367-CLTC/tissue/colon#imid_4428838
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000141367-CLTC/tissue/liver#imid_4428847
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody CAB011571; green = CLTC, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody CAB011571; green = CLTC, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for CLTC IHC & IF
🧪Run parathyroid gland first; its glandular cells should stain (HPA: High in parathyroid glandular cells). Use skeletal muscle myocytes as the negative tissue (HPA: Not detected in skeletal muscle myocytes); unstained non-glandular cells on the parathyroid slide can indicate background, but HPA does not establish that they are CLTC-negative.
Positive control tissue: Parathyroid gland (Glandular cells, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show CLTC in A-431, U-251MG, U2OS, RPTEC/TERT1, hTERT-RPE1 (serum starved), Sperm, HeLa BAC 4422, HeLa , with annotated localisation: Endosomes (enhanced), Lysosomes (enhanced) (HPA subcellular).
Technical controls: Include no-primary (secondary-only), host- and antibody-format-matched rabbit IgG, and CLTC-knockout tissue or a peptide-block control if the immunizing peptide is available (selected-SKU caption: rabbit anti-CLTC). For chromogenic detection, block endogenous peroxidase and check the parathyroid section for background before scoring (selected-SKU caption: peroxidase/DAB detection).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected-SKU paraffin-section caption does not state the fixative (selected-SKU caption: fixative not stated). Heat retrieval in EDTA at pH 8.0 is a documented starting condition, but retrieval dependence is unreported (selected-SKU caption: heat-mediated EDTA retrieval). The supplied evidence does not establish that frozen sections or IF are easier, and reports no parathyroid-specific artefact; ICC-IF images are available for listed cell lines (HPA: subcellular ICC-IF images).
HPA tissue IHC evidence for CLTC
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 |
|---|---|---|---|---|
| Parathyroid gland | Glandular cells | High | Protein (IHC) | HPA → |
| Placenta | Trophoblastic cells | High | Protein (IHC) | HPA → |
| Adrenal gland | Glandular cells | Medium | Protein (IHC) | HPA → |
| Appendix | Glandular cells | Medium | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Oral mucosa | Squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
| Skeletal muscle | Myocytes | Not detected | Protein (IHC) | HPA → |
| Smooth muscle | Smooth muscle cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced CLTC IHC Tips
Troubleshoot CLTC staining in paraffin sections by checking retrieval, controls, and the expected cytoplasmic and membranous pattern (datasheet M03134; HPA tissue IHC).
How should I retrieve CLTC in paraffin sections with weak staining?
Use heat-mediated retrieval in EDTA buffer at pH 8.0 for this page’s IHC-validated antibody (datasheet M03134). The product image used a paraffin section of human liver cancer tissue, followed by 1:50 primary antibody overnight at 4°C (datasheet M03134). If staining is weak, check that sections stayed covered by retrieval buffer during heating, then compare retrieval duration on adjacent sections while keeping detection conditions constant (standard IHC practice). Include a positive control section, such as placenta with high trophoblastic-cell staining, and assess whether cytoplasmic or membranous signal improves without stronger diffuse background (HPA tissue IHC; standard IHC practice).
Could fixation explain weak or uneven CLTC staining?
Target-specific CLTC sensitivity to fixation is unknown from the supplied evidence; the selected paraffin-section caption does not state a fixative (datasheet M03134). Record the specimen’s fixative, fixation duration, processing history, and section age before changing the antibody protocol, because these variables can affect antigen preservation and accessibility (standard IHC practice). Compare similarly processed positive control sections while retaining EDTA retrieval at pH 8.0 and the documented 1:50 primary dilution (datasheet M03134; standard IHC practice). Treat staining differences between differently processed specimens as a possible technical effect until matched controls support a biological interpretation (standard IHC practice).
What CLTC staining pattern should I accept as plausible?
Expect predominantly cytoplasmic and membranous staining in tissue sections, with possible punctate signal reflecting coated pits and cytoplasmic vesicles (HPA tissue IHC; UniProt Q00610 subcellular location). CLTC faces the cytoplasm on coated pits and vesicles and has no transmembrane segment, so a purely nuclear pattern needs independent validation (UniProt Q00610 topology and subcellular location). In dividing cells, spindle-associated staining can be plausible through the TACC3/ch-TOG/clathrin complex, but assess cell morphology before assigning that pattern (UniProt Q00610 subunit and function; standard IHC practice). Compare the candidate signal with adjacent cell populations and a no-primary control to distinguish structured staining from chromogen deposits (standard IHC practice).
Can this antibody distinguish CLTC isoforms or modified epitopes in IHC?
CLTC has 2 annotated isoforms, but the supplied product caption does not identify the antibody epitope or establish isoform selectivity (UniProt Q00610 isoforms; datasheet M03134). The protein also carries annotated modified residues, including phosphorylation at positions 67, 105, 184, and 394; their effects on this antibody’s staining are unknown (UniProt Q00610 modified residues; datasheet M03134). Do not interpret stronger DAB staining as a particular isoform or modification state without epitope and specificity evidence (standard IHC practice). If that distinction matters, obtain the antibody’s immunogen or epitope information and validate against an independently defined isoform or modification control (standard IHC practice).
How should I adapt CLTC localisation checks to multiplex IF?
For a separate IF experiment, pair CLTC with a marker identifying the expected cell type so puncta can be assigned to the correct cells (standard IF practice). Choose fluorophores and imaging channels after checking tissue autofluorescence, and include single-label controls to assess spectral bleed-through (standard IF practice). CLTC lies on the cytoplasmic face of coated pits and vesicles, so test permeabilisation sufficient to expose that side while preserving punctate structure (UniProt Q00610 topology and subcellular location; standard IF practice). Endosomal and lysosomal localisation is reported in cell imaging, but assess its relevance to the specimen and antibody used before interpreting colocalisation (HPA subcellular; standard IF practice).
How can I reduce diffuse DAB or staining outside expected cells?
The documented IHC example used 10% goat serum blocking, 1:50 primary antibody overnight at 4°C, and an HRP/DAB detection workflow (datasheet M03134). If background obscures cytoplasmic or membranous CLTC signal, compare a no-primary control and check wash quality, blocking, and antibody concentration on adjacent sections (HPA tissue IHC; standard IHC practice). Include a peroxidase-blocking step and inspect the no-primary section for residual endogenous enzyme activity; these are general chromogenic IHC controls, not CLTC-specific validation (standard IHC practice). Shorten chromogen development if nonspecific deposits increase while the positive control retains interpretable cellular staining (standard IHC practice).
How should I score CLTC staining across specimens? ⚠ ANSWER MARKED FOR VERIFICATION
Define the scored cell population and compartment before measuring CLTC, because tissue IHC shows general cytoplasmic and membranous expression (HPA tissue IHC; standard IHC practice). For intensity-based comparison, record the percentage of cells at each staining intensity and calculate an H-score using the same thresholds across specimens (standard IHC practice). Alternatively, report the percentage of positive cells or positive-cell density per mm² when intensity is unreliable, and normalise counts to the evaluated cell population or viable tissue area (standard IHC practice). Keep section processing, retrieval at pH 8.0, image acquisition, and scoring rules consistent, with controls on each staining run (datasheet M03134; standard IHC practice).
How do I distinguish genuine CLTC staining from artefact?
A credible pattern should align with cytoplasmic and membranous tissue staining, with puncta compatible with coated pits or vesicles (HPA tissue IHC; UniProt Q00610 subcellular location). Check the stained cell type: HPA reports high signal in placental trophoblastic and parathyroid glandular cells, while adipocytes and skeletal myocytes were not detected in its tissue profiles (HPA tissue IHC). Treat isolated nuclear staining, section-edge enhancement, necrotic deposits, or signal persisting without primary antibody as reasons to investigate artefact (UniProt Q00610 topology; standard IHC practice). Review morphology, a positive control, and the no-primary control, including residual endogenous peroxidase staining, before calling a specimen CLTC-positive (standard IHC practice).
Boster reagents
Best CLTC / Clathrin heavy chain 1 IHC Antibodies
Catalog images show CLTC staining in human paraffin sections, mouse and rat intestine sections, and U20S cells by IF/ICC (catalog image captions: M03134, A03134-1, M03134-2).
Real IHC data
Anti-Clathrin heavy chain Rabbit Monoclonal Antibody
Cat # M03134
Real IHC data
Anti-Clathrin heavy chain/CLTC Antibody ®
Cat # A03134-1
Real IHC data
Anti-Clathrin heavy chain/CLTC Antibody ® (monoclonal, 6D3)
Cat # M03134-2
M03134 shows IHC staining in human liver and lung cancer paraffin sections; A03134-1 shows IHC staining in human lung cancer and mouse and rat intestine paraffin sections, plus IF/ICC staining in U20S cells (catalog image captions: M03134, A03134-1). M03134-2 shows IHC staining in human pancreatic cancer, renal carcinoma, and tonsil paraffin sections (catalog image captions: M03134-2).
Which to pick: For tissue IHC, choose rabbit monoclonal M03134 when its human liver or lung cancer paraffin examples match the planned sample; its caption uses EDTA retrieval and 1:50 primary antibody (catalog entry and IHC image caption: M03134). For IF/ICC, choose A03134-1 because its application list includes both and its cell image documents staining at 5 μg/mL (catalog application list and IF image caption: A03134-1). For IHC across human, mouse, and rat samples, choose A03134-1 because it has a paraffin section image for each species; the cited tissue captions do not report the fixative (catalog IHC image captions: A03134-1).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q00610 (CLH1_HUMAN, Clathrin heavy chain 1).
- Human Protein Atlas. CLTC tissue IHC expression (reliability: Approved).
- Human Protein Atlas. CLTC subcellular location (ICC-IF): Mainly localized to the endosomes and lysosomes. In addition localized to the cytosol, centriolar satellites, mitotic spindle and mid piece..
- Human Protein Atlas. CLTC antibody validation summary (4 antibodies).
- MicroRNA-574-3p, identified by microRNA library-based functional screening, modulates tamoxifen response in breast cancer. Scientific reports 2015 — PMC4284514.
- Enamel defects in Acp4(R110C/R110C) mice and human ACP4 mutations. Scientific reports 2022 — PMC9526733.
- Epithelioid Inflammatory Myofibroblastic Sarcoma: Case Series With a First Report of CLTC::ALK Fusion in an Aggressive Disease. Genes, chromosomes & cancer 2025 — PMC12086959.
- PubMed PMID:7584026 — UniProt-cited evidence.
- PubMed PMID:17974005 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for CLTC (Clathrin heavy chain 1). Expected localisation Cytoplasmic and membranous tissue staining (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet M03134). Validated IHC antibody: M03134 at 1:50 (the product's own IHC validation image is shown). Controls (HPA): positive Parathyroid gland (Glandular cells); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human liver cancer tissue; fixative not specified (datasheet M03134); 2. Fixation: Image fixative and duration unreported (datasheet M03134); 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 M03134); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet M03134); 8. Primary antibody: Rabbit monoclonal (clone ABDH-3) anti-CLTC, 1:50 (datasheet M03134); 9. Primary incubation: Overnight at 4 °C (datasheet M03134). Watch for: Fixation: Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image M03134); Caveat: Broad expression does not mean every cell stains; adipocytes are negative (HPA tissue IHC); Isoform / epitope: 2 isoforms; no extracellular domain; epitope variation unknown (UniProt); Regulation: No expression regulator annotated (UniProt). Evidence: UniProt Q00610, HPA tissue IHC · Approved, HPA ICC-IF · Mainly localized to the endosomes and lysosomes. In addition…, Datasheet M03134, PMC4284514. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/CLTC/immunohistochemistry-ihc-guide.