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CEP70 / Centrosomal protein of 70 kDa · IHC design guide
Design Immunohistochemistry for CEP70
Plan chromogenic CEP70 IHC in paraffin sections using the catalog antibody's 1:100–1:300 dilution range (datasheet A10748-1). Assess cytoplasmic staining across cell types, including kidney tubules with high staining and adipocytes with no detected staining (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
CEP70 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 in tissue sections (HPA tissue IHC) | |
| Staining pattern | Widespread cytoplasmic staining across cell types (HPA tissue IHC) | |
| Antigen retrieval | Tris-EDTA pH 8.0 HIER, heat-mediated (datasheet A10748-1) | |
| Positive control | Caudate+4 more · see all | |
| Negative control | Adipose tissue+1 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep paraffin tissue fixation consistent across samples. (standard IHC practice; not target-specific) | |
| Caveat | Adipocytes and cholangiocytes may lack staining (HPA tissue IHC) | |
| Regulation | Low tissue specificity (HPA tissue RNA) | |
| Isoform / epitope | 5 isoforms; epitope coverage is unspecified (UniProt) |
Section 1
Recommended CEP70 IHC & IF Protocols
The catalog antibody’s IHC-P protocol (datasheet A10748-1) is accompanied by one published pancreatic tissue IHC protocol (PMC4759539).
| Sample | Paraffin-embedded Human brain tissue; fixative not specified (datasheet A10748-1) |
| Fixation | Image fixative and duration unreported (datasheet A10748-1); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat retrieval: Tris-EDTA pH 8.0 (datasheet A10748-1); 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-CEP70, 1:100-1:300 (datasheet A10748-1) |
| Primary incubation | Overnight at 4 °C (standard) |
| Detection | HRP-polymer secondary, DAB chromogen 5–10 min (standard) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | CEP70-positive staining in neuronal cells of caudate (HPA tissue IHC: High). HPA tissue profile: Ubiquitous cytoplasmic expression. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated Tris-EDTA pH 8.0 retrieval (datasheet A10748-1); the article reports antigen retrieval without specifying its conditions (PMC4759539).
Section 2
What Is the Expected CEP70 Staining Pattern?
CEP70 localizes to the cytoplasm and centrosome, including spindle poles during mitosis (UniProt Q8NHQ1). In tissue IHC, expect predominantly cytoplasmic staining across many cell types, with high staining in selected neuronal, tubular, glandular, endothelial and muscle cells (HPA: ubiquitous cytoplasmic expression; High in listed cells). HPA rates tissue IHC reliability Enhanced, with medium consistency between staining and RNA data (HPA: tissue IHC). CEP70 has no transmembrane segment (UniProt Q8NHQ1 topology).
| Cytoplasmic staining in kidney tubular cells, cerebellar Purkinje cells or pancreatic exocrine cells. | This fits the reported tissue pattern: each listed cell population stains High (HPA: tissue IHC). A small centrosomal focus may fit CEP70 localization, but a visible focus is not required to call a chromogenic section positive (UniProt Q8NHQ1 localization; general IHC practice). |
| Strong nuclear-only staining with little cytoplasmic signal in tissue IHC. | Reassess specificity and detection conditions: nuclear-only staining differs from HPA's ubiquitous cytoplasmic tissue profile (HPA: tissue IHC). Do not treat every nuclear signal as false; nucleoplasm is an uncertain ICC-IF location, a separate assay observation (HPA: subcellular ICC-IF). |
| Prominent staining in adipocytes or liver cholangiocytes. | These cells are reported Not detected, so check cross-reactivity or endogenous chromogen activity before assigning the signal to CEP70 (HPA: tissue IHC; general IHC practice). The finding does not make the entire adipose or liver section a CEP70-negative control (HPA: cell-specific tissue IHC entries). |
| Diffuse color across cells and surrounding tissue, obscuring cell boundaries. | A broadly distributed haze cannot establish the reported cellular pattern (HPA: ubiquitous cytoplasmic expression; general IHC practice). Compare a no-primary control and inspect blocking, washing and chromogen development for nonspecific background (general IHC practice). |
| No staining in kidney tubular cells despite intact tissue morphology. | Kidney tubular cells are a reported High population, making absent signal a reason to check assay performance (HPA: tissue IHC). Review the antibody's IHC validation and the run controls before interpreting the sample as biologically negative (HPA: IHC Enhanced for HPA036941 and HPA036942; general IHC practice). |
💡Expected CEP70 appearanceCall a tissue section positive when cytoplasmic staining is convincing in a reported High cell population, such as kidney tubular cells; strong nuclear-only color or staining dominated by background warrants review (HPA: tissue IHC; general IHC practice).
| Cell-specific reference pattern | High staining is reported in neuronal cells of the caudate and cerebral cortex, cerebellar Purkinje cells, kidney tubular cells, pancreatic exocrine cells, placental endothelial cells, rectal glandular cells and skeletal myocytes (HPA: tissue IHC). Compare like cell types when judging intensity (general IHC practice). |
| Validation and limits | HPA036941 and HPA036942 each have Enhanced IHC validation, while the tissue profile has medium consistency with RNA expression data (HPA: antibodies; HPA: tissue IHC). This supports pattern comparison but does not prove that every stained structure is CEP70 (general IHC practice). |
| Subcellular context | CEP70 is at the centrosome in interphase and spindle poles in mitosis (UniProt Q8NHQ1 localization). Tissue IHC is described as ubiquitously cytoplasmic (HPA: tissue IHC); absence of a resolvable centrosomal dot alone should not overturn an otherwise fitting tissue pattern (general IHC practice). |
| Isoforms and antigen coverage | Five isoforms are listed (UniProt Q8NHQ1 isoforms). The supplied record gives no antibody epitope or isoform coverage, so differences between isoforms cannot be used to explain a particular staining result (UniProt Q8NHQ1 isoforms; supplied antibody record). |
| IF/ICC Q&A: What localization is reported? | HPA reports primary cilium as approved, centrosome as supported and basal body as approved; nucleoplasm and cytosol are uncertain ICC-IF locations (HPA: subcellular ICC-IF). Interpret those ratings within IF/ICC rather than requiring the same visible structures in chromogenic tissue IHC (general microscopy practice). |
| Situation | Likely cause | Next action |
|---|---|---|
| Reported High cells show no chromogenic signal. | The run may have failed, or the tested section may differ from the HPA reference; the observation alone cannot distinguish these explanations (HPA: tissue IHC; general IHC practice). | Check a reported High cell population in a control section, then review the primary antibody, retrieval and detection steps against the IHC-validated antibody's instructions (HPA: tissue IHC; general IHC practice). |
| The entire section has a diffuse brown haze. | Nonspecific reagent binding or excess chromogen development can obscure cellular staining (general IHC practice). | Inspect a no-primary control; review blocking, wash stringency and development time, then compare whether the cell-specific cytoplasmic pattern emerges (general IHC practice; HPA: tissue IHC). |
| Signal is strongest in adipocytes or cholangiocytes. | Those cell populations are reported Not detected; cross-reactivity or endogenous detection activity is possible, but the slide alone does not identify the cause (HPA: tissue IHC; general IHC practice). | Check no-primary and detection controls, and compare with a reported High population on a suitable section before assigning CEP70 positivity (general IHC practice; HPA: tissue IHC). |
| Nuclear staining dominates the tissue section. | This differs from the cytoplasmic tissue IHC profile, although nucleoplasm is listed as uncertain in ICC-IF (HPA: tissue IHC; HPA: subcellular ICC-IF). | Assess cytoplasmic signal and controls, then compare the cell-type pattern with HPA tissue IHC; keep the ICC-IF nuclear observation as a caveat rather than proof of tissue IHC specificity (HPA: tissue IHC; HPA: subcellular ICC-IF; general IHC practice). |
| Only isolated puncta are visible, with no convincing cytoplasmic staining. | Centrosomal localization makes a punctum plausible, but morphology alone cannot identify it as CEP70; HPA describes the tissue IHC profile as cytoplasmic (UniProt Q8NHQ1 localization; HPA: tissue IHC). | Check puncta against no-primary controls and the expected cell-type distribution; avoid calling a section positive solely from isolated dots (general IHC practice; HPA: tissue IHC). |
| A low-staining reference cell type is used as a negative control. | Salivary glandular cells, ovarian stromal cells and splenic white-pulp cells are reported Low, which is distinct from Not detected (HPA: tissue IHC). | Label the comparator by its reported cell-specific level, and use an assay control to assess nonspecific color rather than treating Low expression as a guaranteed blank (HPA: tissue IHC; general IHC practice). |
Experiment evidence
IHC & IF images showing CEP70 staining
A10748-1 · IHCImmunohistochemical analysis of paraffin-embedded Human brain. Antibody was diluted at 1:100 (4° overnight). High-pressure and temperature T
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000114107-CEP70/tissue/cerebral+cortex#imid_9392444
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000114107-CEP70/tissue/colon#imid_9387610
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000114107-CEP70/tissue/liver#imid_9387611
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000114107-CEP70/tissue/kidney#imid_9387628
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA036941; green = CEP70, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA036941; green = CEP70, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for CEP70 IHC & IF
🧪Run caudate first: neuronal cells should stain (HPA: High in caudate neuronal cells; catalog antibody caption: paraffin-embedded human brain). Use adipose tissue as the negative tissue; adipocytes should show no specific staining (HPA: Not detected in adipocytes). On the caudate slide, assess non-neuronal cells for background-level staining as provisional internal negatives; their CEP70 status is not established by the supplied HPA row (HPA: High in neuronal cells).
Positive control tissue: Caudate (Neuronal cells, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show CEP70 in A-431, U-251MG, U2OS, ASC52telo, RPTEC/TERT1, hTERT-RPE1 (serum starved), with annotated localisation: Nucleoplasm (uncertain), Primary cilium (approved), Cytosol (uncertain) (HPA subcellular).
Technical controls: Include no-primary (secondary-only) and isotype controls, matching the isotype antibody to the primary antibody’s host species and clonality (standard IHC practice). Confirm specificity with CEP70 knockout material or immunogen-peptide preabsorption (catalog antibody caption: peptide-preabsorbed negative control); quench endogenous peroxidase and distinguish neuronal lipofuscin pigment from chromogenic signal in brain sections (standard IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected SKU A10748-1 caption does not state a fixative (catalog antibody caption: fixative unreported). Its paraffin-section example used high-pressure, high-temperature Tris-EDTA retrieval at pH 8.0, with antibody at 1:100 overnight at 4°C; retrieval dependence beyond that example is unreported (catalog antibody caption). IF/ICC may make centrosomal or ciliary puncta easier to resolve than chromogenic tissue IHC (UniProt Q8NHQ1: centrosome localization; HPA: supported centrosome and approved primary cilium localization), but neuronal lipofuscin autofluorescence requires attention in brain IF (standard IF practice).
HPA tissue IHC evidence for CEP70
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — 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 |
|---|---|---|---|---|
| Caudate | Neuronal cells | High | Protein (IHC) | HPA → |
| Cerebellum | Purkinje cells | High | Protein (IHC) | HPA → |
| Cerebral cortex | Neuronal cells | High | Protein (IHC) | HPA → |
| Kidney | Cells in tubules | High | Protein (IHC) | HPA → |
| Pancreas | Exocrine glandular cells | High | Protein (IHC) | HPA → |
Section 3
Advanced CEP70 IHC Tips
Troubleshoot CEP70 staining in paraffin sections by checking retrieval, cell type, subcellular pattern and controls before interpreting chromogenic signal.
Which retrieval condition should I start with for CEP70 paraffin-section IHC?
Start with heat-mediated Tris-EDTA at pH 8.0 for CEP70 paraffin sections (datasheet A10748-1). The selected brain-section example used high pressure and temperature for retrieval, then antibody at 1:100 overnight at 4°C (caption A10748-1). If staining is weak, compare a shorter and longer heating cycle on adjacent sections while holding antibody concentration and detection constant (general IHC practice). Check tissue integrity and background alongside signal: excessive heating can damage morphology, making a small centrosomal focus harder to distinguish from chromogen deposits (general IHC practice; UniProt Q8NHQ1: centrosomal localization).
How should I troubleshoot fixation-related loss of CEP70 staining?
The CEP70 brain-section caption identifies paraffin embedding but does not state the fixative, so target-specific fixation sensitivity is unknown (caption A10748-1). Do not infer fixation tolerance from tissue staining patterns or the absence of an annotated transmembrane segment or glycosylation sites (HPA: tissue IHC profile; UniProt Q8NHQ1: topology and glycosylation). Compare sections with documented fixation histories using the same Tris-EDTA pH 8.0 retrieval and detection run (datasheet A10748-1; general IHC practice). Include a known staining section and a no-primary control; interpret differences only after checking morphology, section adhesion and background (general IHC practice).
Where should convincing CEP70 signal appear in tissue sections?
Expect a cytoplasmic component and assess whether discrete perinuclear or spindle-pole-associated foci can be resolved within well-preserved cells (HPA: ubiquitous cytoplasmic tissue expression; UniProt Q8NHQ1: centrosome and mitotic spindle poles). CEP70 interacts directly with tubulin-gamma, which determines its centrosomal localization (UniProt Q8NHQ1: subunit). A chromogenic section may not resolve a single centrosome, so judge focal staining against cytoplasmic pattern, cell morphology and matched controls rather than requiring a visible punctum in every cell (general IHC practice). Diffuse nuclear-only staining warrants caution because HPA calls nucleoplasmic localization uncertain, although its cell-imaging data also support centrosomal localization (HPA: subcellular localization).
Can this IHC stain distinguish CEP70 isoforms or epitope loss?
CEP70 has 5 annotated isoforms, but the supplied IHC caption does not map the antibody epitope to any isoform (UniProt Q8NHQ1: isoforms; caption A10748-1). Therefore, score staining as CEP70 immunoreactivity without assigning an isoform from its intensity or compartment (general IHC interpretation). The caption reports a negative control obtained by preabsorbing antibody with immunogen peptide, which helps assess peptide-dependent staining but does not establish isoform selectivity (caption A10748-1; general IHC practice). If a specimen stains unexpectedly weakly, compare retrieval and preservation with a control section before proposing epitope loss or splice variation (datasheet A10748-1; general IHC practice).
How can IF help evaluate an ambiguous CEP70 IHC pattern?
Use IF on a separately prepared, appropriately controlled specimen to assess whether CEP70 signal overlaps a marker for the expected cell type; HPA reports high tissue staining in cerebellar Purkinje cells and kidney tubular cells (HPA: tissue IHC). Choose fluorophores away from strong tissue autofluorescence and inspect single-channel images before interpreting overlap (general IF practice). CEP70 lacks an annotated transmembrane segment and is associated with cytoplasm and centrosomes, so choose permeabilisation appropriate for an intracellular epitope and verify that it preserves cell structure (UniProt Q8NHQ1: topology and localization; general IF practice). Treat IF localization as supporting evidence alongside the paraffin-section controls, since HPA also reports primary-cilium and basal-body localization (HPA: subcellular localization).
What controls separate CEP70 staining from chromogenic background?
Run a no-primary section to assess detection-reagent background, and include the caption’s peptide-preabsorbed control when reproducing that antibody setup (general IHC practice; caption A10748-1). Block endogenous peroxidase before DAB detection, then compare signal in matching tissue compartments under identical development conditions (general chromogenic IHC practice). The reported brain example used antibody at 1:100 overnight at 4°C; keep those conditions fixed while assessing whether retrieval or detection drives background (caption A10748-1). Diffuse staining across cell-free areas, section folds or tissue edges should be assessed separately from intracellular cytoplasmic and centrosomal patterns expected for CEP70 (general IHC practice; UniProt Q8NHQ1: localization).
How should I score CEP70 IHC across samples? ⚠ ANSWER MARKED FOR VERIFICATION
Predefine the scored cell population and compartment, then report an H-score or the percentage of positive cells for cytoplasmic staining (general IHC quantification practice; HPA: ubiquitous cytoplasmic tissue expression). If discrete foci are reproducibly resolved, report focus-positive cells or focus density per mm² of viable tissue as a separate measure (general IHC quantification practice; UniProt Q8NHQ1: centrosomal localization). Normalize counts to the number of evaluable cells or viable tissue area, and apply one threshold and imaging setup across sections (general IHC quantification practice). Record tissue type and cell identity because HPA reports high staining in kidney tubular cells but no detection in adipocytes (HPA: tissue IHC).
When is a CEP70-positive IHC result convincing?
A convincing result has reproducible intracellular staining in morphologically intact cells, with a cytoplasmic pattern and, where resolvable, centrosomal foci (HPA: tissue IHC profile; UniProt Q8NHQ1: localization). Check cell identity against tissue context: high staining is reported in cerebral-cortex neuronal cells, whereas adipocytes are reported as not detected (HPA: tissue IHC). Treat isolated edge staining, necrotic areas, extracellular deposits and signal remaining in a no-primary section as possible artefacts or detection background (general IHC practice). Strong nuclear-only staining needs independent support because HPA labels nucleoplasmic localization uncertain; compare the peptide-preabsorbed control before attributing such signal to CEP70 (HPA: subcellular localization; caption A10748-1).
Boster reagents
Best CEP70 / Centrosomal protein of 70 kDa IHC Antibodies
The catalog lists anti-CEP70 antibody A10748-1 for IHC and IF in human and monkey (catalog: applications and reactivity); its figure documents IHC on a human brain paraffin section (IHC caption).
Real IHC data
Anti-Centrosomal protein of 70 kDa CEP70 Antibody
Cat # A10748-1
A10748-1 will render with an IHC figure from a human brain paraffin section, stained at 1:100 after Tris-EDTA pH 8.0 retrieval (IHC caption). IF is listed as an application, and human and monkey are listed as reactive species; no IF figure is supplied (catalog: applications, reactivity, and image captions).
Which to pick: For paraffin-section tissue IHC, choose A10748-1: its own caption documents human brain staining, but does not report the fixative (IHC caption). For IF, A10748-1 is listed at 1:50; ICC is not listed, no IF figure is supplied, and clonality is unreported (catalog: applications, dilution, image captions, and clone field). A10748-1 also lists monkey reactivity, though its IHC figure documents human tissue only (catalog: reactivity; IHC caption).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q8NHQ1 (CEP70_HUMAN, Centrosomal protein of 70 kDa).
- Human Protein Atlas. CEP70 tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. CEP70 subcellular location (ICC-IF): Mainly localized to the nucleoplasm, cytosol and primary cilium. In addition localized to the centrosome and basal body..
- Human Protein Atlas. CEP70 antibody validation summary (2 antibodies).
- Cep70 overexpression stimulates pancreatic cancer by inducing centrosome abnormality and microtubule disorganization. Scientific reports 2016 — PMC4759539.
- PubMed PMID:14702039 — UniProt-cited evidence.
- PubMed PMID:16641997 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for CEP70 (Centrosomal protein of 70 kDa). Expected localisation Cytoplasmic in tissue sections (HPA tissue IHC). Antigen retrieval: Tris-EDTA pH 8.0 HIER, heat-mediated (datasheet A10748-1). Validated IHC antibody: A10748-1 at 1:100-1:300 (the product's own IHC validation image is shown). Controls (HPA): positive Caudate (Neuronal cells); negative Adipose tissue. IHC workflow; specimen preparation unspecified. Protocol steps: 1. Sample: Paraffin-embedded Human brain tissue; fixative not specified (datasheet A10748-1); 2. Fixation: Image fixative and duration unreported (datasheet A10748-1); 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: Tris-EDTA pH 8.0 (datasheet A10748-1); 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-CEP70, 1:100-1:300 (datasheet A10748-1); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep paraffin tissue fixation consistent across samples. (standard IHC practice; not target-specific); Caveat: Adipocytes and cholangiocytes may lack staining (HPA tissue IHC); Isoform / epitope: 5 isoforms; epitope coverage is unspecified (UniProt); Regulation: Low tissue specificity (HPA tissue RNA). Evidence: UniProt Q8NHQ1, HPA tissue IHC · Enhanced, HPA ICC-IF · Mainly localized to the nucleoplasm, cytosol and primary cilium. In…, Datasheet A10748-1, PMC4759539. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/CEP70/immunohistochemistry-ihc-guide.