COPS5 / COP9 signalosome complex subunit 5 · IHC design guide

Design Immunohistochemistry for COPS5

Plan COPS5 chromogenic IHC on paraffin sections using an IHC-validated antibody and high-staining tissue controls (HPA tissue IHC). Assess nuclear and cytoplasmic staining separately, and start the catalog antibody at 2 μg/mL (HPA tissue IHC; datasheet M01849-2).

Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
Immunohistochemistry protocol sheet for COPS5 (IHC for COPS5): expected localisation Nuclear and cytoplasmic tissue staining (HPA tissue IHC), antibody M01849-2, validated IHC image, and IHC protocol steps
Printable COPS5 IHC protocol sheet — expected localisation Nuclear and cytoplasmic tissue staining (HPA tissue IHC), antibody M01849-2, controls and protocol steps. Open the full COPS5 IHC guide →

COPS5 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 Nuclear and cytoplasmic tissue staining (HPA tissue IHC)
Staining pattern Nuclear and cytoplasmic staining in cells across tissues (HPA tissue IHC)
Antigen retrieval EDTA pH 8.0 HIER, heat-mediated (datasheet M01849-2)
Positive control ⓘ Adipose tissue+4 more · see all
Negative control ⓘ None in HPA (detected in all 45 tissues); use no-primary + isotype controls
Important caveats
Reasons your staining may differ from the expected pattern.
Fixation Keep paraffin-section fixation consistent across specimens. (selected-SKU IHC image M01849-2)
Caveat IFIT3 can diminish nuclear localization (UniProt)
Regulation No intensity regulator annotated (UniProt)
Isoform / epitope No isoforms annotated; chain spans residues 2–334 (UniProt)
Section 1

Recommended COPS5 IHC & IF Protocols

Compare the catalog antibody’s EDTA pH 8.0 retrieval protocol (datasheet M01849-2) with published COPS5 IHC protocols for paraffin sections (PMC4932188; PMC5722579).

Recommended immunohistochemistry (IHC-P) protocol parameters
SampleParaffin-embedded human esophageal squamous carcinoma tissue; fixative not specified (datasheet M01849-2)
FixationImage fixative and duration unreported (datasheet M01849-2); verify before use.
Sectioning4–5 µm sections on charged slides (standard)
DeparaffinisationXylene, graded ethanol series to water (standard)
Antigen retrievalHeat retrieval: EDTA pH 8.0 (datasheet M01849-2); 20 min, 95–100 °C (standard)
Peroxidase block3% H2O2, 10 min, room temperature (standard)
Blocking10% goat serum (datasheet M01849-2)
Primary antibodyMouse monoclonal (clone 4G9) anti-COPS5, 2μg/ml (datasheet M01849-2)
Primary incubationOvernight at 4 °C (datasheet M01849-2)
DetectionStreptavidin-biotin complex (SABC), DAB chromogen (datasheet M01849-2)
CounterstainHematoxylin, blue, dehydrate and mount (standard)
Expected resultCOPS5-positive staining in adipocytes of adipose tissue (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic and nuclear expression in all tissues. No signal in the no-primary control.
💡Decision noteStart with heat-mediated EDTA pH 8.0 retrieval (datasheet M01849-2); consider citrate pH 6.0 pressure-cooker retrieval during optimization (PMC5722579).
Section 2

What Is the Expected COPS5 Staining Pattern?

COPS5 is a soluble protein found in the cytosol, nucleus and perinuclear region; it has no transmembrane segment (UniProt Q92905). In tissue sections, expect cytoplasmic and nuclear staining across cell types, including strong staining in the listed HPA high-expression cells (HPA: tissue IHC). HPA rates its tissue pattern Supported, with medium consistency between staining and RNA data (HPA: tissue IHC).

What am I looking at on my slide?
Nuclear and cytoplasmic staining in adipocytes, glandular cells or hematopoietic cells.This fits the reported tissue pattern; HPA grades these cell types High in specified tissues (HPA: tissue IHC). Assess staining within cells, while allowing nuclear-to-cytoplasmic balance to vary: both compartments are reported (UniProt Q92905; HPA: tissue IHC).
Predominantly sharp cell-surface or extracellular staining, with little intracellular signal.Treat this as a localization mismatch to investigate, since COPS5 has no transmembrane segment and its reported sites are intracellular (UniProt Q92905). Check whether the pattern follows section edges, deposits or structures outside cells before interpreting it as COPS5 (general IHC practice).
Staining is confined to an unexpected population while the expected cells are blank.A single stained cell type is hard to reconcile with HPA's cytoplasmic and nuclear expression across tissues (HPA: tissue IHC). Consider antibody cross-reactivity or endogenous detection activity; compare morphology and a detection control before assigning the signal to COPS5 (general IHC practice).
Haze or granular chromogen covers many structures without clear cell boundaries.This distribution cannot establish nuclear or cytoplasmic localization (general IHC practice). Review blocking, washes and detection controls for background, then judge whether recognizable cells retain the reported intracellular pattern (HPA: tissue IHC; general IHC practice).
No detectable signal in a section containing an HPA High cell type.A negative result there conflicts with the reported staining, but does not by itself prove COPS5 is absent (HPA: tissue IHC). Check tissue preservation, retrieval and detection performance with appropriate controls before drawing a biological conclusion (general IHC practice).
💡Expected COPS5 appearanceCall a section positive when recognizable cells show cytoplasmic and nuclear chromogen, with strong staining plausible in HPA High cell types; isolated surface or cell-free deposits are suspect rather than convincing COPS5 staining (HPA: tissue IHC; UniProt Q92905 topology; general IHC practice).
How each factor affects the staining
Tissue distributionHPA reports cytoplasmic and nuclear expression in all tissues and low tissue RNA specificity; its listed High cells include adipocytes, glandular, hematopoietic, respiratory epithelial and glial cells (HPA: tissue IHC). Use the stated cell type within its listed tissue when selecting an expected-positive area.
Location and topologyUniProt places COPS5 in cytosol, nucleus and perinuclear cytoplasm, with no transmembrane segment (UniProt Q92905). This supports intracellular interpretation; a membrane-dominant pattern needs independent scrutiny (UniProt Q92905; general IHC practice).
Evidence strengthThe tissue profile is Supported with medium staining-to-RNA consistency; HPA004845 and CAB004242 are IHC Supported (HPA: tissue IHC; HPA: antibodies). These ratings support comparison with the reported pattern, without making every individual stained structure specific.
Processing and fixation evidenceUniProt lists no signal peptide or propeptide and no glycosylation sites (UniProt Q92905). The supplied sources do not report COPS5-specific fixation sensitivity or retrieval conditions; choose and assess retrieval using general IHC controls (general IHC practice).
IF/ICC Q&A: should its image match tissue IHC exactly?No exact match is established: HPA reports enhanced nucleoplasmic localization in ICC-IF images, while tissue IHC reports nuclear and cytoplasmic staining (HPA: subcellular; HPA: tissue IHC). Interpret each result in its own specimen and detection context.
Why is my staining missing, weak or wrong?
SituationLikely causeNext action
Blank staining in an expected-positive area.A failed retrieval or detection step is possible; the sources give no COPS5-specific retrieval setting (general IHC practice).Confirm the listed HPA High cell type is present, then check a positive control and the IHC-validated antibody's documented procedure (HPA: tissue IHC; general IHC practice).
Uniform brown haze obscures cells.Nonspecific reagent binding, inadequate washing or endogenous detection activity can produce background (general IHC practice).Review blocking and wash steps; use an appropriate detection control and score only cell-associated nuclear or cytoplasmic signal (general IHC practice; HPA: tissue IHC).
Staining appears only on cell borders or outside cells.That placement disagrees with COPS5's intracellular locations and lack of a transmembrane segment (UniProt Q92905).Inspect morphology and deposits, compare a detection control, and re-evaluate intracellular staining before calling the section positive (general IHC practice).
Only one unexpected cell population stains.Cross-reactivity or endogenous detection activity is possible; HPA describes expression across tissues (HPA: tissue IHC; general IHC practice).Identify the stained cells morphologically and compare expected-positive cells and detection controls; avoid assigning specificity from color alone (HPA: tissue IHC; general IHC practice).
Nuclear staining dominates, with faint cytoplasm.Compartment balance may differ by specimen: UniProt lists both locations, and HPA ICC-IF emphasizes nucleoplasm (UniProt Q92905; HPA: subcellular).Check whether nuclear signal is cell-associated and whether cytoplasm is discernible; compare tissue IHC controls before rejecting the pattern (HPA: tissue IHC; general IHC practice).
Intensity varies between areas of one section.Cell composition or staining performance may differ across areas (general IHC practice); HPA High calls apply to named cell types in specified tissues (HPA: tissue IHC).Compare like cell types in intact areas, inspect section quality and controls, and record compartment and intensity separately (HPA: tissue IHC; general IHC practice).

Sample controls for COPS5 IHC & IF

🧪Run adipose tissue first and confirm staining in adipocytes (HPA: High in adipocytes). HPA detects COPS5 in all 45 scored tissues, so there is no validated negative tissue; no-primary and isotype controls establish background, and any cells without specific staining on the positive slide should show only counterstain or background, without being treated as validated target-negative cells (HPA: no negative rows).
Positive control tissue: Adipose tissue (Adipocytes, HPA High)
Negative control tissue: None in HPA: COPS5 is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead.
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show COPS5 in A-431, K-562, U2OS, with annotated localisation: Nucleoplasm (enhanced) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only slide; an isotype-matched mouse control IgG for the caption antibody; and COPS5-knockout material as a biological negative where available (caption: mouse anti-JAB1 M01849-2; standard IHC controls). For chromogenic detection, block endogenous peroxidase and assess endogenous biotin because the reported workflow uses streptavidin–biotin detection with DAB (caption: SABC/DAB).
⚠️Feasibility: A target-specific fixation window and fixation effect are unreported, and the selected paraffin-section caption does not state a fixative (caption: fixative not stated). The reported IHC workflow uses heat retrieval in EDTA at pH 8.0, but the evidence does not establish whether retrieval is required or whether frozen sections or IF/ICC are easier (caption: EDTA retrieval; HPA: ICC-IF images). In adipose tissue, distinguish staining from lipid vacuoles and section-edge artefact when scoring adipocytes (HPA: High in adipocytes; standard IHC practice).

HPA tissue IHC evidence for COPS5

Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Supported — 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

TissueCell typeLevelEvidenceSource
Adipose tissue Adipocytes High Protein (IHC) HPA →
Adrenal gland Glandular cells High Protein (IHC) HPA →
Appendix Glandular cells High Protein (IHC) HPA →
Bone marrow Hematopoietic cells High Protein (IHC) HPA →
Breast Glandular cells High Protein (IHC) HPA →

Undetected expression · recommended negative controls

TissueCell typeLevelEvidenceSource
None in HPA: COPS5 is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead.
Section 3

Advanced COPS5 IHC Tips

Use compartment-aware controls and consistent section processing to troubleshoot COPS5 staining in paraffin sections (UniProt Q92905 localisation; standard IHC practice).

Which retrieval conditions should I start with for weak COPS5 staining?
Use heat-mediated antigen retrieval in EDTA buffer at pH 8.0 for paraffin-section COPS5 IHC (datasheet M01849-2). The selected antibody stained a paraffin section after that retrieval, followed by 2 μg/ml primary antibody overnight at 4°C (caption M01849-2). If staining is weak, compare a known staining section processed alongside the test section before changing retrieval conditions (caption M01849-2; standard IHC practice). Keep section thickness, heating, cooling and detection consistent across the comparison so a retrieval change can be judged separately from other workflow differences (standard IHC practice). Avoid treating a stronger, diffuse DAB signal as improved retrieval without checking cellular localisation (HPA tissue IHC; standard IHC practice).
Could fixation explain absent or diffuse COPS5 staining?
Target-specific COPS5 fixation sensitivity is unknown: the selected paraffin-section caption does not state its fixative (caption M01849-2). Record the fixative and fixation duration for each specimen, then compare sections processed with the same retrieval and detection conditions (standard IHC practice; datasheet M01849-2). If a control section stains while a test section does not, review specimen processing and tissue preservation before assigning the difference to COPS5 expression (standard IHC practice). Conversely, a damaged section with diffuse colour cannot establish a fixation-specific effect on this antibody (standard IHC practice). Judge interpretable staining by its cellular distribution and morphology, without inferring fixation tolerance from tissue patterns or protein topology (HPA tissue IHC; UniProt Q92905 topology; standard IHC practice).
Should COPS5 staining be nuclear, cytoplasmic or both?
Assess both nuclear and cytoplasmic DAB staining: tissue IHC reports COPS5 in both compartments across tissues (HPA tissue IHC). UniProt also places COPS5 in the cytosol, nucleus and perinuclear region, with vesicle-associated localisation annotated (UniProt Q92905 localisation). Score nuclear and cytoplasmic staining separately against the counterstained cell morphology, because a single combined score can obscure a compartment shift (HPA tissue IHC; standard IHC practice). Predominantly nuclear staining can be plausible, but a uniformly stained extracellular space is outside these annotated compartments and warrants a background check (UniProt Q92905 localisation; standard IHC practice). IFIT3 can diminish nuclear localisation, so a shift alone does not prove assay failure (UniProt Q92905 localisation).
How should I assess an uncertain antibody epitope in COPS5 IHC?
Do not assign this antibody an epitope from the COPS5 sequence alone; the selected caption gives staining conditions but no epitope position (caption M01849-2). UniProt lists 0 isoforms, an MPN domain at residues 55–192, and no transmembrane segment or annotated glycosylation sites (UniProt Q92905 record). Those annotations help frame specificity questions, but they do not show which residues remain accessible after paraffin processing and retrieval (UniProt Q92905 record; standard IHC practice). Compare staining with an independently characterised antibody where available, and examine whether both reagents identify the same cells and compartments (standard IHC practice; HPA tissue IHC). Interpret discordance as a reason to investigate epitope recognition and staining controls, rather than evidence for an unannotated isoform (UniProt Q92905 isoforms; standard IHC practice).
How can IF help verify a puzzling COPS5 IHC pattern?
Use IF/ICC as a separate cross-check of the cells and compartments identified by chromogenic IHC (standard IF practice; HPA tissue IHC). Multiplex COPS5 with an independently validated marker for the cell type under study; glandular cells are one documented high-staining tissue population for choosing a comparison (HPA tissue IHC). Select spectrally separated fluorophores after checking tissue autofluorescence and include single-colour controls for bleed-through (standard IF practice). Because COPS5 has no transmembrane segment and is annotated in intracellular compartments, permeabilise sufficiently for antibody access to the relevant epitope, whose location is unspecified here (UniProt Q92905 topology and localisation; caption M01849-2). Compare nuclear signal against a nuclear counterstain; HPA reports enhanced nucleoplasmic localisation in its subcellular assay (HPA subcellular; standard IF practice).
What should I check when COPS5 DAB staining is widespread?
Widespread cellular staining can be plausible because HPA reports nuclear and cytoplasmic expression across tissues, but distribution alone does not establish specificity (HPA tissue IHC; standard IHC practice). Inspect a no-primary control for secondary-reagent signal and check the peroxidase block when interpreting DAB colour (standard IHC practice). The selected example used 10% goat serum blocking, a biotinylated secondary reagent and DAB development, so review those steps when troubleshooting that workflow (caption M01849-2). Compare staining within intact cells against the counterstain, especially near folds, edges and damaged areas where nonspecific colour can mislead (standard IHC practice). Titrate primary antibody around the documented 2 μg/ml condition only while holding retrieval and detection constant (caption M01849-2; standard IHC practice).
How should I quantify nuclear and cytoplasmic COPS5 staining? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and tissue region before scoring, and exclude folds, necrosis and poorly preserved areas by a consistent rule (standard IHC practice). Report the percentage of positive cells and, if intensity is reproducible, an H-score for nuclear and cytoplasmic compartments separately (HPA tissue IHC; standard IHC practice). For spatial analyses, report positive-cell density per mm² of evaluable tissue or of the prespecified compartment, with the denominator stated (standard IHC practice). Normalise comparisons to the number of evaluable cells or measured area, and keep retrieval, DAB development and image settings consistent across sections (standard IHC practice; datasheet M01849-2). Record whether compartment shifts accompany intensity changes, since COPS5 is annotated in both nucleus and cytoplasm (UniProt Q92905 localisation).
How do I distinguish credible COPS5 staining from artefact?
Credible staining should resolve to intact cells and be assessed in nuclear and cytoplasmic compartments, which are documented for COPS5 in tissue IHC (HPA tissue IHC; standard IHC practice). Check whether the stained cells match the expected morphology; HPA reports high staining in several populations, including adipocytes and glandular cells, without defining every specimen as positive (HPA tissue IHC). Treat colour concentrated at section edges, folds or necrotic regions as suspect until it persists in well-preserved tissue (standard IHC practice). Use a no-primary control and verify peroxidase blocking when DAB signal appears in implausible locations or obscures the counterstain (standard IHC practice). A nuclear-to-cytoplasmic shift can be biologically plausible in the presence of IFIT3, but IHC localisation alone does not establish that mechanism (UniProt Q92905 localisation; standard IHC practice).
Boster reagents

Best COPS5 / COP9 signalosome complex subunit 5 IHC Antibodies

The catalog shows COPS5 IHC in human tissue and mouse and rat testis, plus IF in T-47D cells (catalog: IHC and IF captions).

Real IHC data IHC analysis of JAB1 using anti-JAB1 antibody (M01849-2). JAB1 was detected in paraffin-embedded section of human esophageal squamous carcinoma tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2μg/ml mouse anti-JAB1 Antibody (M01849-2) overnight at 4°C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1021) with DAB as the chromogen.
Anti-JAB1 Antibody ® (monoclonal, 4G9)
Cat # M01849-2
Real IHC data IHC analysis of JAB1 using anti-JAB1 antibody (PB9490). JAB1 was detected in a paraffin-embedded section of human mammary cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1 μg/ml rabbit anti-JAB1 Antibody (PB9490) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1022) with DAB as the chromogen.
Anti-JAB1/COPS5 Antibody ®
Cat # PB9490
Real IHC data Figure 5. IHC analysis of JAB1/COPS5 using anti-JAB1/COPS5 antibody (M01849). JAB1/COPS5 was detected in a paraffin-embedded section of human breast cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with a dilution of 1:50 rabbit anti-JAB1/COPS5 Antibody (M01849) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.
Anti-JAB1 Monoclonal Antibody
Cat # M01849

M01849-2 has paraffin-section IHC images from human esophageal squamous carcinoma, liver cancer, lung cancer and placenta, plus an IF image from T-47D cells (catalog: M01849-2 image captions). PB9490 has paraffin-section IHC images from human mammary cancer and mouse and rat testis; M01849 has a paraffin-section IHC image from human breast cancer (catalog: PB9490 and M01849 IHC captions).

Which to pick: For tissue IHC across human, mouse and rat samples, choose PB9490: its own captions show paraffin-section staining in all three species, and its listed IHC dilution is 0.5–1 μg/ml (catalog: PB9490 IHC captions and dilution). For IF/ICC, M01849-2 has a fluorescence image in T-47D cells using 5 μg/ml; M01849 lists IF/ICC but has no IF image in this payload (catalog: M01849-2 IF caption; M01849 applications and IF images). M01849 is a rabbit monoclonal IHC option with a human breast cancer paraffin-section image at 1:50; the fixative is unreported in the IHC captions for all three SKUs (catalog: M01849 host, clone and IHC caption; M01849-2, PB9490 and M01849 IHC captions).

Each figure is that product's own IHC / IF validation image from its datasheet.

References

  1. UniProt Consortium. UniProt entry Q92905 (CSN5_HUMAN, COP9 signalosome complex subunit 5).
  2. Human Protein Atlas. COPS5 tissue IHC expression (reliability: Supported).
  3. Human Protein Atlas. COPS5 subcellular location (ICC-IF): Localized to the nucleoplasm..
  4. Human Protein Atlas. COPS5 antibody validation summary (3 antibodies).
  5. COPS5 amplification and overexpression confers tamoxifen-resistance in ERα-positive breast cancer by degradation of NCoR. Nature communications 2016 — PMC4932188.
  6. COP9 signalosome subunit 5 regulates cancer metastasis by deubiquitinating SNAIL. Oncotarget 2018 — PMC5945527.
  7. Increased Jab1/COPS5 is associated with therapeutic response and adverse outcome in lung cancer and breast cancer patients. Oncotarget 2017 — PMC5722579.
  8. Analysis of differential gene expression profile identifies novel biomarkers for breast cancer. Oncotarget 2017 — PMC5777718.
  9. PubMed PMID:8837781 — UniProt-cited evidence.
  10. PubMed PMID:9341143 — UniProt-cited evidence.
  11. PubMed PMID:15489334 — UniProt-cited evidence.