DBI / Acyl-CoA-binding protein · IHC design guide

Design Immunohistochemistry for DBI

Plan chromogenic DBI IHC in paraffin sections using cytoplasmic staining as the tissue pattern (HPA tissue IHC). Compare hepatocytes and kidney tubules with documented low or undetected cell populations, and account for isoform-dependent expression (HPA tissue IHC; UniProt).

Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use

Immunohistochemistry protocol sheet for DBI (IHC for DBI): expected localisation Cytoplasmic in tissue (HPA tissue IHC); ER/Golgi (UniProt), antibody A01267, validated IHC image, and IHC protocol steps — Printable **DBI** IHC protocol sheet — expected localisation Cytoplasmic in tissue (HPA tissue IHC); ER/Golgi (UniProt), antibody A01267, controls and protocol steps. Open the full DBI IHC guide →

Navigate to: Protocols Staining pattern Controls Antibodies

DBI 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 in tissue (HPA tissue IHC); ER/Golgi (UniProt)
	Staining pattern	Cytoplasmic staining in hepatocytes and kidney tubules (HPA tissue IHC)
	Antigen retrieval	Citrate pH 6 HIER, heat-mediated (datasheet A01267)
	Positive control ⓘ	Appendix+4 more · see all
	Negative control ⓘ	Bone marrow+4 more · see all

Important caveats

Reasons your staining may differ from the expected pattern.

	Fixation	Keep fixation consistent across sections. (standard IHC practice; not target-specific)
	Caveat	Placenta image differs from HPA decidual-cell result (datasheet A01267; HPA tissue IHC)
	Regulation	Isoform 2 is high in liver and adipose tissue (UniProt)
	Isoform / epitope	Six isoforms; confirm epitope coverage across variants (UniProt)

Section 1

Recommended DBI IHC & IF Protocols

The catalog antibody’s IHC-P protocol (datasheet A01267) is accompanied by published DBI staining protocols for cartilage and mouse knee joints (PMC12326017) and tumors (PMC11378439).

Recommended immunohistochemistry (IHC-P) protocol parameters
Sample	Paraffin-embedded human placenta tissues; fixative not specified (datasheet A01267)
Fixation	Image fixative and duration unreported (datasheet A01267); verify before use.
Sectioning	4–5 µm sections on charged slides (standard)
Deparaffinisation	Xylene, graded ethanol series to water (standard)
Antigen retrieval	Heat retrieval: Citrate pH 6, 20 min (datasheet A01267)
Peroxidase block	3% H2O2, 10 min, room temperature (standard)
Blocking	10% goat serum (datasheet A01267)
Primary antibody	Rabbit anti-DBI, 0.5-1μg/ml (datasheet A01267)
Primary incubation	Overnight at 4 °C (datasheet A01267)
Detection	Streptavidin-biotin complex (SABC), DAB chromogen (datasheet A01267)
Counterstain	Hematoxylin, blue, dehydrate and mount (standard)
Expected result	DBI-positive staining in glandular cells of appendix (HPA tissue IHC: High). HPA tissue profile: General cytoplasmic expression. No signal in the no-primary control.

💡Decision noteStart with heat-mediated citrate retrieval at pH 6 (datasheet A01267); both published protocols also use citrate at pH 6 (PMC12326017; PMC11378439).

Section 2

What Is the Expected DBI Staining Pattern?

DBI should show predominantly cytoplasmic staining in paraffin sections, including hepatocytes, kidney tubule cells and several glandular cell populations with high HPA staining (HPA: tissue IHC). Its annotated locations are the endoplasmic reticulum and Golgi apparatus, with Golgi localisation dependent on ligand binding; DBI has no transmembrane segment (UniProt P07108). HPA rates the tissue IHC pattern Approved, with medium consistency against RNA expression (HPA: tissue IHC).

What am I looking at on my slide?
Cytoplasmic stain is strong in hepatocytes or kidney tubule cells, while nuclei remain relatively clear.	This fits the reported high staining in those cells and the general cytoplasmic profile (HPA: tissue IHC). Compare cell identity and compartment before scoring intensity; a brown deposit anywhere in the section is not, by itself, a DBI-positive cell (general IHC practice).
Staining is concentrated in nuclei, or appears only as a sharp cell-surface outline.	Neither pattern matches HPA's general cytoplasmic profile or UniProt's endoplasmic reticulum and Golgi annotations (HPA: tissue IHC; UniProt P07108). Treat it as a localisation mismatch and check controls and detection conditions before assigning it to DBI; the sources do not provide a nuclear or membrane IHC pattern.
Strong stain appears in bone-marrow hematopoietic cells or smooth muscle cells.	Those cell populations were reported as not detected in the HPA tissue IHC record (HPA: tissue IHC). Unexpected stain could reflect nonspecific antibody binding or endogenous detection activity (general IHC practice). It is a reason to investigate, rather than proof that every stained cell is a false positive.
Brown colour covers stroma, lumina and multiple cell types without clear cell boundaries.	A widespread deposit is difficult to reconcile with the reported cell-associated cytoplasmic pattern (HPA: tissue IHC). Review the no-primary control and background across the section; inadequate blocking, residual detection reagents or chromogen development can obscure a specific signal (general IHC practice).
A liver section has no convincing hepatocyte signal.	HPA reports high staining in hepatocytes, so an entirely blank result is unexpected under a working assay (HPA: tissue IHC). Verify that the section contains intact hepatocytes, then review antibody, retrieval and detection controls (general IHC practice). A blank slide alone does not establish absent DBI expression.

💡Expected DBI appearanceCall a positive result when identifiable hepatocytes, kidney tubule cells or listed glandular cells show clear cytoplasmic staining, potentially high in the HPA-listed populations; isolated nuclear, surface-only or cell-free deposit should prompt a background check (HPA: tissue IHC; UniProt P07108; general IHC practice).

How each factor affects the staining
Compartment and topology	HPA describes general cytoplasmic staining; UniProt annotates endoplasmic reticulum and Golgi localisation and no transmembrane segment (HPA: tissue IHC; UniProt P07108). A surface-only IHC pattern therefore needs independent scrutiny. UniProt notes that Golgi localisation depends on ligand binding, so a discrete Golgi accent is not required in every cell (UniProt P07108).
Tissue and cell context	HPA reports high staining in hepatocytes, kidney tubule cells, type II alveolar cells and specified glandular and glial cells, but not detected staining in several other listed populations (HPA: tissue IHC). These are useful comparisons within identified cell types; HPA also reports low tissue RNA specificity, so an unlisted tissue is not automatically negative (HPA: tissue IHC).
Isoforms and epitope coverage	UniProt lists 6 DBI isoforms, with isoform-specific expression descriptions, but the supplied record gives no antibody epitope or isoform coverage (UniProt P07108). Differences among tissues cannot be assigned to isoform recognition from these data. The annotated protein chain spans residues 2–87; this does not establish which sequence the antibody recognises (UniProt P07108).
Validation and IF/ICC Q&A	Q: Does this IHC pattern establish an IF/ICC localisation? A: No. HPA lists antibody HPA051428 as IHC Approved, with no ICC status, no ICC-IF images and no available main subcellular location in its ICC-IF record (HPA: antibodies; HPA: subcellular). Use the IHC pattern as context, without treating it as IF/ICC validation.
Fixation and retrieval evidence	The supplied HPA and UniProt records do not report DBI-specific fixation sensitivity or an antigen-retrieval setting (HPA: tissue IHC; UniProt P07108). Retrieval can be optimised as a general paraffin-section IHC step, but these sources do not support a claim that fixation or a particular retrieval method changes DBI staining (general IHC practice).

Why is my staining missing, weak or wrong?
Situation	Likely cause	Next action
Known-positive hepatocytes are blank.	An assay or section issue is possible because hepatocytes are HPA high; the blank result does not identify which step failed (HPA: tissue IHC).	Confirm hepatocyte preservation and run the assay controls. Review the antibody application, retrieval, detection reagents and chromogen steps without assuming a DBI-specific retrieval requirement (general IHC practice).
All tissue compartments stain brown, including cell-free areas.	The pattern exceeds HPA's general cytoplasmic description and may reflect detection background (HPA: tissue IHC; general IHC practice).	Inspect the no-primary control, blocking and reagent rinses; reassess chromogen development before scoring any cell as positive (general IHC practice).
Signal is nuclear or restricted to cell outlines.	The compartment conflicts with HPA's cytoplasmic profile and UniProt's endoplasmic reticulum and Golgi locations (HPA: tissue IHC; UniProt P07108).	Check morphology and counterstain, compare a known-positive tissue and review background controls. Record the mismatch rather than scoring it as the expected DBI pattern (general IHC practice).
Bone-marrow hematopoietic cells stain as strongly as hepatocytes.	HPA reports hematopoietic cells as not detected and hepatocytes as high; nonspecific binding or endogenous detection activity is possible (HPA: tissue IHC; general IHC practice).	Compare matched controls and cell identity, then reassess blocking and the detection system. Do not use the contrast alone to declare biological expression or a validated negative result (general IHC practice).
A listed positive tissue shows only weak staining.	HPA levels describe observed staining, not a guaranteed intensity in every section; its IHC reliability is Approved with medium RNA concordance (HPA: tissue IHC).	Score the correct cell population and cytoplasmic compartment first. Compare assay controls and section quality before changing interpretation or optimising routine IHC conditions (general IHC practice).
An IF/ICC image is requested to confirm the paraffin-section result.	The supplied HPA record has no ICC status, ICC-IF images or main ICC-IF location for DBI (HPA: antibodies; HPA: subcellular).	Report the IHC result with its tissue and compartment evidence. Treat any IF/ICC result as a separate validation question; the supplied sources provide no IF/ICC protocol or image benchmark (HPA: tissue IHC; HPA: subcellular).

Source: UniProt P07108 Human Protein Atlas tissue Human Protein Atlas subcellular

Experiment evidence

IHC & IF images showing DBI staining

IHC analysis of DBI using anti-DBI antibody (A01267). DBI was detected in paraffin-embedded section of human placenta tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-DBI Antibody (A01267) 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.

A01267 · IHCIHC analysis of DBI using anti-DBI antibody (A01267). DBI was detected in paraffin-embedded section of human placenta tissues. Heat mediated

IHC analysis of DBI using anti-DBI antibody (A01267). DBI was detected in paraffin-embedded section of human prostatic cancer tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-DBI Antibody (A01267) 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.

A01267 · IHCIHC analysis of DBI using anti-DBI antibody (A01267). DBI was detected in paraffin-embedded section of human prostatic cancer tissues. Heat

IF analysis of DBI using anti-DBI antibody (A01267). DBI was detected in immunocytochemical section of U20S cell. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 2μg/mL rabbit anti-DBI Antibody (A01267) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG (BA1127) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

A01267 · IFIF analysis of DBI using anti-DBI antibody (A01267). DBI was detected in immunocytochemical section of U20S cell. Enzyme antigen retrieval w

HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000155368-DBI/tissue/cerebral+cortex#imid_16494147

HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000155368-DBI/tissue/colon#imid_16494081

HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000155368-DBI/tissue/liver#imid_16494074

HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000155368-DBI/tissue/kidney#imid_16494083

← swipe / scroll for more images →

Real IHC / IF data behind the staining-pattern expectations in this section.

Sample controls for DBI IHC & IF

🧪Run appendix first and score glandular cells for DBI staining (HPA: High in appendix glandular cells). Use oral mucosa squamous epithelial cells as the negative tissue (HPA: Not detected in oral mucosa squamous epithelial cells); on the appendix slide, require any cells designated as internal negatives to remain at background, because the supplied HPA row establishes expression only for glandular cells (HPA: appendix glandular cells High).

Positive control tissue: Appendix (Glandular cells, HPA High)

Negative control tissue: Bone marrow (HPA Not detected)

ICC-IF cell lines (HPA subcellular resource): HPA carries no ICC-IF cell line for DBI; derive a cell-line control from the positive tissue's cell type (Glandular cells) and confirm it by RNA or western blot first.

Technical controls: Include a no-primary, secondary-only control, a concentration-matched rabbit IgG isotype control matched to the catalog antibody’s clonality (catalog caption: rabbit anti-DBI antibody), and DBI-knockout material or an immunizing-peptide block if available. Quench endogenous peroxidase and check for endogenous biotin signal in appendix sections when using the caption’s biotin–streptavidin/DAB detection system (catalog caption: biotinylated secondary, SABC and DAB).

⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the exact A01267 paraffin-section caption does not state a fixative (catalog caption: fixative not stated). The caption uses citrate retrieval at pH 6 for 20 minutes, but does not establish that retrieval is required (catalog caption: citrate pH 6, 20 minutes). No supplied evidence shows that frozen sections or IF are easier; avoid using placenta as an uncomplicated negative because the caption shows DBI staining in placenta tissue while HPA reports decidual cells as not detected (catalog caption: placenta staining; HPA: placenta decidual cells Not detected).

HPA tissue IHC evidence for DBI

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
Appendix	Glandular cells	High	Protein (IHC)	HPA →
Breast	Glandular cells	High	Protein (IHC)	HPA →
Caudate	Glial cells	High	Protein (IHC)	HPA →
Epididymis	Glandular cells	High	Protein (IHC)	HPA →
Gallbladder	Glandular cells	High	Protein (IHC)	HPA →

Undetected expression · recommended negative controls

Tissue	Cell type	Level	Evidence	Source
Bone marrow	Hematopoietic cells	Not detected	Protein (IHC)	HPA →
Fallopian tube	Glandular cells	Not detected	Protein (IHC)	HPA →
Oral mucosa	Squamous epithelial cells	Not detected	Protein (IHC)	HPA →
Placenta	Decidual cells	Not detected	Protein (IHC)	HPA →
Smooth muscle	Smooth muscle cells	Not detected	Protein (IHC)	HPA →

Section 3

Advanced DBI IHC Tips

These DBI troubleshooting answers use the selected paraffin section IHC example, the protein record, and tissue staining data to guide interpretation.

What retrieval should I start with for DBI staining in paraffin sections?

Start with heat mediated retrieval in citrate buffer at pH 6 for 20 minutes (datasheet A01267). That condition accompanies DBI staining in a paraffin embedded human placenta section, although the caption does not identify the fixative (datasheet A01267). If staining is weak, first check that heating, cooling, and section handling are consistent across the run (standard IHC practice). Compare any retrieval adjustment with a section processed under the stated citrate condition, using the same antibody concentration and detection steps (standard IHC practice). Score staining by cell and compartment, because DBI has a general cytoplasmic tissue profile (HPA tissue IHC).

How should I troubleshoot DBI staining after fixation?

The selected paraffin section caption does not state a fixative, so DBI specific sensitivity to fixation is unknown (datasheet A01267). Record the fixative, fixation duration, tissue thickness, and processing history for every specimen before comparing staining (standard IHC practice). Use sections processed together when assessing whether a fixation change improves signal, and keep retrieval at citrate pH 6 for 20 minutes as the reference condition (datasheet A01267; standard IHC practice). Review morphology alongside DAB staining, since damaged or poorly preserved areas can confound chromogenic interpretation (standard IHC practice). Do not attribute a fixation effect to DBI without a controlled comparison.

Where should convincing DBI staining appear in tissue?

Evaluate DBI primarily as cellular cytoplasmic staining, the pattern reported across tissues (HPA tissue IHC). Endoplasmic reticulum and Golgi localisation are recorded for DBI, with Golgi localisation dependent on ligand binding (UniProt P07108). Because DBI has no transmembrane segment, a sharply outlined plasma membrane pattern warrants review of staining specificity and tissue morphology (UniProt P07108 topology; standard IHC practice). Compare signal within identified cells against nearby background rather than treating every brown deposit as cytoplasmic expression (standard IHC practice). HPA reports high staining in kidney tubule cells and liver hepatocytes, which can serve as cell specific reference patterns (HPA tissue IHC).

Can this IHC result identify a DBI isoform or modified epitope?

A DBI stain alone cannot assign an isoform unless the antibody epitope and its coverage of the variants are established (standard IHC interpretation). The record lists 6 isoforms and an acyl CoA binding domain spanning residues 2–87 (UniProt P07108). It also lists modifications including acetylation and succinylation, but supplies no evidence that these alter this antibody’s tissue staining (UniProt P07108; datasheet A01267). Isoforms 1–3 have differing reported tissue expression patterns, so a change in intensity need not identify a particular variant (UniProt P07108). Consult epitope documentation and use an independently validated isoform specific assay if that distinction matters (standard IHC practice).

How can I investigate DBI by multiplex immunofluorescence?

Treat IF as a separate validation exercise, because the selected antibody evidence describes chromogenic staining of a paraffin section (datasheet A01267). Pair DBI with a validated marker for the cell population under study; HPA reports high DBI staining in kidney tubule cells and liver hepatocytes (HPA tissue IHC; standard IF practice). Choose fluorophores and filters after checking tissue autofluorescence in unstained sections, and include single stain controls to assess bleed through (standard IF practice). DBI lacks a transmembrane segment and has endoplasmic reticulum and Golgi localisation, so assess permeabilisation for access to intracellular epitopes (UniProt P07108; standard IF practice). Check cell identity and intracellular pattern before interpreting colocalisation (standard IF practice).

How do I distinguish DBI signal from chromogenic background?

Inspect a section without primary antibody alongside the DBI stained section to identify detection reagent and tissue background (standard IHC practice). The selected example used 10% goat serum block, 1 μg/ml primary antibody overnight at 4°C, and biotin based DAB detection (datasheet A01267). If diffuse brown staining persists, check the peroxidase block, reagent controls, washes, and detection exposure before changing the primary concentration (standard IHC practice). Compare suspicious deposits with intact cellular cytoplasm, because general cytoplasmic expression is the reported DBI tissue pattern (HPA tissue IHC). Record whether signal follows tissue edges or damaged regions rather than cell boundaries (standard IHC practice).

What should I measure when comparing DBI IHC across samples? ⚠ ANSWER MARKED FOR VERIFICATION

Define the cell population and tissue region before scoring, then apply the same threshold and imaging settings to every section (standard IHC practice). For cellular staining, report the percentage of positive target cells and an H-score that combines staining intensity with that percentage (standard IHC practice). If counting positive cells, express counts as density per mm² of viable target tissue and report the area measured (standard IHC practice). Normalise comparisons to the number of evaluable target cells or the viable target tissue area, rather than the whole section when composition varies (standard IHC practice). Record cytoplasmic staining separately from background (HPA tissue IHC; standard IHC practice).

What makes a DBI positive stain credible?

A credible result has reproducible cellular cytoplasmic staining with appropriate morphology and a low signal in the no primary control (HPA tissue IHC; standard IHC practice). HPA reports high DBI staining in kidney tubule cells and liver hepatocytes, while placental decidual cells are reported as not detected (HPA tissue IHC). The selected antibody caption shows staining in a human placenta section but does not assign it to decidual cells, so identify the stained cell population before comparing those observations (datasheet A01267; HPA tissue IHC). Question isolated membrane outlines, section edge deposits, necrotic regions, and signal present in enzyme or detection controls (UniProt P07108 topology; standard IHC practice).

Boster reagents

Best DBI / Acyl-CoA-binding protein IHC Antibodies

A01267 has IHC images from human placenta and prostatic cancer paraffin sections, plus an IF/ICC image from U20S cells (catalog image captions).

Real IHC data

Anti-Diazepam Binding Inhibitor/DBI Antibody ®

Cat # A01267

View product →

A01267 was shown on human placenta and prostatic cancer paraffin sections using IHC (A01267 IHC image captions). A01267 was also shown in U20S cells using IF/ICC and is listed as human-reactive (A01267 IF image caption; catalog applications and reactivity).

Which to pick: Choose A01267 for paraffin-section IHC: its captions show staining in human placenta and prostatic cancer at 1 μg/ml after citrate retrieval; the fixative is unreported (A01267 IHC image captions). The same SKU is listed for IF/ICC at 2 μg/ml and has a U20S cell IF image (catalog applications and dilution; A01267 IF image caption). For other species, the payload provides no validated choice because A01267 lists human reactivity only (catalog reactivity).

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

References

UniProt Consortium. UniProt entry P07108 (ACBP_HUMAN, Acyl-CoA-binding protein).
Human Protein Atlas. DBI tissue IHC expression (reliability: Approved).
Human Protein Atlas. DBI subcellular location (ICC-IF): Highest expression in ASC52telo: 1281.3 nTPM.
Human Protein Atlas. DBI antibody validation summary (1 antibodies).
Neutralization of acyl CoA binding protein (ACBP) for the experimental treatment of osteoarthritis. Cell death and differentiation 2025 — PMC12326017.
Sex differences in central endozepine expression and regulation of appetite. Physiology & behavior 2025 — PMC12381991.
Kinetics of apoptosis and expression of apoptosis-related proteins in rat CA3 hippocampus cells after experimental diffuse brain injury. Cell biochemistry and biophysics 2013 — PMC3838593.
Acyl-coenzyme a binding protein (ACBP) - a risk factor for cancer diagnosis and an inhibitor of immunosurveillance. Molecular cancer 2024 — PMC11378439.
PubMed PMID:3020548 — UniProt-cited evidence.
PubMed PMID:2881742 — UniProt-cited evidence.
PubMed PMID:16055366 — UniProt-cited evidence.