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DCAF1 / DDB1- and CUL4-associated factor 1 · IHC design guide
Design Immunohistochemistry for DCAF1
Plan DCAF1 paraffin IHC with catalog antibody A31776 at 2–5 μg/mL (datasheet A31776). Use lung alveolar type II cells as a high-staining reference and score cytoplasmic signal (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
DCAF1 Immunohistochemistry Experimental Design Guide
Expected localisation, validated protocols, controls and antibodies — the at-a-glance facts below, then the full design guide.
Must know before staining
| Expected localisation | Cytoplasmic tissue staining (HPA tissue IHC) | |
| Staining pattern | Many tissue cell types show cytoplasmic staining (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A31776) | |
| Positive control | Lung+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 fixation consistent across sections (standard IHC practice; not target-specific) | |
| Caveat | Staining and RNA have medium consistency (HPA tissue IHC) | |
| Regulation | Testis-enhanced RNA (HPA tissue RNA) | |
| Isoform / epitope | 3 isoforms; epitope coverage is unspecified (UniProt) |
Section 1
Recommended DCAF1 IHC & IF Protocols
The catalog antibody uses heat-mediated EDTA retrieval at pH 8.0 (datasheet A31776). The published IHC examples below describe FFPE tissue staining (PMC7407567; PMC10526264).
| Sample | Paraffin-embedded human placenta tissue; fixative not specified (datasheet A31776) |
| Fixation | Image fixative and duration unreported (datasheet A31776); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat retrieval: EDTA pH 8.0 (datasheet A31776); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A31776) |
| Primary antibody | Rabbit anti-DCAF1, 2-5μg/ml (datasheet A31776) |
| Primary incubation | Overnight at 4 °C (datasheet A31776) |
| Detection | Streptavidin-biotin complex (SABC), DAB chromogen (datasheet A31776) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | DCAF1-positive staining in alveolar cells type II of lung (HPA tissue IHC: High). HPA tissue profile: Ubiquitous cytoplasmic expression. No signal in the no-primary control. |
💡Decision noteStart with EDTA pH 8.0 for the catalog antibody (datasheet A31776); evaluate Tris-EDTA pH 9 when adapting the published VPRBP protocol (PMC7407567).
Section 2
What Is the Expected DCAF1 Staining Pattern?
DCAF1 is an intracellular protein without a transmembrane segment (UniProt Q9Y4B6 topology). In paraffin tissue IHC, expect broadly distributed cytoplasmic staining, especially strong in lung alveolar type II cells (HPA: ubiquitous cytoplasmic expression; High in alveolar type II cells). Interpret this pattern with care: HPA rates tissue IHC Approved, with medium consistency between staining and RNA expression (HPA: tissue IHC reliability).
| Clear cytoplasmic signal in alveolar type II cells, with weaker signal in several other tissue cell types. | This fits the strongest listed tissue result and the broad cytoplasmic IHC profile (HPA: High in lung alveolar type II cells; Medium in several other cell types). Compare intensity within a slide using morphology and counterstain; an HPA level is a reference observation, not a universal scoring threshold (general IHC interpretation). |
| Predominantly nuclear staining in tissue, with little cytoplasmic signal. | An exclusively nuclear tissue pattern conflicts with the reported cytoplasmic IHC profile (HPA: ubiquitous cytoplasmic expression). Check the antibody and detection controls before calling it specific. Nuclear DCAF1 remains biologically plausible (UniProt Q9Y4B6: nucleus); HPA ICC-IF supports nucleoplasmic localization, so compartment alone cannot prove an artefact (HPA: nucleoplasm supported). |
| Strong staining appears in a cell population scored Low by HPA, or in structures that do not resemble intact cells. | Recheck cell identity and slide morphology before assigning DCAF1 expression (general IHC interpretation). For example, HPA lists prostate glandular cells and smooth muscle cells as Low (HPA: tissue IHC). Unexpected strong signal may reflect cross-reactivity or endogenous detection activity (general IHC practice); the HPA listing cannot diagnose its cause. |
| A diffuse wash of chromogen obscures cell borders and both positive and low-staining areas. | This cannot be read as the reported cell-associated cytoplasmic pattern (HPA: ubiquitous cytoplasmic expression). Diffuse background can arise from nonspecific binding, incomplete washing, or detection background (general IHC practice). Judge DCAF1 only where individual cells and compartments remain distinguishable (general IHC interpretation). |
| No staining is visible in lung alveolar type II cells. | That misses a listed High reference cell type (HPA: High in lung alveolar type II cells). First confirm those cells are present and identifiable, then evaluate slide-wide technical failure with appropriate controls (general IHC practice). A negative result in one preparation alone does not establish absence of DCAF1, given HPA's medium staining-to-RNA consistency (HPA: tissue IHC reliability). |
💡Expected DCAF1 appearanceCall a tissue IHC result positive when intact alveolar type II cells show clear, strong cytoplasmic chromogen, with variable weaker staining elsewhere (HPA: High in alveolar type II cells; ubiquitous cytoplasmic expression); diffuse cell-independent color is suspect background (general IHC interpretation).
| Tissue and cell choice | Lung alveolar type II cells offer a High staining reference; adipocytes, adrenal and breast glandular cells, and bronchial respiratory epithelial cells are listed Medium (HPA: tissue IHC). Prostate glandular cells, ovarian stromal cells, smooth muscle cells, and bone marrow hematopoietic cells are listed Low (HPA: tissue IHC). Low means a weak reference, not a proven DCAF1-null control (HPA: tissue IHC). |
| Compartment and assay | Tissue IHC is described as ubiquitously cytoplasmic (HPA: tissue IHC), while DCAF1 is annotated in cytoplasm, nucleus, and centrosome (UniProt Q9Y4B6: subcellular location). These observations set different expectations for different preparations; tissue chromogen alone cannot resolve fine nuclear structures or establish centrosomal localization (general microscopy interpretation). |
| Antibody validation | HPA036142 and HPA053203 are Approved for IHC; HPA052445 has no listed IHC status (HPA: antibody validation). Approved tissue reliability carries a medium staining-to-RNA consistency description (HPA: tissue IHC reliability). Use validation status to choose evidence, while checking the actual slide against morphology and controls (general IHC practice). |
| Isoforms and epitope scope | UniProt lists three DCAF1 isoforms and a full-length chain spanning residues 1–1507 (UniProt Q9Y4B6: isoforms; processing). No antibody epitope or isoform coverage is supplied here; an IHC pattern cannot be assigned to a particular isoform from these records alone (UniProt Q9Y4B6: isoforms; HPA: antibody listings). |
| IF/ICC expectation? | For IF/ICC, HPA reports mainly nucleoplasmic signal with an additional nucleoli fibrillar-center location, both supported (HPA: subcellular ICC-IF). That is the IF/ICC interpretation here; it does not replace the cytoplasmic tissue IHC reference (HPA: tissue IHC). |
| Situation | Likely cause | Next action |
|---|---|---|
| Lung reference cells are unstained while the whole slide is pale. | A slide-wide failure is possible, but the observation alone does not identify which step failed (general IHC practice; HPA: High in alveolar type II cells). | Verify alveolar type II cells by morphology and counterstain, then inspect primary-antibody, detection, and run controls; repeat the run if controls failed (general IHC practice). Do not infer DCAF1 absence from a failed reference. |
| A strong nuclear-only tissue signal replaces the expected cytoplasmic pattern. | The result is discordant with HPA tissue IHC, although nuclear localization is recorded by UniProt and supported in HPA ICC-IF (HPA: cytoplasmic tissue IHC; UniProt Q9Y4B6: nucleus; HPA: nucleoplasm supported). | Compare compartments in intact cells, review the antibody's IHC validation, and check detection controls (general IHC practice; HPA: antibody validation). Report the discordance rather than treating nuclear signal alone as proof of specificity. |
| Low-reference cell types stain as strongly as the lung reference. | The contrast differs from HPA's listed High and Low observations; nonspecific binding or endogenous detection activity is possible (HPA: tissue IHC levels; general IHC practice). | Confirm cell identity and compare matched sections with an appropriate negative detection control (general IHC practice). Interpret any residual staining cautiously because HPA levels are observations, not absolute exclusions (HPA: tissue IHC). |
| Diffuse color persists outside recognizable cells. | Nonspecific reagent binding, insufficient washing, or endogenous detection activity may contribute (general IHC practice). This pattern cannot establish the reported cytoplasmic localization (HPA: tissue IHC). | Inspect a control that omits the primary antibody, check washing and blocking, and assess the detection chemistry's endogenous-activity control as appropriate (general IHC practice). Reassess only cell-associated signal. |
| Only scattered weak cytoplasmic cells are visible in a broadly positive tissue. | Cell composition and low staining intensity may limit interpretation; HPA's broad profile does not promise equal intensity in every cell (HPA: ubiquitous cytoplasmic expression; HPA: tissue IHC levels). | Identify the stained cell population on the counterstained section, compare it with the relevant HPA cell-level reference, and verify run controls before scoring the tissue (general IHC practice; HPA: tissue IHC). |
| IF/ICC looks nucleoplasmic while the tissue IHC slide looks cytoplasmic. | The two reported assay observations differ: nucleoplasm is supported in ICC-IF, whereas the tissue IHC profile is cytoplasmic (HPA: subcellular ICC-IF; HPA: tissue IHC). | Document each assay with its own compartment reference and inspect its controls (general microscopy practice). Do not score a tissue IHC slide against the ICC-IF location alone (HPA: tissue IHC; HPA: subcellular ICC-IF). |
Experiment evidence
IHC & IF images showing DCAF1 staining
A31776 · IHCIHC analysis of VPRBP/DCAF1 using anti-VPRBP/DCAF1 antibody (A31776). VPRBP/DCAF1 was detected in paraffin-embedded section of human placent
A31776 · IHCIHC analysis of VPRBP/DCAF1 using anti-VPRBP/DCAF1 antibody (A31776). VPRBP/DCAF1 was detected in paraffin-embedded section of human breast
A31776 · IFIF analysis of VPRBP/DCAF1 using anti-VPRBP/DCAF1 antibody (A31776). VPRBP/DCAF1 was detected in immunocytochemical section of PC-3 cells. E
A31776-1 · IHCIHC analysis of VPRBP/DCAF1 using anti-VPRBP/DCAF1 antibody (A31776-1). VPRBP/DCAF1 was detected in a paraffin-embedded section of human chr
A31776-1 · IHCIHC analysis of VPRBP/DCAF1 using anti-VPRBP/DCAF1 antibody (A31776-1). VPRBP/DCAF1 was detected in a paraffin-embedded section of human col
A31776-1 · IFIF analysis of VPRBP/DCAF1 using anti-VPRBP/DCAF1 antibody (A31776-1). VPRBP/DCAF1 was detected in an immunocytochemical section of Hela cel
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000145041-DCAF1/tissue/cerebral+cortex#imid_19065207
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000145041-DCAF1/tissue/colon#imid_19065125
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000145041-DCAF1/tissue/liver#imid_19065118
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000145041-DCAF1/tissue/kidney#imid_19065127
HPA · ICC-IF HeLaHuman Protein Atlas immunofluorescence, HeLa, antibody HPA052445; green = DCAF1, blue = nucleus, red = microtubules.
HPA · ICC-IF HeLaHuman Protein Atlas immunofluorescence, HeLa, antibody HPA052445; green = DCAF1, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for DCAF1 IHC & IF
🧪Start with lung and assess staining in alveolar type II cells (HPA: High in alveolar cells type II); cells without specific chromogen should appear counterstain-only, but should not be treated as proven DCAF1-negative cells (UniProt Q9Y4B6: ubiquitously expressed). No negative tissue is available because DCAF1 was detected in all 45 scored tissues, so use no-primary and isotype controls to establish negative staining (HPA: no negative rows).
Positive control tissue: Lung (Alveolar cells type II, HPA High)
Negative control tissue: None in HPA: DCAF1 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 DCAF1 in HeLa, PC-3, U2OS, CACO-2, NIH 3T3, with annotated localisation: Nucleoplasm (supported) (HPA subcellular).
Technical controls: Run a no-primary, secondary-only control and a host-matched rabbit IgG isotype control, and confirm specificity with DCAF1 knockout material if available (A31776 caption: rabbit primary antibody; standard IHC practice). For chromogenic lung IHC, block endogenous peroxidase and inspect the controls for tissue pigment or background signal (standard IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected paraffin-section caption does not state a fixative (A31776 caption). The caption uses heat retrieval in EDTA at pH 8.0, which provides a starting condition but does not establish that retrieval is required (A31776 caption). The supplied evidence does not establish whether frozen sections or IF are easier; lung pigment and endogenous peroxidase warrant control-slide review for apparent chromogenic signal (standard IHC practice).
HPA tissue IHC evidence for DCAF1
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — 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 |
|---|---|---|---|---|
| Lung | Alveolar cells type II | High | Protein (IHC) | HPA → |
| Adipose tissue | Adipocytes | Medium | Protein (IHC) | HPA → |
| Adrenal gland | Glandular cells | Medium | Protein (IHC) | HPA → |
| Appendix | Glandular cells | Medium | Protein (IHC) | HPA → |
| Breast | Glandular cells | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| None in HPA: DCAF1 is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead. | ||||
Section 3
Advanced DCAF1 IHC Tips
Troubleshoot DCAF1 staining in paraffin sections by checking retrieval, controls, cellular localisation and scoring before interpreting changes in chromogenic signal.
Which retrieval condition should I try first for weak DCAF1 staining?
Start with heat mediated retrieval in EDTA at pH 8.0 for paraffin sections (datasheet A31776). The documented placenta IHC workflow used that condition before incubation with 2 µg/mL primary antibody overnight at 4°C (datasheet A31776). If staining is weak, vary heating duration within the instrument’s recommended range, then compare matched sections processed together; excessive retrieval can damage tissue morphology or increase background (standard IHC practice). Keep antibody concentration, detection chemistry and imaging conditions fixed during that comparison so a change in signal can be attributed more confidently to retrieval (standard IHC practice).
Could fixation explain inconsistent DCAF1 staining between paraffin blocks?
The selected tissue IHC caption identifies a paraffin section but does not state its fixative, so target specific fixation sensitivity is unknown (datasheet A31776). Record each block’s fixative and fixation duration, then compare sections using the same pH 8.0 EDTA retrieval and detection run (datasheet A31776; standard IHC practice). Variable fixation can alter epitope accessibility and tissue morphology in paraffin IHC generally; this is a workflow concern, not a demonstrated DCAF1 specific effect (standard IHC practice). Include a consistently processed reference section in each run before assigning differences between blocks to biology (standard IHC practice).
Should DCAF1 appear nuclear, cytoplasmic or both in tissue sections?
Assess nuclear and cytoplasmic staining separately: DCAF1 is annotated in both compartments, with cell cycle dependent chromatin association described by similarity (UniProt Q9Y4B6 localisation). Tissue IHC reports ubiquitous cytoplasmic expression, whereas cell imaging supports nucleoplasmic localisation and an additional nucleolar fibrillar center signal (HPA tissue IHC; HPA subcellular). These sources use different specimens and assays, so score the compartment visible in each cell population rather than treating one pattern as universally required (standard IHC interpretation). Inspect hematoxylin counterstain and section morphology before calling a faint nuclear deposit specific, and document mixed patterns explicitly (standard IHC practice).
How do I evaluate an unexpected DCAF1 staining pattern without a mapped epitope?
DCAF1 has 3 annotated isoforms, but the supplied catalog IHC caption does not identify the antibody epitope (UniProt Q9Y4B6 isoforms; datasheet A31776). Its chromo domain spans residues 562–593 and its LisH domain spans 846–878; these positions alone cannot establish which isoforms the antibody detects (UniProt Q9Y4B6 domains and isoforms). Check the antibody’s epitope documentation before interpreting differences as isoform specific, and compare serial sections with an independently validated reagent if available (standard IHC practice). Because DCAF1 has no annotated transmembrane segment, a crisp membrane only pattern warrants closer evaluation of background and tissue boundaries (UniProt Q9Y4B6 topology; standard IHC interpretation).
How should I follow up DCAF1 IHC with multiplex immunofluorescence?
Treat IF as a separate assay: the selected antibody evidence describes chromogenic IHC in a paraffin section, while HPA cell imaging supports mainly nucleoplasmic DCAF1 localisation (datasheet A31776; HPA subcellular). Pair DCAF1 with a validated marker for the cell type being evaluated, and check each channel alone before interpreting overlap (standard IF practice). Choose fluorophores and filter sets that keep the DCAF1 channel distinct from tissue autofluorescence, using unstained and single stain controls (standard IF practice). DCAF1 lacks a transmembrane segment, so use a controlled permeabilisation step when assessing intracellular epitopes, adjusting it to preserve nuclear morphology (UniProt Q9Y4B6 topology; standard IF practice).
What should I check when DCAF1 DAB staining looks diffuse?
Compare the section with a no primary control to identify signal from the detection system, and inspect the hematoxylin counterstain for tissue damage (standard IHC practice). The documented workflow used 10% goat serum, biotinylated goat anti rabbit secondary antibody and a streptavidin biotin complex with DAB (datasheet A31776). Check endogenous peroxidase blocking and, with this biotin based system, endogenous biotin controls before calling widespread brown deposition DCAF1 staining (standard IHC practice). If background persists, titrate primary antibody around the documented 2 µg/mL condition while holding retrieval, wash conditions and DAB development time constant (datasheet A31776; standard IHC practice).
How should I quantify DCAF1 staining across tissue samples? ⚠ ANSWER MARKED FOR VERIFICATION
Define the tissue region and cell population before scoring, then report the percentage of positive cells and an intensity weighted H score when cellular staining can be resolved (standard IHC practice). Score nuclear and cytoplasmic compartments separately because both are annotated for DCAF1, and record the threshold used to call a cell positive (UniProt Q9Y4B6 localisation; standard IHC practice). For spatial comparisons, report positive cells per mm² and normalise to evaluable tissue area or the number of eligible cells, excluding folds and necrosis (standard IHC practice). Process comparison sections together and apply the same illumination, segmentation and scoring rules throughout (standard IHC practice).
How can I distinguish genuine DCAF1 staining from artefact?
Look for reproducible cellular staining in intact regions: DCAF1 is annotated in cytoplasm and nucleus, and tissue IHC reports ubiquitous cytoplasmic expression (UniProt Q9Y4B6 localisation; HPA tissue IHC). A membrane only rim is unexpected for a protein without a transmembrane segment, so examine tissue boundaries and detection controls before interpreting it biologically (UniProt Q9Y4B6 topology; standard IHC interpretation). Compare cell identity and morphology with the counterstain; edge deposits, necrotic regions and endogenous enzyme activity can mimic chromogenic positivity (standard IHC practice). Use matched controls and independently processed sections to confirm a pattern, particularly because HPA rates agreement between staining and RNA expression as medium (HPA tissue IHC; standard IHC practice).
Boster reagents
Best DCAF1 / DDB1- and CUL4-associated factor 1 IHC Antibodies
Two catalog antibodies have IHC images from human paraffin sections; one also has mouse and rat brain IHC images (catalog IHC captions). Both have human cell IF/ICC images (catalog IF captions).
Real IHC data
Anti-VPRBP/DCAF1 Antibody ®
Cat # A31776
Real IHC data
Anti-VPRBP/DCAF1 Antibody ®
Cat # A31776-1
A31776 has paraffin-section IHC images from human placenta, breast cancer, ovarian adenocarcinoma and pancreatic cancer (A31776 IHC captions), plus an IF/ICC image from PC-3 cells (A31776 IF caption). A31776-1 has paraffin-section IHC images from human chronic tonsillitis, human colonic adenocarcinoma, mouse brain and rat brain (A31776-1 IHC captions), plus an IF/ICC image from HeLa cells (A31776-1 IF caption).
Which to pick: For human tissue IHC, choose the SKU with an image closest to your specimen; both use EDTA retrieval at pH 8.0 and 2 μg/ml primary antibody in their own paraffin-section captions (A31776 and A31776-1 IHC captions). For IF/ICC, both have human cell images at 5 μg/ml; choose A31776 for the PC-3 example or A31776-1 for the HeLa example (A31776 and A31776-1 IF captions). For mouse or rat tissue IHC, A31776-1 has direct brain-section images, while A31776 lists mouse and rat reactivity without those species in its IHC captions; clonality and the paraffin sections’ fixative are unreported (catalog reactivity, clone fields and IHC captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q9Y4B6 (DCAF1_HUMAN, DDB1- and CUL4-associated factor 1).
- Human Protein Atlas. DCAF1 tissue IHC expression (reliability: Approved).
- Human Protein Atlas. DCAF1 subcellular location (ICC-IF): Mainly localized to the nucleoplasm. In addition localized to the nucleoli fibrillar center..
- Human Protein Atlas. DCAF1 antibody validation summary (3 antibodies).
- CRL4-DCAF1 ubiquitin E3 ligase directs protein phosphatase 2A degradation to control oocyte meiotic maturation. Nature communications 2015 — PMC4557334.
- The Potential of MLN3651 in Combination with Selumetinib as a Treatment for Merlin-Deficient Meningioma. Cancers 2020 — PMC7407567.
- DCAF1 interacts with PARD3 to promote hepatocellular carcinoma progression and metastasis by activating the Akt signaling pathway. Journal of experimental & clinical cancer research : CR 2024 — PMC11071249.
- VprBP/DCAF1 Triggers Melanomagenic Gene Silencing through Histone H2A Phosphorylation. Biomedicines 2023 — PMC10526264.
- PubMed PMID:9872452 — UniProt-cited evidence.
- PubMed PMID:12168954 — UniProt-cited evidence.
- PubMed PMID:16641997 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for DCAF1 (DDB1- and CUL4-associated factor 1). Expected localisation Cytoplasmic tissue staining (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A31776). Validated IHC antibody: A31776 at 2-5μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Lung (Alveolar cells type II); negative None in HPA (detected in all 45 tissues); use no-primary + isotype controls. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human placenta tissue; fixative not specified (datasheet A31776); 2. Fixation: Image fixative and duration unreported (datasheet A31776); verify before use.; 3. Sectioning: 4–5 µm sections on charged slides (standard); 4. Deparaffinisation: Xylene, graded ethanol series to water (standard); 5. Antigen retrieval: Heat retrieval: EDTA pH 8.0 (datasheet A31776); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A31776); 8. Primary antibody: Rabbit anti-DCAF1, 2-5μg/ml (datasheet A31776); 9. Primary incubation: Overnight at 4 °C (datasheet A31776). Watch for: Fixation: Keep fixation consistent across sections (standard IHC practice; not target-specific); Caveat: Staining and RNA have medium consistency (HPA tissue IHC); Isoform / epitope: 3 isoforms; epitope coverage is unspecified (UniProt); Regulation: Testis-enhanced RNA (HPA tissue RNA). Evidence: UniProt Q9Y4B6, HPA tissue IHC · Approved, HPA ICC-IF · Mainly localized to the nucleoplasm. In addition localized to the…, Datasheet A31776, PMC7407567, PMC10526264. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/DCAF1/immunohistochemistry-ihc-guide.