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ALDH3B2 / Aldehyde dehydrogenase family 3 member B2 · IHC design guide
Design Immunohistochemistry for ALDH3B2
Plan ALDH3B2 paraffin-section IHC with the catalog antibody at 1:50–1:200 (datasheet: A05723). Breast adipocytes provide a high-staining reference, but interpret cytoplasmic staining cautiously because antibody staining and RNA expression show low consistency (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
ALDH3B2 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 | Breast adipocytes show high cytoplasmic staining (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen) | |
| Positive control | Breast | |
| Negative control | Adipose tissue+4 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep fixation consistent across paraffin sections. (standard IHC practice; not target-specific) | |
| Caveat | Low consistency between antibody staining and RNA expression (HPA tissue IHC) | |
| Regulation | RNA enhanced in breast, esophagus and skin (HPA) | |
| Isoform / epitope | 0 isoforms; residues 383–385 form a propeptide; epitope unknown (UniProt) |
Section 1
Recommended ALDH3B2 IHC & IF Protocols
The catalog antibody IHC-P protocol (datasheet) is accompanied by published protocols for breast cancer tissue (PMC6776190) and cholangiocarcinoma tissue microarrays (PMC8671409).
| Sample | Paraffin-embedded human liver carcinoma tissue; fixative not specified (datasheet A05723) |
| Fixation | Image fixative and duration unreported (datasheet A05723); 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-ALDH3B2, 1:50-1:200 (datasheet A05723) |
| 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 | ALDH3B2-positive staining in adipocytes of breast (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression in several tissues including breast. No signal in the no-primary control. |
💡Decision noteStart with citrate pH 6.0 at 95–98 °C for 20 min (page retrieval rule); EDTA pH 8.0 is a published alternative (PMC8671409).
Section 2
What Is the Expected ALDH3B2 Staining Pattern?
ALDH3B2 is associated with lipid droplets and has no transmembrane segment (UniProt P48448: localisation and topology). In paraffin section IHC, expect cytoplasmic staining; breast adipocytes are the clearest supplied high staining example (HPA: cytoplasmic tissue profile; High in breast adipocytes). Interpret that expectation cautiously: HPA rates its tissue IHC pattern Approved but reports low consistency with RNA expression and pending external verification (HPA: tissue IHC reliability).
| Breast adipocytes show strong cytoplasmic chromogen, with cell boundaries and counterstained nuclei still readable. | This fits the supplied positive tissue and compartment (HPA: High in breast adipocytes; cytoplasmic profile). Lipid droplet association supports a cytoplasmic interpretation, but routine chromogenic IHC alone cannot prove that individual droplets carry the signal (UniProt P48448: lipid droplet localisation; general IHC practice). |
| Signal is mainly nuclear or follows a sharply outlined cell surface rather than the expected cytoplasmic distribution. | Treat the compartment mismatch as suspicious and compare it with a negative control and the breast positive reference (HPA: cytoplasmic tissue profile; High in breast adipocytes; general IHC practice). UniProt lists no transmembrane segment, so a surface outline alone is weak evidence for the expected target pattern (UniProt P48448: topology). |
| Strong chromogen appears in a cell population recorded as unstained, such as adrenal gland glandular cells. | Consider cross-reactivity or endogenous detection activity before assigning it to ALDH3B2 (HPA: Not detected in adrenal gland glandular cells; general IHC practice). HPA's tissue observations are reference patterns, not proof that every specimen of that cell type must be negative (HPA: tissue IHC profile). |
| Chromogen coats many unrelated structures, obscures cell boundaries, or fills blank areas of the section. | This reads as diffuse background rather than an interpretable cell-specific pattern (general IHC practice). Check the negative control, blocking and wash conditions, and detection system before scoring intensity; broad background can make weakly stained cells look positive (general IHC practice). |
| Breast adipocytes lack a detectable signal while the counterstain and tissue architecture remain visible. | This conflicts with the supplied high staining reference but does not by itself establish true absence of ALDH3B2 (HPA: High in breast adipocytes). Check positive-control performance and antibody and detection steps, then consider specimen variation; HPA flags low staining–RNA consistency and pending external verification (general IHC practice; HPA: tissue IHC reliability). |
💡Expected ALDH3B2 appearanceCall a result supportive when breast adipocytes show clear, high intensity cytoplasmic staining (HPA: High in breast adipocytes; cytoplasmic profile); isolated nuclear staining, a membrane-only rim, or widespread structure-independent chromogen is suspect (UniProt P48448: topology; general IHC practice).
| Cellular compartment | UniProt places ALDH3B2 at lipid droplets and records no transmembrane segment; HPA describes tissue IHC as cytoplasmic (UniProt P48448: localisation and topology; HPA: tissue IHC profile). These observations guide slide interpretation without establishing a droplet-resolved chromogenic pattern (general IHC practice). |
| Tissue context | HPA reports High staining in breast adipocytes but Not detected in adipocytes from adipose tissue (HPA: tissue IHC levels). Keep those tissue contexts distinct when choosing references; the supplied data do not make adipocytes universally positive or negative (HPA: tissue IHC profile). |
| Strength of validation | The supplied antibody HPA045132 has IHC status Approved, while the tissue profile has low consistency with RNA data and awaits external verification (HPA: antibody validation; tissue IHC reliability). Treat an unexpected positive as a finding to check with controls, not as a confirmed expression map (general IHC practice). |
| Processing and epitope limits | UniProt records a 1–382 chain and a 383–385 propeptide, but the supplied record gives no antibody epitope (UniProt P48448: processing; HPA: supplied antibody record). It therefore cannot predict whether processing alters this antibody's IHC signal or justify a target-specific retrieval claim. |
| IF/ICC Q&A: is a membrane pattern established? | HPA's subcellular summary says Membrane but provides no ICC-IF image cell lines or main location (HPA: subcellular record). UniProt places ALDH3B2 at lipid droplets and lists no transmembrane segment (UniProt P48448: localisation and topology). The supplied evidence does not establish a dependable IF/ICC membrane pattern. |
| Situation | Likely cause | Next action |
|---|---|---|
| Breast adipocytes stain weakly or not at all. | A failed IHC run or specimen variation may account for the result; the supplied reference is High, with reliability caveats (HPA: High in breast adipocytes; tissue IHC reliability; general IHC practice). | Review the positive control and the applied IHC antibody, detection and retrieval steps; document any change before comparing staining intensity across runs (general IHC practice). |
| Adipose tissue adipocytes stain strongly. | That disagrees with the supplied Not detected observation for this tissue, despite High staining in breast adipocytes (HPA: adipose tissue and breast IHC levels). | Confirm tissue identity and staining controls, then assess whether the signal is cellular and reproducible; retain the two tissue contexts separately in scoring (general IHC practice; HPA: tissue IHC levels). |
| Signal is predominantly nuclear. | A nuclear pattern does not match the reported cytoplasmic tissue profile or lipid droplet localisation (HPA: tissue IHC profile; UniProt P48448: localisation). | Compare with the breast positive reference and negative control; if the nuclear pattern persists, score it separately as an unexpected pattern rather than expected ALDH3B2 staining (HPA: High in breast adipocytes; general IHC practice). |
| Unrelated cells or blank areas develop chromogen. | Non-specific antibody binding or endogenous detection activity may produce widespread signal (general IHC practice). | Inspect a negative control and the blocking, washing and chromogen steps; interpret cell-specific staining only after background is acceptably low (general IHC practice). |
| A cell-surface rim dominates the stain. | The rim conflicts with HPA's cytoplasmic IHC profile; the UniProt record lists no transmembrane segment (HPA: tissue IHC profile; UniProt P48448: topology). | Check section edges, controls and cell morphology for artefact, and report the rim as an unexpected observation if it remains reproducible (general IHC practice). |
| A result is judged from a single positive field. | HPA's Approved tissue profile has low consistency with RNA expression and awaits external verification (HPA: tissue IHC reliability). | Compare the field with controls and the expected cell and compartment pattern; record tissue context and uncertainty when an observation differs from HPA (HPA: tissue IHC profile; general IHC practice). |
Experiment evidence
IHC & IF images showing ALDH3B2 staining
A05723 · IHCImmunohistochemistry (IHC) analyzes of ALDH3B2 pAb in paraffin-embedded human liver carcinoma tissue at 1:50,showing cytoplasm and membrane
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000132746-ALDH3B2/tissue/cerebral+cortex#imid_11923311
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000132746-ALDH3B2/tissue/colon#imid_11923229
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000132746-ALDH3B2/tissue/liver#imid_11923230
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000132746-ALDH3B2/tissue/kidney#imid_11923239
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for ALDH3B2 IHC & IF
🧪Run breast first and score adipocytes for ALDH3B2 staining (HPA: High in breast adipocytes). Use adipose tissue adipocytes as the negative tissue (HPA: Not detected in adipose tissue adipocytes); on the breast slide, non-adipocyte cells can serve as internal negative references if they show only background signal.
Positive control tissue: Breast (Adipocytes, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA carries no ICC-IF cell line for ALDH3B2; derive a cell-line control from the positive tissue's cell type (Adipocytes) and confirm it by RNA or western blot first.
Technical controls: Include a no-primary, secondary-only control; host- and clonality-matched nonimmune IgG; and an ALDH3B2 knockout biological negative if available. For chromogenic detection, quench endogenous peroxidase and block endogenous biotin when using the captioned avidin–peroxidase detection system (selected A05723 IHC caption: biotinylated secondary followed by avidin–peroxidase).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected A05723 paraffin-section caption does not state a fixative (selected A05723 IHC caption: fixative unreported). Antigen retrieval dependence is unreported, so retrieval conditions need empirical optimization (selected A05723 IHC caption: retrieval unreported). The supplied evidence does not establish whether frozen sections or IF are easier; in breast sections, assess staining specifically in adipocytes and check lipid-rich areas for background artefact (HPA: High in breast adipocytes; UniProt P48448: lipid droplet localization).
HPA tissue IHC evidence for ALDH3B2
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — Low consistency between antibody staining and RNA expression data. Pending external verification.). 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 |
|---|---|---|---|---|
| Breast | Adipocytes | High | 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 → |
| Appendix | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced ALDH3B2 IHC Tips
Troubleshoot ALDH3B2 staining in paraffin sections by checking retrieval, controls, compartment, and scoring before interpreting chromogenic signal.
How should I retrieve ALDH3B2 in paraffin sections when staining is weak?
Start with citrate buffer at pH 6.0 for heat-induced retrieval at 95–98 °C for 20 min (page retrieval rule). Let sections cool consistently, then compare staining with a matched section processed without primary antibody to separate retrieval effects from detection background (standard IHC practice). The selected catalog image used paraffin-embedded human liver carcinoma at 1:50, but its caption does not report retrieval conditions (A05723 caption). If signal remains weak, adjust retrieval time on adjacent sections while keeping antibody dilution and detection constant (standard IHC practice). Record tissue morphology alongside signal, because excessive heating can make a brighter stain harder to interpret (standard IHC practice).
Could fixation explain variable ALDH3B2 staining between paraffin samples?
Target-specific fixation sensitivity is unknown: the selected paraffin-section caption does not state a fixative (A05723 caption). Record the fixative, fixation interval, and processing history for each specimen before comparing staining across blocks (standard IHC practice). Process matched control sections together and hold retrieval, the 1:50 starting dilution, and chromogenic development consistent while evaluating fixation as a possible variable (A05723 caption; standard IHC practice). If staining differs, examine preservation and background as well as signal, then test adjacent sections with a controlled retrieval adjustment (standard IHC practice). Do not assign a fixation effect to ALDH3B2 from its lipid-droplet location or terminal modification alone (UniProt P48448).
Should I score membrane staining as ALDH3B2-positive?
The selected tissue image describes cytoplasmic and membrane staining in paraffin-embedded human liver carcinoma (A05723 caption). UniProt places ALDH3B2 at lipid droplets and reports no transmembrane segment, while HPA describes cytoplasmic tissue staining and lists a membrane subcellular summary (UniProt P48448; HPA tissue IHC; HPA subcellular). Score cytoplasmic and membrane-pattern signal separately rather than treating either pattern as definitive protein location (standard IHC practice). Compare each pattern with a no-primary section and with preserved cellular morphology in the same region (standard IHC practice). A sharply outlined plasma membrane pattern needs independent support before being interpreted as a transmembrane pool (UniProt P48448 topology).
How do processing and epitope uncertainty affect ALDH3B2 IHC interpretation?
The supplied record lists 0 isoforms, a 1–382 protein chain, and a 383–385 propeptide (UniProt P48448). It also notes a modified cysteine at position 382, but the selected antibody caption does not identify its epitope (UniProt P48448; A05723 caption). Therefore, neither loss of a terminal epitope nor selective recognition of a processed form can be assigned to an IHC pattern from these data (UniProt P48448; A05723 caption). If an epitope map becomes available, compare it with the annotated chain boundary before choosing orthogonal confirmation (standard IHC practice). Meanwhile, retain a no-primary control and evaluate staining by compartment and cell type (standard IHC practice).
How can I assess ALDH3B2 by multiplex IF after paraffin-section IHC?
Treat IF as a separate optimization: the supplied HPA subcellular entry lists no cell lines with ICC/IF images, and the selected antibody evidence is a chromogenic paraffin-section image (HPA subcellular; A05723 caption). For breast tissue, pair ALDH3B2 with an adipocyte marker and inspect whether signals occur in the expected cells (HPA tissue IHC; standard IF practice). Choose fluorophores after checking unstained tissue autofluorescence, favoring a cleaner channel for the weaker signal (standard IF practice). Because ALDH3B2 has no transmembrane segment and is assigned to lipid droplets, evaluate permeabilisation for access to its intracellular epitope; the antibody epitope itself is unspecified (UniProt P48448; A05723 caption). Include single-stain and no-primary controls before interpreting overlap (standard IF practice).
What controls help identify nonspecific chromogenic ALDH3B2 staining?
The selected image includes a PBS-instead-of-primary control and uses a biotinylated secondary followed by avidin–peroxidase detection (A05723 caption). Run that no-primary control beside each test section, using the same detection and development conditions (standard IHC practice). Include an endogenous peroxidase block and inspect residual signal before attributing dark deposits to ALDH3B2 (standard chromogenic IHC practice). With a biotin–avidin system, also assess endogenous biotin background where it affects the specimen (A05723 caption; standard IHC practice). If widespread staining persists, shorten chromogen development or titrate the primary around the caption's 1:50 condition, changing one variable at a time (A05723 caption; standard IHC practice).
How should I quantify ALDH3B2 staining across heterogeneous sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the tissue region and cell population before scoring, then exclude folds, edges, and nonviable areas by the same rules for every section (standard IHC practice). For cellular staining, record the percentage of positive cells and intensity categories, or calculate an H-score from those categories (standard IHC practice). If the question concerns focal positive cells, report positive-cell density per mm² of evaluable tissue as a complementary measure (standard IHC practice). Normalise each result to the number of eligible cells or evaluable area, and compare batches using matched controls and development settings (standard IHC practice). Keep cytoplasmic and membrane-pattern scores separate because both appear in the selected image (A05723 caption).
When is an ALDH3B2-positive IHC result biologically credible?
A credible result has a reproducible cellular pattern above the no-primary control and occurs in morphologically intact tissue (standard IHC practice). ALDH3B2 is assigned to lipid droplets without a transmembrane segment, while the selected image reports cytoplasmic and membrane staining; interpret an isolated crisp membrane rim cautiously (UniProt P48448; A05723 caption). Check whether positive cells match the question being tested: HPA reports high staining in breast adipocytes but no detected staining in adipose-tissue adipocytes, with low consistency against RNA data (HPA tissue IHC). Exclude edge staining, necrotic regions, and residual endogenous peroxidase signal before scoring (standard IHC practice). Confirm a surprising compartment or cell population with an independent method (standard IHC practice).
Boster reagents
Best ALDH3B2 / Aldehyde dehydrogenase family 3 member B2 IHC Antibodies
A05723 has a paraffin-section human liver carcinoma IHC figure (catalog image caption); A05723-1 lists IF/ICC and human, mouse, and rat reactivity (catalog applications/reactivity).
Real IHC data
Anti-ALDH3B2 Antibody
Cat # A05723
A05723 is the SKU with a rendered IHC card: its figure shows cytoplasmic and membrane staining in paraffin-embedded human liver carcinoma at 1:50, with PBS replacing primary antibody in the negative control (catalog image caption). The catalog lists human, mouse, and rat reactivity for A05723; the supplied figure documents a human specimen only (catalog reactivity; catalog image caption).
Which to pick: For tissue IHC, choose A05723 when a matching paraffin-section example is useful; its figure uses human liver carcinoma, and the fixative is unreported (catalog image caption). For IF/ICC, choose A05723-1 because those applications are listed at 1:50, though no IF image is supplied (catalog applications/dilution; catalog IF image alts). For cross-species planning, both SKUs are rabbit polyclonals with human, mouse, and rat listed as reactive species; the supplied tissue image supports human staining only (catalog host/reactivity; A05723 image caption).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P48448 (AL3B2_HUMAN, Aldehyde dehydrogenase family 3 member B2).
- Human Protein Atlas. ALDH3B2 tissue IHC expression (reliability: Approved).
- Human Protein Atlas. ALDH3B2 subcellular location (ICC-IF): Membrane.
- Human Protein Atlas. ALDH3B2 antibody validation summary (1 antibodies).
- Identification of new cancer stem cell markers and signaling pathways in HER‑2‑positive breast cancer by transcriptome sequencing. International journal of oncology 2019 — PMC6776190.
- Aldehyde dehydrogenase 3B2 promotes the proliferation and invasion of cholangiocarcinoma by increasing Integrin Beta 1 expression. Cell death & disease 2021 — PMC8671409.
- PubMed PMID:7484374 — UniProt-cited evidence.
- PubMed PMID:8890755 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for ALDH3B2 (Aldehyde dehydrogenase family 3 member B2). Expected localisation Cytoplasmic tissue staining (HPA tissue IHC). Antigen retrieval: Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen). Validated IHC antibody: A05723 at 1:50-1:200 (the product's own IHC validation image is shown). Controls (HPA): positive Breast (Adipocytes); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human liver carcinoma tissue; fixative not specified (datasheet A05723); 2. Fixation: Image fixative and duration unreported (datasheet A05723); 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-ALDH3B2, 1:50-1:200 (datasheet A05723); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep fixation consistent across paraffin sections. (standard IHC practice; not target-specific); Caveat: Low consistency between antibody staining and RNA expression (HPA tissue IHC); Isoform / epitope: 0 isoforms; residues 383–385 form a propeptide; epitope unknown (UniProt); Regulation: RNA enhanced in breast, esophagus and skin (HPA). Evidence: UniProt P48448, HPA tissue IHC · Approved, HPA ICC-IF · Membrane, Datasheet A05723, PMC6776190, PMC8671409. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/ALDH3B2/immunohistochemistry-ihc-guide.