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CIAPIN1 / Anamorsin · IHC design guide
Design Immunohistochemistry for CIAPIN1
Plan CIAPIN1 paraffin-section IHC around cytoplasmic staining that can include nuclear signal (HPA tissue IHC). This guide highlights high staining in testis pachytene spermatocytes (HPA tissue IHC) and the catalog antibody’s 1:50–1:200 IHC dilution range (datasheet).
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 staining, sometimes with nuclear staining (HPA tissue IHC) | |
| Staining pattern | High in testis pachytene spermatocytes; mainly cytoplasmic (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 | Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A04333) | |
| Caveat | Marrow hematopoietic cells may be unstained (HPA tissue IHC) | |
| Regulation | Cell-type staining levels vary across tissues (HPA tissue IHC) | |
| Isoform / epitope | 3 isoforms; check epitope coverage across variants (UniProt) |
Section 1
Recommended CIAPIN1 IHC & IF Protocols
The catalog antibody’s IHC-P protocol is accompanied by 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). Citrate pH 6.0 is a published alternative (PMC3502349).
Section 2
What Is the Expected CIAPIN1 Staining Pattern?
CIAPIN1 staining should be mainly cytoplasmic in tissue IHC, with nuclear staining in some tissues (HPA tissue IHC). Expect the strongest reported signal in testicular pachytene spermatocytes and medium signal in several glandular cell populations (HPA tissue IHC). UniProt also lists cytoplasmic, nuclear and mitochondrial intermembrane space locations; CIAPIN1 has no transmembrane segment (UniProt Q6FI81). HPA rates tissue IHC reliability Enhanced, with medium consistency between staining and RNA expression (HPA tissue IHC).
| Cytoplasmic staining in pachytene spermatocytes, with some nuclear signal. | This fits the high testis signal and the possibility of combined cytoplasmic and nuclear staining (HPA tissue IHC). Score the identified cells and compartments separately; a positive testis section is a useful comparison when judging a weak sample (general IHC practice). |
| Moderate cytoplasmic staining in appendix, duodenum, gallbladder, rectum, small intestine or stomach glandular cells, or ovarian follicle cells. | These cell populations have medium reported staining (HPA tissue IHC). Compare cells within the same section before calling a tissue globally positive; HPA reports cell specific observations, and staining intensity need not be uniform across a section (HPA tissue IHC; general IHC practice). |
| Predominantly cell surface, extracellular or acellular deposits instead of cellular cytoplasmic staining. | That distribution is discordant with reported tissue IHC and the absence of a transmembrane segment (HPA tissue IHC; UniProt Q6FI81 topology). Treat it as a possible artefact and review morphology and controls before assigning CIAPIN1 positivity (general IHC practice). |
| Strong signal in a cell population reported as not detected, or broad staining outside identifiable cells. | For example, HPA reports adipocytes and bone marrow hematopoietic cells as not detected (HPA tissue IHC). Investigate antibody cross-reactivity, endogenous detection activity or nonspecific background (general IHC practice). “Not detected” is an observed IHC result, not proof that expression is biologically absent (HPA tissue IHC). |
| No staining in testicular pachytene spermatocytes while the sample and counterstain remain interpretable. | This conflicts with HPA’s high staining in those cells (HPA tissue IHC). Check section quality, antigen retrieval, antibody and detection performance using appropriate controls (general IHC practice). One failed positive section does not establish that the study samples lack CIAPIN1 (general IHC practice). |
💡Expected CIAPIN1 appearanceCall a section positive when identifiable testicular pachytene spermatocytes show strong cellular, mainly cytoplasmic staining, with possible nuclear signal; broad acellular deposits or dominant cell surface staining warrant investigation (HPA tissue IHC; UniProt Q6FI81 topology; general IHC practice).
| Tissue and cell selection | HPA reports high staining in testicular pachytene spermatocytes, medium staining in several glandular populations, and low staining in pancreatic exocrine glandular cells (HPA tissue IHC). Choose comparison cells by their reported IHC level; UniProt’s high pancreatic tissue expression does not specify an IHC intensity for every pancreatic cell (UniProt Q6FI81; HPA tissue IHC). |
| Compartment and resolution | HPA tissue IHC is mainly cytoplasmic and sometimes nuclear; UniProt also lists the mitochondrial intermembrane space (HPA tissue IHC; UniProt Q6FI81). A chromogenic cytoplasmic result need not resolve individual mitochondria (general IHC practice). The absence of a transmembrane segment gives no basis for requiring membrane staining (UniProt Q6FI81 topology). |
| Isoforms and modified residues | UniProt lists three isoforms and several phosphorylated residues (UniProt Q6FI81). The supplied sources do not map the IHC antibodies’ epitopes, so these annotations cannot predict which isoform is detected or whether phosphorylation changes staining. Interpret an unexpected pattern against tissue controls rather than assigning it to an isoform or modification (general IHC practice). |
| Evidence and antibody agreement | HPA labels tissue IHC Enhanced and reports medium consistency with RNA data; both listed antibodies have Enhanced IHC validation (HPA tissue IHC; HPA antibodies). This supports the reported pattern without making every stained structure specific. Inspect control sections when a result conflicts with the expected cell type or compartment (general IHC practice). |
| IF/ICC Q&A: Must mitochondrial puncta appear in chromogenic IHC? | No. HPA’s ICC-IF summary reports supported mitochondrial localisation with additional nucleoplasmic localisation, while its tissue IHC profile describes mainly cytoplasmic staining, sometimes with nuclear staining (HPA subcellular ICC-IF; HPA tissue IHC). Judge the IHC section against its tissue IHC pattern. |
| Situation | Likely cause | Next action |
|---|---|---|
| The testis positive comparison is blank. | The result conflicts with high staining reported in pachytene spermatocytes; section or assay performance may be responsible (HPA tissue IHC; general IHC practice). | Confirm the target cells are present, then review retrieval, antibody incubation, chromogenic detection and control performance before interpreting study sections (general IHC practice). |
| All cells show a diffuse brown haze. | Nonspecific staining or endogenous detection activity can obscure cellular localisation (general IHC practice). | Inspect a no-primary control, review blocking and washing, and adjust the staining workflow until identifiable cellular signal can be distinguished from background (general IHC practice). |
| Only nuclei stain strongly across a tissue section. | Some nuclear signal is compatible with HPA, but a uniformly nuclear-only pattern differs from its mainly cytoplasmic tissue profile (HPA tissue IHC). | Compare with cytoplasmic staining in an appropriate positive section and review background and antibody controls before scoring the nuclei as specific (HPA tissue IHC; general IHC practice). |
| Adipocytes or bone marrow hematopoietic cells appear strongly positive. | HPA reports these cell populations as not detected by tissue IHC, making strong staining discordant with its observations (HPA tissue IHC). | Check cell identification and a no-primary control; assess cross-reactivity or endogenous detection activity before interpreting the signal as CIAPIN1 (general IHC practice). |
| Pancreatic exocrine glandular cells stain weakly. | HPA reports low IHC staining in these cells, although UniProt describes high expression in pancreas at the tissue level (HPA tissue IHC; UniProt Q6FI81). | Use the cell specific IHC observation when judging the section; compare a known stronger IHC population if assay sensitivity remains uncertain (HPA tissue IHC; general IHC practice). |
| Signal pools at section edges or outside cells. | Edge or acellular deposits do not match the cellular cytoplasmic IHC profile (HPA tissue IHC; general IHC practice). | Review section morphology, washing and chromogen development, then score only interpretable cellular staining alongside appropriate controls (general IHC practice). |
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); use adipose tissue adipocytes as the biological negative (HPA: Not detected). On the testis slide, cells without visible target staining should remain at background, but the supplied HPA rows do not identify a specific testicular cell type as an internal negative (HPA: testis row).
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 no-primary (secondary-only), a host- and clonality-matched isotype control, and CIAPIN1 knockout tissue or a validated immunizing-peptide block where available (standard IHC practice). For chromogenic testis IHC, quench endogenous peroxidase and check the controls for residual background (standard IHC practice).
⚠️Feasibility: No matched source reports a CIAPIN1-specific fixation window, retrieval dependency, or whether frozen-section staining or IF is easier; optimize antigen retrieval for the IHC-validated antibody (supplied target/application evidence; standard IHC practice). The selected A04333 paraffin-section caption does not report a fixative, so fixation cannot be established from it (selected A04333 caption). Endogenous peroxidase can cause background in chromogenic testis sections (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 by checking retrieval, compartment, controls and cell-specific scoring before interpreting signal intensity.
How should I retrieve CIAPIN1 in paraffin sections when nuclear staining is weak?
Start with Tris-EDTA at pH 9.0 for heat-induced retrieval at 95–98 °C for 20 min (page retrieval rule). Let sections cool in buffer, then compare nuclear and cytoplasmic signal in matched sections stained with the same antibody dilution (standard IHC practice; HPA: cytoplasmic expression sometimes combined with nuclear expression). If staining remains weak, test a milder retrieval condition alongside the specified condition while holding detection constant (standard IHC practice). Record tissue preservation and background as well as signal, because aggressive heating can damage morphology and complicate compartment scoring (standard IHC practice).
Could fixation explain weak CIAPIN1 staining in my paraffin sections?
CIAPIN1-specific sensitivity to fixation is unknown from the supplied evidence; the catalog image identifies paraffin-embedded human liver injury but does not state a fixative (A04333 caption). Record the actual fixative and fixation interval for each specimen, and compare sections processed together before attributing intensity differences to biology (standard IHC practice). Check whether morphology and a separate process-control stain are preserved, since poor processing can affect interpretation of chromogenic signal (standard IHC practice). Keep retrieval, antibody dilution and detection timing constant during that comparison; a staining difference alone cannot establish a CIAPIN1-specific fixation effect (standard IHC practice).
Should CIAPIN1 stain nuclei, cytoplasm or mitochondria in tissue IHC?
Expect predominantly cytoplasmic tissue staining, with nuclear staining in some tissues (HPA: tissue IHC profile). UniProt also lists cytoplasm, nucleus and mitochondrial intermembrane space, while cellular IF supports mitochondria as the main location and nucleoplasm as an additional location (UniProt Q6FI81 subcellular; HPA: subcellular). In chromogenic sections, score nuclear and cytoplasmic staining separately, because a diffuse cytoplasmic deposit cannot establish mitochondrial localization by itself (standard IHC interpretation). Compare the pattern with morphology and matched controls before treating a newly dominant compartment as biological redistribution (standard IHC practice; HPA: tissue IHC profile).
Could isoforms or epitope masking explain inconsistent CIAPIN1 staining?
CIAPIN1 has 3 annotated isoforms, and the supplied evidence does not identify the catalog antibody’s epitope or isoform coverage (UniProt Q6FI81 isoforms; supplied antibody evidence). It also lists phosphorylation at residues 136, 177, 182, 183 and 215, without showing that these modifications alter this antibody’s binding (UniProt Q6FI81 modified residues). Compare retrieval conditions on adjacent sections and keep antibody dilution and detection constant when investigating epitope accessibility (standard IHC practice). Do not assign a staining difference to a particular isoform or phosphorylation event without epitope mapping and an independent validation method (standard IHC interpretation).
How can IF help resolve ambiguous CIAPIN1 compartment staining?
Use IF as a separate localization check and multiplex CIAPIN1 with a marker for the cell population being evaluated, such as pachytene spermatocytes when examining testis (HPA: high in pachytene spermatocytes). Choose a fluorophore channel after checking the specimen’s autofluorescence, and include single-color controls when interpreting overlap (standard IF practice). Optimize permeabilisation for cytoplasmic and nuclear access; assessment of the mitochondrial intermembrane space may require additional membrane access, so verify organelle morphology after treatment (UniProt Q6FI81 subcellular; standard IF practice). Compare any punctate mitochondrial pattern with an organelle marker rather than inferring mitochondria from puncta alone (HPA: mitochondrial localization supported; standard IF interpretation).
What should I check when CIAPIN1 staining is widespread or granular?
First separate expected broad expression from nonspecific deposit: CIAPIN1 is ubiquitously expressed, and tissue IHC shows cytoplasmic staining in most tissues (UniProt Q6FI81 tissue specificity; HPA: tissue IHC profile). Run a no-primary control, review the peroxidase block and inspect whether chromogen appears over tissue edges or damaged areas (standard chromogenic IHC practice). Titrate the antibody and shorten chromogen development in matched sections if background obscures cellular boundaries (standard IHC practice). Use the same imaging and counterstain settings when comparing sections, because apparent granular intensity can shift with detection conditions (standard IHC practice).
How should I quantify CIAPIN1 across paraffin tissue sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring, because CIAPIN1 tissue staining is usually cytoplasmic and can include nuclear signal (HPA: tissue IHC profile). For each compartment, report the percentage of positive cells and an H-score using intensity categories 0–3; reserve positive-cell density per mm² for questions about cell abundance (standard IHC scoring practice). Normalize counts to the number of eligible cells or sampled viable tissue area, using comparable section areas and imaging settings (standard IHC practice). Keep cytoplasmic and nuclear scores separate and document excluded damaged regions so changes in composition do not masquerade as changes in staining (standard IHC practice).
How do I distinguish genuine CIAPIN1 signal from chromogenic artefact?
A plausible positive pattern includes cytoplasmic staining in many tissues, sometimes with nuclear staining; testis pachytene spermatocytes are reported as high (HPA: tissue IHC profile; HPA: high in pachytene spermatocytes). Treat a signal confined to tissue edges, necrotic regions or a no-primary control as suspect, and check the peroxidase block when diffuse chromogen persists (standard chromogenic IHC practice). A dominant compartment that conflicts with the expected pattern needs confirmation with an independent method before interpretation (HPA: tissue IHC profile; standard IHC interpretation). The catalog image demonstrates staining in paraffin-embedded human liver injury, but its caption does not establish fixation conditions or specificity in every cell type (A04333 caption).
Boster reagents
Best CIAPIN1 / Anamorsin IHC Antibodies
A04333 is listed for human and mouse IHC and IF (catalog: applications, reactivity). Its IHC figure shows a paraffin-embedded human liver injury specimen (A04333 image caption); no IF figure is supplied (catalog: IF image alts).
Real IHC data
Anti-Anamorsin CIAPIN1 Antibody
Cat # A04333
A04333 lists IHC and IF applications and human and mouse reactivity (catalog: applications, reactivity). Its IHC figure depicts paraffin-embedded human liver injury (A04333 image caption); the catalog supplies no IF figure (catalog: IF image alts).
Which to pick: For tissue IHC, choose A04333: its own figure shows a paraffin-embedded human liver injury specimen, but the fixative is unreported (A04333 image caption). For IF/ICC, A04333 lists IF at 1:50–1:200; no IF figure or ICC validation is supplied (catalog: applications, IF dilution, IF image alts). For cross-species work, A04333 lists human and mouse reactivity and a rabbit host; clonality is unreported (catalog: reactivity, host, clone).
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 staining, sometimes with nuclear staining (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: Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A04333); Caveat: Marrow hematopoietic cells may be unstained (HPA tissue IHC); Isoform / epitope: 3 isoforms; check epitope coverage across variants (UniProt); Regulation: Cell-type staining levels vary across tissues (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.