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CEP70 / Centrosomal protein of 70 kDa · IHC design guide
Design Immunohistochemistry for CEP70
Plan CEP70 paraffin-section IHC around its cytoplasmic tissue staining (HPA tissue IHC). The catalog antibody has an IHC dilution range of 1:100–1:300 (datasheet A10748-1); compare sections with consistent fixation and appropriate negative controls (standard IHC practice).
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 tissue staining (HPA tissue IHC); centrosomal localization (UniProt) | |
| Staining pattern | Cytoplasmic staining across diverse tissue 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 fixation consistent between sections (standard IHC practice; not target-specific) | |
| Caveat | Staining and RNA expression show medium consistency (HPA tissue IHC) | |
| Regulation | No expression regulation annotated (UniProt) | |
| Isoform / epitope | 5 isoforms; verify epitope coverage across variants (UniProt) |
Section 1
Recommended CEP70 IHC & IF Protocols
The catalog antibody protocol (datasheet: A10748-1) is accompanied by one published CEP70 IHC protocol for pancreatic tissues (PMC4759539: Immunohistochemistry).
| 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 retrieval at pH 8.0 (datasheet: A10748-1); the published protocol confirms retrieval but does not specify its conditions (PMC4759539: Immunohistochemistry).
Section 2
What Is the Expected CEP70 Staining Pattern?
CEP70 is a centrosome-associated cytoplasmic protein with no transmembrane segment (UniProt Q8NHQ1: location and topology). In paraffin-section IHC, expect predominantly cytoplasmic staining across many cell types (HPA tissue IHC: ubiquitous cytoplasmic expression; reliability Enhanced). High staining is reported in neuronal cells, Purkinje cells, kidney tubules and several other specified cell populations (HPA tissue IHC: High).
| Cytoplasmic signal in kidney tubular cells, with weaker signal in some surrounding cells. | This fits a reported high-staining population within a broadly cytoplasmic tissue profile (HPA tissue IHC: kidney tubular cells High; ubiquitous cytoplasmic expression). Judge intensity by cell type and local background; a centrosomal focus may be too small to resolve confidently in chromogenic sections (UniProt Q8NHQ1: centrosome localisation; general IHC practice). |
| Strong, exclusively nuclear chromogen with little cytoplasmic signal. | Treat this as discordant with the tissue IHC profile and investigate staining artefact (HPA tissue IHC: ubiquitous cytoplasmic expression; general IHC practice). HPA also reports uncertain nucleoplasmic localisation by ICC-IF, so nuclear signal alone cannot settle whether an IF observation is specific (HPA subcellular: nucleoplasm uncertain). |
| Prominent staining in adipocytes or liver cholangiocytes. | These particular cell populations were reported as not detected (HPA tissue IHC: adipocytes and cholangiocytes Not detected). Recheck cell identification, background and detection controls before assigning CEP70 positivity; cross-reactivity or endogenous detection activity is possible, but the HPA examples do not make either tissue universally negative (general IHC practice; HPA tissue IHC: specified cells Not detected). |
| Diffuse chromogen obscures cell borders or appears similarly in tissue and control sections. | A uniform haze cannot establish the expected cellular distribution (HPA tissue IHC: cytoplasmic profile; general IHC practice). Inspect a section processed without primary antibody and assess blocking, washing and detection chemistry; the pattern may reflect nonspecific binding or endogenous activity (general IHC practice). |
| No convincing signal in kidney tubular cells or cerebellar Purkinje cells. | Both are reported high-staining populations, so absent signal warrants a run-level check (HPA tissue IHC: kidney tubular cells and Purkinje cells High). Review tissue identity, the antibody's IHC validation, detection controls and the recorded retrieval procedure before interpreting a specimen as negative (HPA antibodies: IHC Enhanced; general IHC practice). |
💡Expected CEP70 appearanceCall a slide positive when identifiable cells show predominantly cytoplasmic staining, including strong signal in an HPA high-staining population; uniform haze or isolated strong staining in reported not-detected cells calls for control review (HPA tissue IHC: cytoplasmic profile, High and Not detected populations; general IHC practice).
| Cell population and tissue | The profile is broadly cytoplasmic, yet staining levels differ by cell population (HPA tissue IHC: ubiquitous cytoplasmic expression; specified High, Low and Not detected cells). Compare equivalent cells rather than treating every part of a section as an equal-intensity reference (general IHC practice). |
| Subcellular scale | CEP70 is reported at the centrosome and mitotic spindle poles (UniProt Q8NHQ1: subcellular location). A discrete centrosomal dot may be difficult to distinguish from chromogenic background, so cytoplasmic distribution and cell identity remain useful IHC checks (general IHC practice). |
| Antibody evidence and isoforms | Two listed rabbit polyclonal antibodies have Enhanced IHC status, while their ICC status is Uncertain (HPA antibodies: HPA036941 and HPA036942). Five isoforms are listed, but the payload gives no epitope or isoform-coverage data; do not infer that either antibody detects every isoform (UniProt Q8NHQ1: five isoforms; HPA antibodies). |
| IF/ICC Q: what localisation should be expected? | A: HPA reports primary cilium and basal body localisation as approved, centrosome as supported, and nucleoplasm and cytosol as uncertain (HPA subcellular: ICC-IF). Interpret IF/ICC on its own evidence; those location labels do not establish an IHC-P staining pattern (HPA subcellular: ICC-IF; HPA tissue IHC: cytoplasmic profile). |
| Retrieval and processing evidence | No target-specific fixation or antigen-retrieval effect is supplied; tissue intensity and the lack of a transmembrane segment cannot predict either effect (HPA tissue IHC: staining levels; UniProt Q8NHQ1: topology). Record the actual retrieval conditions when comparing IHC runs (general IHC practice). |
| Situation | Likely cause | Next action |
|---|---|---|
| Expected high-staining cells are blank. | A failed staining run, incorrect tissue identification or unsuitable antibody conditions may explain absent signal (HPA tissue IHC: specified High populations; general IHC practice). | Verify the cell population and include a known positive section; review the IHC-validated antibody's documented conditions, retrieval record and detection controls before calling the test section negative (HPA antibodies: IHC Enhanced; general IHC practice). |
| Every compartment shows similar brown haze. | Nonspecific antibody binding or detection background can conceal the reported cytoplasmic pattern (HPA tissue IHC: cytoplasmic expression; general IHC practice). | Compare with a no-primary control; check blocking, wash steps and detection chemistry, then reassess cellular localisation in the cleared section (general IHC practice). |
| Adipocytes or cholangiocytes stain more strongly than nearby expected cells. | Cell misidentification, cross-reactivity or endogenous detection activity is possible (HPA tissue IHC: adipocytes and cholangiocytes Not detected; general IHC practice). | Confirm morphology and compare a no-primary control with an HPA high-staining cell population. Treat these reported negatives as cell-specific reference observations, not absolute negative-tissue controls (HPA tissue IHC: specified Not detected and High populations; general IHC practice). |
| Signal appears only nuclear. | This differs from the reported tissue IHC profile; uncertain nucleoplasmic ICC-IF localisation does not validate nuclear-only chromogenic staining (HPA tissue IHC: cytoplasmic expression; HPA subcellular: nucleoplasm uncertain). | Inspect background controls and compare the same run with a high-staining tissue population. Report the discordance if it persists rather than scoring it as the expected IHC result (HPA tissue IHC: specified High populations; general IHC practice). |
| Signal varies markedly between sections or runs. | Section quality, retrieval execution or detection variation can change an IHC readout (general IHC practice); no CEP70-specific fixation sensitivity is established by the supplied sources. | Compare matched cell populations and a common positive control across runs, then review the recorded processing and detection steps (HPA tissue IHC: cell-specific levels; general IHC practice). |
| ICC-IF puncta seem inconsistent with diffuse IHC staining. | The assays resolve different spatial detail, while HPA assigns different confidence levels to its ICC-IF locations (HPA subcellular: location labels; general IHC practice). | Assess each assay against its own reported pattern: cytoplasmic tissue IHC and the qualified ciliary, centrosomal, basal-body, cytosolic and nucleoplasmic ICC-IF observations (HPA tissue IHC: cytoplasmic expression; HPA subcellular: ICC-IF). |
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 cerebral cortex first; neuronal cells should stain (HPA: High in cerebral cortex neuronal cells). Run adipose tissue as the negative tissue because adipocytes are not detected (HPA: Not detected in adipocytes); the cerebral cortex row establishes no internal negative cell type, so neighboring unstained cells are a background reference, not a verified biological negative (HPA: cerebral cortex neuronal-cell row).
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 matched to the primary antibody’s host species and immunoglobulin class, with monoclonal or polyclonal format matched where feasible (standard IHC practice). Use CEP70 knockout material or immunogen-peptide preabsorption as a biological specificity control (A10748-1 caption: peptide-preabsorbed negative control), and block endogenous peroxidase before chromogenic detection (standard IHC practice).
⚠️Feasibility: No target-specific fixation window or fixation effect is reported, and the selected A10748-1 human-brain paraffin caption does not state a fixative (A10748-1 caption: fixative unreported). The caption reports high-pressure, high-temperature Tris-EDTA retrieval at pH 8.0 and antibody incubation at 1:100 overnight at 4°C; whether retrieval is required has not been established by a comparison (A10748-1 caption). The supplied evidence does not establish that frozen sections or IF are easier; brain lipofuscin may cause IF autofluorescence (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, controls, cellular distribution and scoring before interpreting weak or diffuse signal.
Which retrieval condition should I start with for CEP70 paraffin IHC?
Start with heat-mediated Tris-EDTA retrieval at pH 8.0 for paraffin sections (datasheet A10748-1). The selected human-brain image used high-pressure, high-temperature retrieval in that buffer, followed by primary antibody at 1:100 overnight at 4°C (caption A10748-1). If staining is weak, compare a shorter and longer heating cycle on matched sections while keeping detection and imaging conditions constant (standard IHC practice). Check tissue integrity and background alongside signal; excessive heating can damage morphology and make apparent cellular staining harder to interpret (standard IHC practice).
How should I assess whether fixation is suppressing CEP70 staining?
The selected caption identifies paraffin-embedded human brain but does not state its fixative, so CEP70 sensitivity to a particular fixation condition remains unknown (caption A10748-1). Record fixative, fixation duration and processing history for each specimen, then compare matched sections using the same pH 8.0 Tris-EDTA retrieval (datasheet A10748-1; standard IHC practice). Include a concurrently processed positive tissue and a no-primary control to distinguish lost antigen signal from detection background (standard IHC practice). Do not assign weak staining to fixation alone when section quality, retrieval, antibody exposure and tissue preservation also vary (standard IHC practice).
What staining pattern should support a CEP70 call in tissue?
Assess cytoplasmic signal in its cellular context: tissue IHC is described as ubiquitously cytoplasmic, although staining and RNA show medium consistency (HPA tissue IHC). Centrosomal localization is biologically plausible because CEP70 binds tubulin-gamma and occurs at the center of interphase microtubule arrays and at mitotic spindle poles (UniProt Q8NHQ1). A small centrosomal focus may be difficult to resolve in chromogenic sections, so avoid requiring a visible punctum in every positive cell (UniProt Q8NHQ1; standard IHC practice). Review nuclear or ciliary staining cautiously: those locations come from cell-based imaging and have different confidence assignments (HPA subcellular).
Could isoform differences explain inconsistent CEP70 staining?
CEP70 has 5 annotated isoforms, but the supplied record does not map the catalog antibody’s epitope to any of them (UniProt Q8NHQ1; supplied product evidence). Ask whether the immunogen sequence is shared across isoforms before treating staining differences as altered CEP70 abundance (UniProt Q8NHQ1; standard IHC practice). The selected image includes a peptide-preabsorbed negative control, which supports testing antigen-dependent staining for that specimen but does not establish isoform specificity (caption A10748-1). Compare matched sections and, where available, an independently characterized antibody against a distinct epitope before attributing a compartment shift to alternative splicing (standard IHC practice).
How can IF help resolve an ambiguous CEP70 IHC pattern?
Use IF as a separate localization check, pairing CEP70 with a marker for the cell type under examination and, if needed, a centrosome marker (standard IF practice; UniProt Q8NHQ1). Select spectrally separated fluorophores after checking unstained tissue for autofluorescence, and acquire single-stain controls to assess bleed-through (standard IF practice). CEP70 lacks a transmembrane segment and is associated with cytoplasmic and centrosomal structures, so test gentle permeabilisation for access to intracellular epitopes without assuming the antibody’s epitope is known (UniProt Q8NHQ1; standard IF practice). Compare puncta with the reported centrosome, basal-body and primary-cilium locations, while keeping IF findings distinct from chromogenic IHC scoring (HPA subcellular; standard IF practice).
How do I reduce diffuse or granular background in CEP70 IHC?
First compare the stained section with a no-primary control and the peptide-preabsorbed control shown for the selected human-brain image (caption A10748-1; standard IHC practice). If background persists, optimize blocking and washing, then titrate the primary around the documented 1:100 overnight condition while holding retrieval constant (caption A10748-1; standard IHC practice). For peroxidase detection, include a peroxidase block and inspect pigment or tissue deposits before assigning granular DAB signal to CEP70 (standard IHC practice). Retain cellular context: broad cytoplasmic staining can be compatible with the tissue profile, so diffuse signal alone does not prove nonspecific binding (HPA tissue IHC).
What is a defensible way to quantify CEP70 chromogenic staining? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and anatomical region before scoring, then report percent positive cells and an H-score combining intensity with the proportion at each intensity (standard IHC practice). Normalize counts to eligible cells or area, such as positive cells per mm², and use the same thresholds and exposure-independent viewing conditions across sections (standard IHC practice). Score cytoplasmic staining separately from discrete puncta or nuclear signal because these patterns have different localization support (HPA tissue IHC; HPA subcellular; UniProt Q8NHQ1). Stratify by cell type when comparing samples: high staining is reported in neuronal cells and kidney tubule cells, whereas adipocytes were not detected (HPA tissue IHC).
When should an apparent CEP70-positive cell be considered an artefact?
Treat a sharply confined section-edge signal, necrotic debris or staining reproduced in a no-primary control as suspect before calling a cell positive (standard IHC practice). Check for residual endogenous peroxidase activity when using DAB, and compare the suspect area with intact neighboring tissue under identical detection conditions (standard IHC practice). Cytoplasmic staining fits the tissue profile, while centrosomal localization fits the protein record; isolated nuclear staining needs additional corroboration because that cell-based assignment is uncertain (HPA tissue IHC; UniProt Q8NHQ1; HPA subcellular). Consider cell identity: neuronal cells are reported high in several brain regions, but morphology and control behavior still govern each call (HPA tissue IHC; standard IHC practice).
Boster reagents
Best CEP70 / Centrosomal protein of 70 kDa IHC Antibodies
A10748-1 has a human brain paraffin-section IHC image (IHC image caption); IF is listed without an image (catalog: applications and image data). Human and Monkey reactivity is listed (catalog: reactivity).
Real IHC data
Anti-Centrosomal protein of 70 kDa CEP70 Antibody
Cat # A10748-1
A10748-1 has an IHC image from paraffin-embedded human brain, with a peptide-preabsorbed control (IHC image caption). The same SKU lists IF as an application and Human and Monkey reactivity, but provides no IF image (catalog: applications, reactivity, and image data).
Which to pick: Choose A10748-1 for tissue IHC when using paraffin sections: its human brain image documents that preparation and 1:100 antibody dilution with Tris-EDTA pH 8.0 retrieval; the fixative is unreported (IHC image caption). For IF, A10748-1 lists 1:50, but IF/ICC sample validation is unreported (catalog: IF dilution and image data). For cross-species work, A10748-1 lists Human and Monkey reactivity, while its IHC image documents only human tissue; the host is rabbit and clonality is unreported (catalog: reactivity, host, and clone; IHC image 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 tissue staining (HPA tissue IHC); centrosomal localization (UniProt). 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 fixation consistent between sections (standard IHC practice; not target-specific); Caveat: Staining and RNA expression show medium consistency (HPA tissue IHC); Isoform / epitope: 5 isoforms; verify epitope coverage across variants (UniProt); Regulation: No expression regulation annotated (UniProt). 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.