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CIAPIN1 / Anamorsin · IHC design guide
Design Immunohistochemistry for CIAPIN1
Plan CIAPIN1 paraffin-section IHC around predominantly cytoplasmic staining, with nuclear staining in some tissues (HPA tissue IHC). Use testis pachytene spermatocytes as a strong reference and assess antibody staining alongside controls (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
CIAPIN1 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; nuclear in some tissues (HPA tissue IHC) | |
| Staining pattern | Mostly cytoplasmic, with nuclear staining in some tissues (HPA tissue IHC) | |
| Antigen retrieval | Tris-EDTA pH 9.0 HIER, 95–98 °C, 20 min (rule: nuclear antigen) | |
| Positive control | Testis+4 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 paraffin sections. (standard IHC practice; not target-specific) | |
| Caveat | Antibody staining and RNA show medium consistency (HPA tissue IHC) | |
| Regulation | Tissue abundance varies; testis is highest (HPA tissue IHC) | |
| Isoform / epitope | 3 isoforms; epitope coverage is unspecified (UniProt) |
Section 1
Recommended CIAPIN1 IHC & IF Protocols
Compare the catalog antibody’s IHC-P protocol (datasheet) with four published CIAPIN1 tissue IHC protocols (PMC6481027; PMC9267148; PMC3502349; PMC2516489).
| Sample | Paraffin-embedded human liver injury tissue; fixative not specified (datasheet A04333) |
| Fixation | Image fixative and duration unreported (datasheet A04333); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat-induced epitope retrieval in Tris-EDTA buffer, pH 9.0, 20 min at 95–98 °C (standard rule: nuclear antigen) |
| 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-CIAPIN1, 1:50-1:200 (datasheet A04333) |
| 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 | CIAPIN1-positive staining in pachytene spermatocytes of testis (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression in most tissues which in some tissues was combined with nuclear expression, most abundant in testis. No signal in the no-primary control. |
💡Decision noteStart with Tris-EDTA pH 9.0 HIER at 95–98 °C for 20 min (page retrieval rule: nuclear antigen). Citrate pH 6.0 is a published alternative (PMC3502349).
Section 2
What Is the Expected CIAPIN1 Staining Pattern?
CIAPIN1 is mainly cytoplasmic by tissue IHC, with nuclear staining in some tissues; UniProt also places it in the mitochondrial intermembrane space and reports no transmembrane segment (HPA tissue IHC; UniProt Q6FI81 localization and topology). Expect strongest staining in testis pachytene spermatocytes and moderate staining in several glandular cell populations (HPA tissue IHC). HPA rates the tissue IHC evidence Enhanced, with medium consistency between staining and RNA expression (HPA tissue IHC reliability).
| Pachytene spermatocytes show strong cytoplasmic staining; some nuclei also stain. | This fits the strongest listed tissue result and the reported cytoplasmic pattern with nuclear staining in some tissues (HPA tissue IHC). Judge nuclear staining alongside the cytoplasmic signal and tissue context; nuclear signal alone does not reproduce the typical tissue pattern (HPA tissue IHC). |
| Staining is confined to cell borders or extracellular spaces, with little intracellular signal. | This is inconsistent with the reported cytoplasmic and occasional nuclear tissue pattern and with the absence of a transmembrane segment (HPA tissue IHC; UniProt Q6FI81 topology). Suspect nonspecific staining or a detection artefact; check a known-positive section and detection controls (general IHC practice). |
| Adipocytes stain strongly while expected positive cells show little signal. | HPA reports adipocytes as not detected in adipose tissue and breast, while testis pachytene spermatocytes are High (HPA tissue IHC). Consider cross-reactivity or endogenous detection activity; interpret the named cell populations, rather than treating every cell in a tissue as positive or negative (general IHC practice; HPA tissue IHC). |
| Most of the section has an even brown haze that obscures cell boundaries. | Uniform haze cannot establish the cell-specific cytoplasmic pattern reported by HPA (HPA tissue IHC). Assess the no-primary control and review blocking, washes, antibody concentration and chromogen development as general IHC troubleshooting steps (general IHC practice); the supplied sources do not identify a CIAPIN1-specific cause. |
| A testis section has no detectable signal in pachytene spermatocytes. | That conflicts with HPA's High result for this cell population (HPA tissue IHC). First confirm the cells are present and the run's detection controls worked; then review the catalog antibody's IHC-P instructions and the run's retrieval and dilution settings (general IHC practice). No target-specific fixation sensitivity is established here. |
💡Expected CIAPIN1 appearanceA convincing positive is High intracellular, mainly cytoplasmic staining in testis pachytene spermatocytes, with possible nuclear staining; uniform haze or cell-border-only signal is suspect (HPA tissue IHC; UniProt Q6FI81 topology).
| Which tissue and cells should anchor interpretation? | Use testis pachytene spermatocytes as a strong reference; HPA calls them High, while appendix and duodenum glandular cells are Medium (HPA tissue IHC). Pancreatic exocrine glandular cells are Low despite UniProt's high pancreas expression statement; distinguish a named cell's IHC level from whole-tissue expression (HPA tissue IHC; UniProt Q6FI81 tissue specificity). |
| How much confidence does the tissue pattern support? | HPA assigns Enhanced IHC validation to HPA041350 and HPA042182, while describing overall staining-to-RNA consistency as medium (HPA antibodies; HPA tissue IHC reliability). Treat this as support for the reported pattern, with controls and cell-level interpretation still needed in a new experiment (general IHC practice). |
| Does topology predict a membrane stain? | UniProt reports no transmembrane segment and lists cytoplasm, nucleus and mitochondrial intermembrane space (UniProt Q6FI81 localization and topology). The tissue IHC profile is predominantly cytoplasmic, sometimes nuclear (HPA tissue IHC). A crisp plasma-membrane outline therefore needs independent validation before assignment to CIAPIN1 (HPA tissue IHC; UniProt Q6FI81 topology). |
| Can the supplied record resolve isoforms or fixation effects? | Target-specific fixation effects are not established by the supplied assay evidence. Verify with a matched IHC source before attributing a result to fixation. |
| Q: What should IF/ICC show? | A: HPA reports mainly mitochondrial localization with additional nucleoplasmic localization, both supported (HPA subcellular ICC-IF). UniProt also lists the mitochondrial intermembrane space, cytoplasm and nucleus (UniProt Q6FI81 localization). This is an interpretation reference for IF/ICC; it does not turn the tissue IHC pattern into a resolved mitochondrial IHC readout. |
| Situation | Likely cause | Next action |
|---|---|---|
| Testis pachytene spermatocytes are weak or blank. | Possible run failure or a mismatch between the applied conditions and the catalog antibody's IHC-P instructions; HPA reports these cells as High (HPA tissue IHC; general IHC practice). | Verify the cell population and run controls, then review the antibody's IHC-P retrieval, dilution and detection instructions (general IHC practice). Do not infer a CIAPIN1-specific fixation problem from this result. |
| A weak signal appears in pancreatic exocrine glandular cells. | That cell population is Low by HPA tissue IHC, although UniProt describes pancreas as highly expressed at the tissue level (HPA tissue IHC; UniProt Q6FI81 tissue specificity). | Compare with a stronger reference such as testis pachytene spermatocytes before calling the run insensitive (HPA tissue IHC; general IHC practice). Record the precise cells scored. |
| Adipocytes show prominent brown signal. | HPA lists adipocytes in adipose tissue and breast as not detected; nonspecific binding or endogenous detection activity are possible general IHC causes (HPA tissue IHC; general IHC practice). | Check a no-primary control and the detection system's endogenous-activity blocking steps; compare the signal's distribution with a positive cell population (general IHC practice; HPA tissue IHC). |
| Staining is mostly nuclear with no discernible cytoplasm. | Nuclear expression can accompany cytoplasmic expression in some tissues, but HPA describes the overall tissue pattern as cytoplasmic (HPA tissue IHC). | Review the matched tissue reference and controls before scoring it as expected; document both compartments separately (HPA tissue IHC; general IHC practice). |
| Staining forms sharp cell-border outlines. | This differs from the reported intracellular tissue pattern and the lack of a transmembrane segment (HPA tissue IHC; UniProt Q6FI81 topology). | Reassess specificity with a known-positive section and detection controls; avoid assigning border signal to CIAPIN1 without further evidence (general IHC practice). |
| Brown background obscures medium-level glandular staining. | Diffuse background can conceal the Medium glandular-cell signals listed for appendix, duodenum, gallbladder, rectum, small intestine and stomach (HPA tissue IHC; general IHC practice). | Check the no-primary control, then review blocking, washes, antibody concentration and chromogen development under the IHC-P workflow (general IHC practice). Score only interpretable cells. |
Experiment evidence
IHC & IF images showing CIAPIN1 staining
A04333 · IHCImmunohistochemistry of paraffin-embedded human liver injury using CIAPIN1 Antibody.
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000005194-CIAPIN1/tissue/cerebral+cortex#imid_11020575
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000005194-CIAPIN1/tissue/colon#imid_11020509
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000005194-CIAPIN1/tissue/liver#imid_11020494
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000005194-CIAPIN1/tissue/kidney#imid_11020495
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA041350; green = CIAPIN1, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA041350; green = CIAPIN1, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for CIAPIN1 IHC & IF
🧪Run testis first: pachytene spermatocytes should stain (HPA: High in pachytene spermatocytes). Use adipose tissue as the negative comparator because adipocytes are not detected (HPA: Not detected in adipocytes); on the testis slide, assess staining in other identifiable cells as an internal background reference without assuming they are CIAPIN1-negative (HPA: High in pachytene spermatocytes).
Positive control tissue: Testis (Pachytene spermatocytes, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show CIAPIN1 in A-431, U-251MG, U2OS, with annotated localisation: Mitochondria (supported) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control and an isotype control matched to the primary antibody’s host species and clonality; use CIAPIN1 knockout material or a validated peptide-block condition as a biological specificity control (standard IHC practice). For chromogenic testis IHC, quench endogenous peroxidase and inspect unstained or control sections for background; for IF, assess tissue autofluorescence (standard IHC/IF practice).
⚠️Feasibility: A CIAPIN1-specific fixation window and retrieval dependency are unreported in the supplied evidence; optimize antigen retrieval empirically (supplied evidence; standard IHC practice). The selected A04333 caption documents paraffin-section IHC in human liver injury but does not report the fixative or establish fixation (selected tissue-IHC caption). Frozen sections are not shown to be easier here; ICC-IF images exist for A-431, U-251MG, and U2OS, while dense testis cell populations require careful morphology and counterstain review when scoring pachytene spermatocytes (HPA subcellular; HPA: High in pachytene spermatocytes; standard IHC practice). The selected A04333 tissue-IHC caption documents paraffin sections, but does not specify the fixative (selected-SKU IHC image A04333).
HPA tissue IHC evidence for CIAPIN1
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 |
|---|---|---|---|---|
| Testis | Pachytene spermatocytes | High | Protein (IHC) | HPA → |
| Appendix | Glandular cells | Medium | Protein (IHC) | HPA → |
| Duodenum | Glandular cells | Medium | Protein (IHC) | HPA → |
| Gallbladder | Glandular cells | Medium | Protein (IHC) | HPA → |
| Ovary | Follicle cells | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Breast | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Caudate | Glial cells | Not detected | Protein (IHC) | HPA → |
| Cerebellum | Cells in granular layer | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced CIAPIN1 IHC Tips
Troubleshoot CIAPIN1 staining in paraffin sections by checking retrieval, compartment pattern, controls and scoring before interpreting biological differences.
What retrieval should I try when CIAPIN1 staining is weak in paraffin sections?
Start with Tris-EDTA at pH 9.0, heated to 95–98 °C for 20 min (page retrieval setting). Run matched control sections alongside the test sections, then compare staining intensity and tissue preservation under the same antibody and detection conditions (standard IHC practice). If staining remains weak, test a shorter or longer heating interval on adjacent sections before considering another buffer, because retrieval can expose epitopes while damaging morphology (standard IHC practice). Record cytoplasmic and nuclear staining separately: tissue IHC shows mostly cytoplasmic expression, sometimes with nuclear staining (HPA tissue IHC profile).
How can I assess whether fixation caused weak CIAPIN1 staining?
CIAPIN1-specific fixation sensitivity is unknown from the supplied evidence; the selected paraffin-section caption does not state a fixative (A04333 tissue-IHC caption). Record the fixative, fixation duration and processing history for each specimen, and compare adjacent sections processed together when investigating a weak result (standard IHC practice). Use the same retrieval, antibody incubation and chromogenic detection across that comparison so processing differences are easier to interpret (standard IHC practice). Treat a processing-associated signal change as an assay observation until it is reproduced; neither the reported tissue pattern nor the protein’s topology establishes a fixation effect (HPA tissue IHC profile; UniProt Q6FI81 topology).
Should CIAPIN1 appear in nuclei, cytoplasm or mitochondria on chromogenic IHC?
Score cytoplasmic and nuclear staining separately: tissue IHC reports cytoplasmic expression in most tissues, sometimes combined with nuclear expression (HPA tissue IHC profile). UniProt also lists cytoplasm, nucleus and mitochondrial intermembrane space, while cell imaging mainly localizes CIAPIN1 to mitochondria with additional nucleoplasmic signal (UniProt Q6FI81 subcellular record; HPA subcellular). At routine brightfield resolution, a granular cytoplasmic pattern alone cannot establish mitochondrial localization, so avoid assigning an organelle from DAB staining alone (standard IHC interpretation). Check whether the compartment pattern repeats in intact cells and matched sections before treating an isolated nuclear or diffuse signal as biological variation (standard IHC practice).
Can differences between sections reflect CIAPIN1 isoforms or epitope masking?
Three CIAPIN1 isoforms are listed, but the supplied evidence does not map this antibody’s epitope to any isoform (UniProt Q6FI81 isoforms; A04333 tissue-IHC caption). Do not attribute a staining difference to isoform expression unless an independently validated epitope map or isoform-specific control supports that claim (standard IHC interpretation). The record lists phosphorylation sites including residues 136, 177 and 182, but does not establish that these modifications affect antibody binding (UniProt Q6FI81 modified residues). Compare retrieval conditions and antibody controls on adjacent sections first, and report the observed compartment and cell type rather than assigning an untested molecular cause (standard IHC practice).
How should I check a chromogenic CIAPIN1 pattern with multiplex IF?
Use IF as a separate validation experiment and pair CIAPIN1 with a marker for the cell population being scored, such as pachytene spermatocytes when examining testis (HPA tissue IHC: high in pachytene spermatocytes). Choose fluorophores after checking unstained tissue for autofluorescence, and include single-stain controls to assess channel bleed-through (standard IF practice). CIAPIN1 has no transmembrane segment and is reported in intracellular compartments, so permeabilize sufficiently for access to cytoplasmic, nuclear or mitochondrial epitopes; the antibody’s exact epitope location is unspecified (UniProt Q6FI81 topology and subcellular record; standard IF practice). Compare cell identity and compartment pattern with chromogenic sections, while treating mitochondrial colocalization as an IF observation rather than proof from DAB alone (HPA subcellular; standard IHC/IF interpretation).
How do I separate CIAPIN1 staining from nonspecific brown background?
Include a section without primary antibody and inspect it for endogenous peroxidase activity or nonspecific detection-system staining (standard chromogenic IHC practice). Apply a peroxidase block before DAB development and keep development time consistent across test and control sections (standard chromogenic IHC practice). Evaluate diffuse pigment, precipitate and staining at damaged section edges apart from signal confined to intact cells; CIAPIN1 tissue IHC is predominantly cytoplasmic, with nuclear expression in some tissues (HPA tissue IHC profile; standard IHC interpretation). If background persists, adjust antibody concentration or blocking conditions using matched sections and retain the condition that preserves the expected cellular pattern (standard IHC practice).
How should I quantify CIAPIN1 across regions with different cellular composition? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring, then report the percentage of positive cells and an intensity-weighted H-score when staining intensity is reproducible (standard IHC quantification). An H-score uses intensity categories from 0–3 and ranges from 0–300 when each category’s percentage is multiplied by its score (standard IHC quantification). Normalize counts to the number of eligible intact cells, or report positive-cell density per mm² of viable tissue when area is the relevant denominator (standard IHC quantification). Keep cytoplasmic and nuclear scores separate, and compare like cell populations because the reported tissue pattern varies by cell type (HPA tissue IHC profile).
What makes a positive CIAPIN1 IHC result convincing?
A convincing result stains intact cells in a reproducible compartment pattern, with cytoplasmic signal and possible nuclear signal consistent with tissue IHC observations (HPA tissue IHC profile; standard IHC interpretation). Check cell identity against the section: pachytene spermatocytes show high staining in testis, whereas adipocytes are listed as not detected in adipose tissue (HPA tissue IHC). Treat staining limited to cut edges, necrotic areas or the no-primary control as suspect, and check endogenous peroxidase before assigning DAB signal to CIAPIN1 (standard chromogenic IHC practice). Because HPA reports only medium consistency between staining and RNA data, interpret a new pattern alongside controls and tissue morphology (HPA reliability description; standard IHC practice).
Boster reagents
Best CIAPIN1 / Anamorsin IHC Antibodies
A04333 lists IHC and IF use in human and mouse samples (catalog: applications/reactivity). Its supplied image shows IHC in paraffin-embedded human liver-injury tissue (A04333 image alt); no IF image is supplied (catalog: IF image alts).
Real IHC data
Anti-Anamorsin CIAPIN1 Antibody
Cat # A04333
A04333 is the only card (catalog: SKU), with an IHC image of paraffin-embedded human liver-injury tissue (A04333 image alt). IF is listed and human/mouse reactivity is claimed (catalog: applications/reactivity), but no IF or mouse image is supplied (catalog: image alts).
Which to pick: For tissue IHC, choose A04333: its own caption documents paraffin-embedded human liver-injury tissue, but does not report the fixative (A04333 image alt). For IF, A04333 lists a 1:50–1:200 dilution (catalog: IF dilution); ICC validation and an IF image are unreported (catalog: applications/IF image alts). A04333 lists human and mouse reactivity, has a rabbit host, and does not specify clonality (catalog: reactivity/host/clone); the supplied IHC image documents human tissue only (A04333 image alt).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q6FI81 (CPIN1_HUMAN, Anamorsin).
- Human Protein Atlas. CIAPIN1 tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. CIAPIN1 subcellular location (ICC-IF): Mainly localized to the mitochondria. In addition localized to the nucleoplasm..
- Human Protein Atlas. CIAPIN1 antibody validation summary (2 antibodies).
- CIAPIN1 Targeted NHE1 and ERK1/2 to Suppress NSCLC Cells' Metastasis and Predicted Good Prognosis in NSCLC Patients Receiving Pulmonectomy. Oxidative medicine and cellular longevity 2019 — PMC6481027.
- Prediction of CIAPIN1 (Cytokine-Induced Apoptosis Inhibitor 1) Signaling Pathway and Its Role in Cholangiocarcinoma Metastasis. Journal of clinical medicine 2022 — PMC9267148.
- CIAPIN1 nuclear accumulation predicts poor clinical outcome in epithelial ovarian cancer. World journal of surgical oncology 2012 — PMC3502349.
- Adenovirus-delivered CIAPIN1 small interfering RNA inhibits HCC growth in vitro and in vivo. Carcinogenesis 2008 — PMC2516489.
- PubMed PMID:11230166 — UniProt-cited evidence.
- PubMed PMID:11483580 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for CIAPIN1 (Anamorsin). Expected localisation Cytoplasmic; nuclear in some tissues (HPA tissue IHC). Antigen retrieval: Tris-EDTA pH 9.0 HIER, 95–98 °C, 20 min (rule: nuclear antigen). Validated IHC antibody: A04333 at 1:50-1:200 (the product's own IHC validation image is shown). Controls (HPA): positive Testis (Pachytene spermatocytes); negative Adipose tissue. IHC workflow; specimen preparation unspecified. Protocol steps: 1. Sample: Paraffin-embedded human liver injury tissue; fixative not specified (datasheet A04333); 2. Fixation: Image fixative and duration unreported (datasheet A04333); 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-induced epitope retrieval in Tris-EDTA buffer, pH 9.0 , 20 min at 95–98 °C…; 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-CIAPIN1, 1:50-1:200 (datasheet A04333); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep fixation consistent across paraffin sections. (standard IHC practice; not target-specific); Caveat: Antibody staining and RNA show medium consistency (HPA tissue IHC); Isoform / epitope: 3 isoforms; epitope coverage is unspecified (UniProt); Regulation: Tissue abundance varies; testis is highest (HPA tissue IHC). Evidence: UniProt Q6FI81, HPA tissue IHC · Enhanced, HPA ICC-IF · Mainly localized to the mitochondria. In addition localized to the…, Datasheet A04333, PMC6481027, PMC9267148. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/CIAPIN1/immunohistochemistry-ihc-guide.