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CCDC88A / Girdin · IHC design guide
Design Immunohistochemistry for CCDC88A
Plan paraffin IHC for CCDC88A using cytoplasmic staining in lymphatic organs as a reference (HPA tissue IHC). Use testis pachytene spermatocytes and adipose adipocytes as positive and negative controls (HPA tissue IHC), and titrate the IHC-validated antibody from 1:100–1:300 (datasheet: A03282-1).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
CCDC88A 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 in lymphatic organs (HPA tissue IHC) | |
| Staining pattern | Cytoplasmic staining in lymphatic germinal center cells (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane 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 conditions consistent across sections. (standard IHC practice; not target-specific) | |
| Caveat | Lung macrophages may complicate parenchymal scoring (HPA tissue IHC) | |
| Regulation | EGF promotes membrane localization (UniProt) | |
| Isoform / epitope | 5 isoforms; no transmembrane segment; verify epitope coverage (UniProt) |
Section 1
Recommended CCDC88A IHC & IF Protocols
The catalog antibody has an IHC-P protocol (datasheet); the published protocols below stained pancreatic cancer and hepatocellular carcinoma tissue sections (PMC5139074; PMC9043556).
| Sample | Paraffin-embedded human brain tissue; fixative not specified (datasheet A03282-1) |
| Fixation | Image fixative and duration unreported (datasheet A03282-1); 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 citrate buffer, pH 6.0, 20 min at 95–98 °C (standard rule: cytoplasmic / membrane 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-CCDC88A, 1:100 - 1:300 (datasheet A03282-1) |
| 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 | CCDC88A-positive staining in pachytene spermatocytes of testis (HPA tissue IHC: High). HPA tissue profile: Abundant cytoplasmic expression in lymphatic organs. No signal in the no-primary control. |
💡Decision noteStart with citrate pH 6.0 at 95–98 °C for 20 min (page retrieval); the pancreatic cancer study used an autoclave at 108 °C for 15 min (PMC5139074).
Section 2
What Is the Expected CCDC88A Staining Pattern?
CCDC88A (Girdin) is found in the cytosol, cytoskeleton, vesicles and cell membrane; it has no transmembrane segment (UniProt Q3V6T2). In paraffin-section IHC, expect predominantly cytoplasmic staining, particularly in pachytene spermatocytes, lung macrophages and lymphoid cells (HPA tissue IHC). HPA rates tissue IHC reliability as Enhanced, with medium consistency between staining and RNA expression (HPA tissue IHC).
| Strong cytoplasmic staining in testicular pachytene spermatocytes, with distinguishable cell boundaries. | This matches the reported High signal in those cells (HPA tissue IHC). Compare staining with the counterstain and tissue structure before scoring it (general IHC practice). |
| Moderate cytoplasmic staining in lung macrophages or germinal-center cells of lymph node or tonsil. | These cell populations are reported at Medium intensity (HPA tissue IHC). Score the identified cells, rather than treating the whole section as uniformly positive (general IHC practice). |
| Predominantly nuclear staining in paraffin sections, without a convincing cytoplasmic pattern. | Treat isolated nuclear IHC signal cautiously: UniProt lists cytoplasmic and membrane locations, while HPA calls nucleoplasmic ICC-IF localization uncertain (UniProt Q3V6T2; HPA subcellular). Check controls before assigning a nuclear IHC pattern (general IHC practice). |
| Strong staining in adipocytes or bronchial respiratory epithelium while expected positive cells are weak. | HPA reports those cell types as Not detected (HPA tissue IHC). Assess possible antibody cross-reactivity or endogenous chromogenic activity with appropriate controls (general IHC practice). |
| Diffuse color across cells and empty spaces, or no signal in otherwise intact testicular pachytene spermatocytes. | Diffuse color limits cellular interpretation; absent signal conflicts with the reported High testis pattern (HPA tissue IHC). Review background and positive controls before calling the sample negative (general IHC practice). |
💡Expected CCDC88A appearanceA convincing positive is predominantly cytoplasmic, High in testicular pachytene spermatocytes or Medium in the specified macrophage and lymphoid populations (HPA tissue IHC); widespread featureless color is suspect background (general IHC practice).
| Cell population and tissue | Signal varies by cell type: pachytene spermatocytes are High; lung macrophages and germinal-center cells are Medium; adipocytes are Not detected (HPA tissue IHC). |
| Location and stimulation | UniProt places unstimulated CCDC88A in cytosol and reports EGF-promoted membrane localization. Membrane accentuation can therefore have a biological basis, but tissue IHC alone does not establish stimulation state (UniProt Q3V6T2). |
| Antibody validation | HPA rates HPA038102 IHC as Enhanced and HPA038101 as Supported. Validation supports comparison with reported tissue patterns; it does not guarantee every section will reproduce them (HPA antibodies; general IHC practice). |
| Isoforms and epitope | UniProt lists 5 isoforms (UniProt Q3V6T2). The supplied evidence gives no antibody epitope, so it cannot establish which isoforms a particular IHC stain detects. |
| Protein topology and processing | UniProt lists no transmembrane segment or signal peptide and describes one chain spanning residues 1–1871 (UniProt Q3V6T2). Do not infer a secreted or shed-protein staining pattern from this record. |
| Situation | Likely cause | Next action |
|---|---|---|
| No signal in testicular pachytene spermatocytes. | The expected cell population is reported High, so a negative result needs technical review before interpretation (HPA tissue IHC; general IHC practice). | Confirm tissue identity and a working positive control; review retrieval, antibody dilution and chromogenic detection against the chosen IHC protocol (general IHC practice). |
| Strong color in adipocytes or bronchial respiratory epithelium. | Both cell types are reported Not detected; staining there may reflect nonspecific binding or endogenous detection activity (HPA tissue IHC; general IHC practice). | Review cell identity and a no-primary control; check blocking and endogenous-enzyme controls appropriate to the detection system (general IHC practice). |
| Color appears evenly across the section, including spaces between cells. | A noncellular distribution is consistent with background and does not identify a CCDC88A-positive cell population (general IHC practice). | Inspect no-primary and tissue controls; review blocking, washing, antibody dilution and chromogen development (general IHC practice). |
| Only nuclei appear positive in IHC. | HPA labels nucleoplasmic ICC-IF localization uncertain; UniProt also lists several cytoplasmic locations (HPA subcellular; UniProt Q3V6T2). | Check controls and the cytoplasmic compartment before interpreting nuclear-only IHC as target-specific (general IHC practice). |
| Membrane-edge staining varies among otherwise similar cells. | CCDC88A can move toward the membrane after EGF stimulation, but this slide alone cannot identify the stimulus (UniProt Q3V6T2). | Score cytoplasmic and membrane-associated staining separately, and compare like cell types on the same section (general IHC practice). |
| Q: Does ICC-IF need the same expected pattern as paraffin-section IHC? | HPA ICC-IF reports supported centrosome, basal-body and plasma-membrane locations, but uncertain nucleoplasm localization (HPA subcellular). | A: Interpret ICC-IF compartments using its own controls and guide; do not transfer those localization calls directly to chromogenic tissue IHC (general IHC practice). |
Experiment evidence
IHC & IF images showing CCDC88A staining
A03282-1 · IHCImmunohistochemistry analysis of paraffin-embedded human brain tissue, using Girdin Antibody. The picture on the right is blocked with the s
P03282 · IHCImmunohistochemistry analysis of paraffin-embedded human breast carcinoma, using Girdin (Phospho-Ser1417) Antibody. The picture on the right
A03282-2 · IFImmunofluorescence of Girdin in human brain tissue with Girdin antibody at 20 μg/mL.
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000115355-CCDC88A/tissue/cerebral+cortex#imid_9664102
HPA · EsophagusHuman Protein Atlas IHC, Esophagus. Source: https://www.proteinatlas.org/ENSG00000115355-CCDC88A/tissue/esophagus#imid_9664074
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000115355-CCDC88A/tissue/colon#imid_9657676
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000115355-CCDC88A/tissue/liver#imid_9657693
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA038102; green = CCDC88A, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA038102; green = CCDC88A, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for CCDC88A IHC & IF
🧪Run testis first and require staining in pachytene spermatocytes (High; HPA: testis). Use adipose tissue adipocytes as the negative comparator (Not detected; HPA: adipose tissue); on the testis slide, assess morphologically distinct cells without signal as internal negative comparators without assuming all other testicular cells are negative (HPA: testis).
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 CCDC88A in A-431, U-251MG, U2OS, RPTEC/TERT1, hTERT-RPE1 (serum starved), with annotated localisation: Nucleoplasm (uncertain), Centrosome (supported), Basal body (supported) (HPA subcellular).
Technical controls: Include no-primary (secondary-only) and host-species- and immunoglobulin-class-matched isotype controls (standard IHC practice). Use a CCDC88A knockout specimen if available, or a peptide-block control as shown for the catalog antibody (selected IHC caption: A03282-1); quench endogenous peroxidase and check residual background in testis before interpreting chromogenic signal (standard IHC practice).
⚠️Feasibility: A target-specific fixation window and the effect of fixation on staining are unreported; the paraffin-section caption for A03282-1 does not state a fixative (selected IHC caption: A03282-1). Antigen-retrieval dependence is unreported, and the supplied evidence does not establish whether frozen sections or IF/ICC are easier than paraffin IHC (selected IHC caption: A03282-1; HPA: ICC-IF images). In testis, densely packed germ cells can complicate identification of pachytene spermatocytes, so assess staining with tissue morphology and a counterstain (standard IHC practice; HPA: testis).
HPA tissue IHC evidence for CCDC88A
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 | Lymphoid tissue | Medium | Protein (IHC) | HPA → |
| Lung | Macrophages | Medium | Protein (IHC) | HPA → |
| Lymph node | Germinal center cells | Medium | Protein (IHC) | HPA → |
| Tonsil | Germinal center cells | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Adrenal gland | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Breast | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | Not detected | Protein (IHC) | HPA → |
| Caudate | Glial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced CCDC88A IHC Tips
Troubleshoot CCDC88A (Girdin) staining in paraffin-section chromogenic IHC using matched controls and compartment-aware interpretation.
How should I adjust retrieval when Girdin staining is weak?
For paraffin-section IHC, start with citrate pH 6.0 HIER at 95–98 °C for 20 min (page retrieval rule). Run a known positive section alongside the test section, then compare cytoplasmic signal at matched antibody dilution and development time (HPA: cytoplasmic tissue expression; standard IHC practice). If staining remains weak, test a more alkaline retrieval buffer on a serial section while keeping detection conditions fixed (standard IHC practice). Excess retrieval can damage tissue morphology or increase diffuse staining, so inspect both signal and section integrity (standard IHC practice). Do not interpret a negative result until the positive control stains appropriately (standard IHC practice).
Could fixation explain a negative Girdin IHC result?
Target-specific fixation sensitivity for CCDC88A is unknown from the supplied evidence; the selected antibody caption describes paraffin-embedded human brain tissue but does not state its fixative (A03282-1 caption). Record the actual fixative and fixation duration for each specimen before comparing staining across cases (standard IHC practice). If fixation varies, stain serial sections in the same run with identical pH 6.0 retrieval and antibody conditions (page retrieval rule; standard IHC practice). Examine morphology and a known positive control before attributing weak signal to fixation (standard IHC practice). Tissue staining patterns and protein topology do not establish a fixation effect for this antibody (HPA: tissue IHC profile; UniProt Q3V6T2 topology).
Which staining compartments are credible for Girdin in tissue sections?
Expect substantial cytoplasmic staining in suitable tissue cells; membrane-associated and focal centrosomal or basal-body signal can also be biologically plausible (HPA: cytoplasmic tissue profile; UniProt Q3V6T2 subcellular; HPA: supported centrosome, basal body and plasma membrane). Girdin has no transmembrane segment, so a membrane pattern alone does not imply an integral membrane protein (UniProt Q3V6T2 topology). Its reported cytosol-to-membrane redistribution follows EGF stimulation, which should not be assumed for an unstimulated section (UniProt Q3V6T2 subcellular). Compare the cellular pattern with adjacent morphology at the same pH 6.0 retrieval setting (page retrieval rule; standard IHC practice). Diffuse nuclear-only chromogen warrants control review because HPA calls nucleoplasmic localisation uncertain (HPA: subcellular localisation).
Can this IHC antibody distinguish Girdin isoforms or phosphorylation states?
CCDC88A has 5 reported isoforms, but the supplied antibody evidence does not identify its epitope or demonstrate isoform selectivity (UniProt Q3V6T2 isoforms; A03282-1 caption). The protein also has documented modified residues, including phosphoserines at 233 and 237 (UniProt Q3V6T2 modified residues). Treat routine chromogenic signal as antibody-reactive Girdin, without assigning an isoform or phosphorylation state (UniProt Q3V6T2 isoforms and modified residues; standard IHC interpretation). The caption’s peptide-blocked image supports a competition check for that antibody, but does not establish which isoform contributes to tissue staining (A03282-1 caption). Resolve a specific epitope question with documented antibody mapping or an independent, appropriately validated reagent (standard IHC practice).
How should an IF follow-up test Girdin localisation?
Use the separate IF/ICC workflow for fluorescence imaging, while retaining chromogenic IHC as this page’s primary application (application specification). Multiplex Girdin with a marker for the expected cell population, such as a macrophage marker when examining lung macrophages, and assess overlap cell by cell (HPA: medium staining in lung macrophages; standard IF practice). Select a fluorophore channel after checking unstained tissue autofluorescence, and include single-stain controls for channel bleed-through (standard IF practice). If the antibody epitope is intracellular, use validated permeabilisation; if it is extracellular, avoid assuming that intracellular access is needed (standard IF practice). Girdin lacks a transmembrane segment, and the supplied evidence does not locate this antibody’s epitope (UniProt Q3V6T2 topology; A03282-1 caption).
How can I reduce widespread brown background without losing Girdin signal?
First compare the stained section with a no-primary control and inspect whether brown signal follows cells, section edges or endogenous pigment (standard chromogenic IHC practice). Block endogenous peroxidase before chromogen development and titrate the IHC-validated antibody against a positive section (standard chromogenic IHC practice). If background persists, increase wash stringency or shorten chromogen development while holding pH 6.0 retrieval constant (page retrieval rule; standard IHC practice). Girdin can be cytoplasmic and membrane-associated, so broad intracellular staining alone is not proof of nonspecific binding (UniProt Q3V6T2 subcellular; HPA: cytoplasmic tissue profile). The peptide-blocked companion image offers an antibody-specific comparison for the selected product (A03282-1 caption).
What should I score when comparing Girdin staining across sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring, then use an H-score based on intensity and percentage of positive cells for comparable tissue regions (HPA: cytoplasmic tissue profile; standard IHC scoring practice). Alternatively, report percentage positive cells or positive-cell density per mm² when intensity is unreliable (standard IHC scoring practice). Normalise each result to the number of eligible cells or the measured viable tissue area, and exclude folds, section edges and necrosis consistently (standard IHC scoring practice). Keep retrieval, antibody dilution, counterstain and chromogen development matched across slides (page retrieval rule; standard IHC practice). Report cytoplasmic and membrane-associated scores separately if both are measured (UniProt Q3V6T2 subcellular; standard IHC scoring practice).
How do I distinguish true Girdin staining from artefact?
A credible result follows identifiable cells and a plausible cytoplasmic, membrane-associated or focal centrosomal pattern, with an appropriate positive control (HPA: cytoplasmic tissue profile; UniProt Q3V6T2 subcellular; standard IHC practice). Check the cell identity: HPA reports medium staining in lung macrophages and no detection in bronchial respiratory epithelial cells (HPA: lung macrophages; HPA: bronchus respiratory epithelial cells). Be cautious with nuclear-only signal because HPA labels nucleoplasmic localisation uncertain (HPA: subcellular localisation). Edge accentuation, necrotic deposits and staining in a no-primary control favour processing or detection artefact; check endogenous peroxidase blocking when using chromogen (standard chromogenic IHC practice). Compare suspected signal with the selected antibody’s peptide-blocked companion image where relevant (A03282-1 caption).
Boster reagents
Best CCDC88A / Girdin IHC Antibodies
The catalog shows CCDC88A IHC in human brain and breast carcinoma sections and IF in human brain tissue (image captions). Listed reactivity covers human and mouse, plus rat for A03282-2 (catalog).
Real IHC data
Anti-Girdin CCDC88A Antibody
Cat # A03282-1
Real IHC data
Anti-Phospho Girdin (Ser1417) CCDC88A Antibody
Cat # P03282
Real IF data
Anti-Girdin CCDC88A Antibody
Cat # A03282-2
A03282-1 has an IHC image of paraffin-embedded human brain; P03282 targets phospho-Ser1417 and has an IHC image of paraffin-embedded human breast carcinoma (catalog image captions). A03282-2 has an IF image of human brain tissue (catalog image caption).
Which to pick: For tissue IHC, choose A03282-1 for total Girdin or P03282 for phospho-Ser1417; both list IHC at 1:100–1:300, with paraffin-section images for A03282-1 in human brain and P03282 in human breast carcinoma (catalog applications, dilution fields, and each SKU’s image caption). For IF/ICC, A03282-1 and P03282 list both applications; A03282-2 lists IF and shows human brain IF, but does not list ICC or IHC (catalog applications and A03282-2 image caption). For broader listed species reactivity, A03282-2 covers human, mouse, and rat, although its pictured IF evidence is human only (catalog reactivity and A03282-2 image caption); neither IHC caption reports a fixative (A03282-1 and P03282 image captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q3V6T2 (GRDN_HUMAN, Girdin).
- Human Protein Atlas. CCDC88A tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. CCDC88A subcellular location (ICC-IF): Mainly localized to the nucleoplasm, centrosome and basal body. In addition localized to the plasma membrane..
- Human Protein Atlas. CCDC88A antibody validation summary (2 antibodies).
- CCDC88A, a prognostic factor for human pancreatic cancers, promotes the motility and invasiveness of pancreatic cancer cells. Journal of experimental & clinical cancer research : CR 2016 — PMC5139074.
- CCDC88A Post-Transcriptionally Regulates VEGF via miR-101 and Subsequently Regulates Hepatocellular Carcinoma. Frontiers in immunology 2022 — PMC9043556.
- Efficient delivery of small interfering RNAs targeting particular mRNAs into pancreatic cancer cells inhibits invasiveness and metastasis of pancreatic tumors. Oncotarget 2019 — PMC6499602.
- PubMed PMID:16139227 — UniProt-cited evidence.
- PubMed PMID:15882442 — UniProt-cited evidence.
- PubMed PMID:15815621 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for CCDC88A (Girdin). Expected localisation Cytoplasmic staining in lymphatic organs (HPA tissue IHC). Antigen retrieval: Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen). Validated IHC antibody: A03282-1 at 1:100 - 1:300 (the product's own IHC validation image is shown). Controls (HPA): positive Testis (Pachytene spermatocytes); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human brain tissue; fixative not specified (datasheet A03282-1); 2. Fixation: Image fixative and duration unreported (datasheet A03282-1); 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-induced epitope retrieval in citrate buffer, pH 6.0 , 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-CCDC88A, 1:100 - 1:300 (datasheet A03282-1); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep fixation conditions consistent across sections. (standard IHC practice; not target-specific); Caveat: Lung macrophages may complicate parenchymal scoring (HPA tissue IHC); Isoform / epitope: 5 isoforms; no transmembrane segment; verify epitope coverage (UniProt); Regulation: EGF promotes membrane localization (UniProt). Evidence: UniProt Q3V6T2, HPA tissue IHC · Enhanced, HPA ICC-IF · Mainly localized to the nucleoplasm, centrosome and basal body. In…, Datasheet A03282-1, PMC5139074, PMC9043556. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/CCDC88A/immunohistochemistry-ihc-guide.