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DNAJB1 / DnaJ homolog subfamily B member 1 · IHC design guide
Design Immunohistochemistry for DNAJB1
Use nuclear and cytoplasmic staining as the DNAJB1 tissue IHC benchmark (HPA tissue IHC). Adrenal glandular cells offer a high-staining reference (HPA tissue IHC); score nuclear and cytoplasmic signal separately because heat shock can shift DNAJB1 toward nucleoli (UniProt).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
DNAJB1 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 in tissue (HPA tissue IHC) | |
| Staining pattern | General nuclear and cytoplasmic staining (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A03100-1) | |
| Positive control | Adrenal gland+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 tissue fixation consistent across samples (standard IHC practice; not target-specific) | |
| Caveat | Heat shock can shift DNAJB1 toward nucleoli (UniProt) | |
| Regulation | Heat shock shifts nuclear signal (UniProt) | |
| Isoform / epitope | 2 isoforms; epitope coverage unspecified (UniProt) |
Section 1
Recommended DNAJB1 IHC & IF Protocols
The catalog antibody’s IHC-P protocol (datasheet A03100-1) appears alongside published DNAJB1 IHC protocols for fish ovarian frozen sections (PMC6485077) and liver sections (PMC11278829).
| Sample | Paraffin-embedded human gastric signet ring cell carcinoma tissue; fixative not specified (datasheet A03100-1) |
| Fixation | Image fixative and duration unreported (datasheet A03100-1); 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 A03100-1); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A03100-1) |
| Primary antibody | Rabbit anti-DNAJB1, 2-5 μg/ml (datasheet A03100-1) |
| Primary incubation | Overnight at 4 °C (datasheet A03100-1) |
| Detection | Streptavidin-biotin complex (SABC), DAB chromogen (datasheet A03100-1) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | DNAJB1-positive staining in glandular cells of adrenal gland (HPA tissue IHC: High). HPA tissue profile: General nuclear and cytoplasmic expression. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA retrieval at pH 8.0 for the catalog antibody (datasheet A03100-1). The fish frozen-section protocol used citrate at pH 6.0 (PMC6485077).
Section 2
What Is the Expected DNAJB1 Staining Pattern?
DNAJB1 is a soluble protein found in the cytoplasm and nucleus; it has no transmembrane segment (UniProt P25685 topology and subcellular location). Expect nuclear and cytoplasmic staining across several cell types, with high staining in selected epithelial, glandular and follicle cells (HPA tissue IHC). The tissue IHC profile has Enhanced reliability, reflecting high consistency between antibody staining and RNA expression (HPA tissue IHC).
| Nuclear and cytoplasmic staining in bronchial respiratory epithelial cells or ovarian follicle cells. | This fits the general tissue pattern and the High level reported for those cells (HPA tissue IHC). Judge intensity against matched controls; a positive stain alone does not establish a particular cellular stress state (general IHC interpretation). |
| Predominantly membrane outlining or extracellular deposits, with little cellular staining. | That distribution conflicts with the reported cytoplasmic and nuclear locations and lack of a transmembrane segment (UniProt P25685; HPA tissue IHC). Review morphology, detection controls and antibody specificity before assigning it to DNAJB1 (general IHC practice). |
| Strong staining in cardiomyocytes or smooth muscle cells while expected positive cells stain weakly. | Those cell types are listed as Low, so this contrast warrants review; Low does not mean absent (HPA tissue IHC). Check cell identification and controls for cross-reactivity or endogenous chromogenic activity (general IHC practice). |
| Diffuse color over tissue, empty spaces and the slide, without clear cell boundaries. | Noncellular background cannot be read as the nuclear and cytoplasmic pattern (HPA tissue IHC). Compare a no-primary control and assess blocking, washes and detection chemistry (general IHC practice). |
| No staining in a selected High reference cell population, such as adrenal glandular cells. | This fails the expected reference pattern (HPA tissue IHC). Check section quality, retrieval and reagent performance with a known-positive control before interpreting other tissues as negative (general IHC practice). |
💡Expected DNAJB1 appearanceCall a result positive when epithelial, glandular or follicle cells show interpretable nuclear and cytoplasmic staining, potentially High in the listed reference tissues (HPA tissue IHC); membrane-only outlines or noncellular color are suspect because DNAJB1 is reported in the cytoplasm and nucleus and lacks a transmembrane segment (UniProt P25685; HPA tissue IHC).
| Compartment and stress state | DNAJB1 can move rapidly from cytoplasm to nucleus, especially nucleoli, after heat shock (UniProt P25685). Record handling and compare like conditions before treating a nuclear shift as an assay failure (general IHC interpretation). |
| Tissue and cell selection | HPA reports Low tissue RNA specificity and a general nuclear and cytoplasmic IHC profile; its named High and Low cell populations provide useful references, not universal positive and negative tissues (HPA tissue IHC). |
| Antibody validation | HPA063247 is IHC Enhanced and CAB017450 is IHC Supported (HPA antibodies). These are antibody-specific validation labels; apply the label only to the antibody actually used (HPA antibodies). |
| Isoforms and antigen design | Two isoforms are listed, but the supplied record does not map an IHC epitope to either one (UniProt P25685). Do not infer isoform-specific staining from this record; consult the selected antibody's documented immunogen before making that claim. |
| IF/ICC comparison | What should IF/ICC show? Mainly nucleoplasm, with additional cytosol, in the HPA cell images (HPA subcellular ICC-IF). Treat this as a localisation cross-check; the HPA tissue IHC levels describe sections, not IF/ICC intensity. |
| Situation | Likely cause | Next action |
|---|---|---|
| Known High reference cells are blank. | The assay may have failed, or the selected cells may be missing from the section (general IHC practice). | Verify the cell population and run a known-positive section; review the selected antibody's documented retrieval and detection conditions (general IHC practice). High reference examples include bronchial respiratory epithelial cells (HPA tissue IHC). |
| Only a crisp membrane rim appears. | The pattern disagrees with DNAJB1's nuclear and cytoplasmic locations and absent transmembrane segment (UniProt P25685). | Compare a no-primary control, inspect morphology and confirm the antibody's IHC validation before scoring the rim as target staining (general IHC practice; HPA antibodies). |
| Strong color appears in a Low reference population. | Cross-reactivity or endogenous detection activity is possible, but Low is not equivalent to negative (HPA tissue IHC; general IHC practice). | Confirm cell identity; compare no-primary and appropriate detection controls alongside a High reference population before assigning specificity (general IHC practice; HPA tissue IHC). |
| Haze obscures nuclei and cytoplasm. | Background from blocking, washes or detection may prevent compartment scoring (general IHC practice). | Compare a no-primary control, review blocking and washes, and adjust detection conditions according to the selected assay's instructions (general IHC practice). |
| Staining is much more nuclear in one sample. | Stress-dependent nuclear and nucleolar translocation is possible; the stain alone cannot establish its cause (UniProt P25685). | Compare sample handling and matched controls, then score nuclear and cytoplasmic compartments separately (general IHC practice). |
| IF/ICC localisation seems different from tissue IHC. | The HPA ICC-IF summary emphasizes nucleoplasm with additional cytosol, whereas tissue IHC reports general nuclear and cytoplasmic expression (HPA subcellular ICC-IF; HPA tissue IHC). | Compare compartments within each application and check the antibody's application-specific validation; do not transfer tissue IHC intensity categories to IF/ICC (HPA antibodies; HPA tissue IHC). |
Experiment evidence
IHC & IF images showing DNAJB1 staining
A03100-1 · IHCIHC analysis of Hsp40/DNAJB1 using anti-Hsp40/DNAJB1 antibody (A03100-1). Hsp40/DNAJB1 was detected in a paraffin-embedded section of human
A03100-1 · IFIF analysis of Hsp40/DNAJB1 using anti-Hsp40/DNAJB1 antibody (A03100-1). Hsp40/DNAJB1 was detected in an immunocytochemical section of A549
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000132002-DNAJB1/tissue/cerebral+cortex#imid_17680383
HPA · EsophagusHuman Protein Atlas IHC, Esophagus. Source: https://www.proteinatlas.org/ENSG00000132002-DNAJB1/tissue/esophagus#imid_17680339
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000132002-DNAJB1/tissue/colon#imid_17680309
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000132002-DNAJB1/tissue/liver#imid_17680310
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody CAB017450; green = DNAJB1, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody CAB017450; green = DNAJB1, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for DNAJB1 IHC & IF
🧪Run adrenal gland first and score its glandular cells for staining (HPA: High in adrenal gland glandular cells). HPA lists no negative tissue and detects DNAJB1 in all 45 scored tissues, so no-primary and isotype controls carry the negative comparison; no resident cell type on the positive slide is established as DNAJB1-negative, and cells lacking specific signal should show counterstain without DAB deposit (HPA: no negative tissue; standard IHC practice).
Positive control tissue: Adrenal gland (Glandular cells, HPA High)
Negative control tissue: None in HPA: DNAJB1 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 DNAJB1 in A-431, U-251MG, U2OS, A-549, PC-3, with annotated localisation: Nucleoplasm (supported) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control and a matched nonimmune rabbit IgG isotype control for the rabbit catalog antibody; use a validated DNAJB1 knockout specimen or peptide block, if available, as a biological specificity control (caption: rabbit primary antibody; standard IHC practice). For the caption’s SABC/DAB workflow, quench endogenous peroxidase and assess endogenous biotin to distinguish tissue background from specific staining (caption: SABC/DAB; standard IHC practice).
⚠️Feasibility: No target-specific fixation window or fixation effect is reported, and the selected A03100-1 paraffin-section caption does not state the fixative (caption: fixative unreported). The caption documents heat-mediated retrieval in EDTA at pH 8.0, but does not establish that this retrieval is required for every specimen (caption: EDTA retrieval). The supplied evidence does not establish that frozen sections or IF are easier, and reports no adrenal-specific artefact; IF/ICC images do support nucleoplasmic and cytosolic localization as a separate readout (HPA: subcellular localization and ICC-IF images).
HPA tissue IHC evidence for DNAJB1
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — High 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 |
|---|---|---|---|---|
| Adrenal gland | Glandular cells | High | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | High | Protein (IHC) | HPA → |
| Cervix | Squamous epithelial cells | High | Protein (IHC) | HPA → |
| Esophagus | Squamous epithelial cells | High | Protein (IHC) | HPA → |
| Gallbladder | Glandular cells | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| None in HPA: DNAJB1 is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead. | ||||
Section 3
Advanced DNAJB1 IHC Tips
Troubleshoot DNAJB1 staining in paraffin sections by checking retrieval, compartment, controls, and scoring against its documented expression pattern (UniProt P25685; HPA tissue IHC).
How should I retrieve DNAJB1 in paraffin sections when staining is weak?
Start with heat-mediated retrieval in EDTA at pH 8.0 for paraffin sections (datasheet A03100-1). The selected tissue image used this retrieval before overnight incubation with 2 μg/ml primary antibody at 4°C; its caption does not specify retrieval time or temperature (caption A03100-1). If staining remains weak, vary heating and cooling times on matched sections while holding antibody concentration and detection constant, and assess tissue integrity alongside signal (standard IHC practice). Compare nuclear and cytoplasmic staining in the same cell population, because both compartments are documented for DNAJB1 (HPA tissue IHC; UniProt P25685).
Could fixation explain weak or uneven DNAJB1 staining?
Target-specific fixation sensitivity is unknown: the selected DNAJB1 paraffin-section caption does not state a fixative (caption A03100-1). Record each sample's fixative, fixation duration, section thickness, and processing history before attributing differences in nuclear or cytoplasmic signal to biology (standard IHC practice; HPA tissue IHC). For a controlled comparison, process matched specimens consistently, apply EDTA retrieval at pH 8.0, and stain them in the same run (datasheet A03100-1; standard IHC practice). If morphology or staining varies across a section, inspect processing and section quality, then compare intact interior regions rather than inferring a DNAJB1-specific fixation effect (standard IHC practice).
Should DNAJB1 staining be nuclear, cytoplasmic, or nucleolar?
Score nuclear and cytoplasmic staining separately in chromogenic sections, because general expression in both compartments is reported for DNAJB1 (HPA tissue IHC). Nucleoplasmic localisation with additional cytosolic localisation is supported by cell imaging, while UniProt also lists the nucleolus (HPA subcellular; UniProt P25685). UniProt reports rapid movement from cytoplasm to nucleus, especially nucleoli, after heat shock; document stress conditions before treating a changed compartment ratio as a biological result (UniProt P25685). Review high-power fields with a nuclear counterstain and compare matched controls, since DAB overdevelopment can obscure whether brown signal lies in the nucleus or surrounding cytoplasm (standard IHC practice).
How do isoforms and epitope location affect DNAJB1 IHC interpretation?
DNAJB1 has 2 annotated isoforms, a J domain at residues 2–70, and a reported phosphothreonine at residue 307 (UniProt P25685). The supplied antibody caption does not identify its epitope, so a positive chromogenic stain cannot distinguish isoforms or establish phosphorylation at that residue (caption A03100-1; UniProt P25685). Check the antibody's epitope documentation before making either claim, and compare any stated sequence with both isoforms and the modified site (standard antibody validation practice; UniProt P25685). Until that mapping is available, report staining as DNAJB1 immunoreactivity, with compartment and cell type recorded separately (HPA tissue IHC; standard IHC practice).
How can IF help verify the cell type and compartment seen by IHC?
Use IF as an orthogonal check on the IHC pattern by multiplexing DNAJB1 with a validated marker for the cell population being scored and a nuclear counterstain (standard IF practice). Choose a fluorophore in a channel with low tissue autofluorescence, and include single-label and secondary-only controls to assess bleed-through and background (standard IF practice). DNAJB1 has no transmembrane segment and is reported in nucleoplasm and cytosol, so use a validated permeabilisation step to expose intracellular epitopes without assuming an extracellular-facing target (UniProt P25685 topology; HPA subcellular; standard IF practice). Compare cell-by-cell compartment patterns with chromogenic sections, while recognising that the supplied paraffin IHC caption does not validate an IF protocol (caption A03100-1).
How can I separate DNAJB1 signal from chromogenic background?
Run a no-primary control through the same secondary, detection, and DAB steps to identify staining independent of the DNAJB1 antibody (standard IHC practice). The selected paraffin-section workflow used 10% goat serum, 2 μg/ml primary overnight at 4°C, and a biotinylated secondary with SABC and DAB (caption A03100-1). Include an appropriate peroxidase block and assess whether the biotin-based detection system contributes background in the specimen; these are general chromogenic workflow checks (standard IHC practice). If controls are clean but staining is diffuse, titrate primary concentration and DAB development while preserving the documented EDTA pH 8.0 retrieval as the starting condition (datasheet A03100-1; standard IHC practice).
What should I measure when comparing DNAJB1 IHC across samples? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring, because DNAJB1 can stain nuclei and cytoplasm (HPA tissue IHC). For each compartment, record the percentage of positive cells and staining intensity, then calculate an H-score if intensity categories are reproducible across matched slides (standard IHC practice). Normalise positive-cell counts to the number of evaluable cells in the same population, or report positive cells per mm² of viable tissue when cell counts are impractical (standard IHC practice). Keep retrieval, imaging, threshold, and DAB development consistent, and exclude necrotic or damaged areas from the denominator (datasheet A03100-1; standard IHC practice).
What makes a DNAJB1-positive IHC result biologically credible?
Look for reproducible nuclear or cytoplasmic staining in intact cells, since both compartments are documented for DNAJB1 (HPA tissue IHC; UniProt P25685). A high-staining reference may include adrenal glandular cells or bronchial respiratory epithelial cells, while hippocampal neuronal cells and heart cardiomyocytes have reported low staining; assess the actual sampled cell type (HPA tissue IHC). Treat edge-restricted signal, necrotic regions, and staining reproduced in a no-primary control as possible processing or detection artefacts (standard IHC practice). Check for endogenous peroxidase contribution and compare matched sections before interpreting unusual compartment shifts as a stress response, despite reported heat-shock-associated nuclear and nucleolar translocation (standard IHC practice; UniProt P25685).
Boster reagents
Best DNAJB1 / DnaJ homolog subfamily B member 1 IHC Antibodies
A03100-1 has IHC data from human gastric signet ring cell carcinoma paraffin sections and IF data from A549 cells; listed reactivity covers human, mouse, and rat (catalog image captions and reactivity).
Real IHC data
Anti-Hsp40/DNAJB1 Antibody ®
Cat # A03100-1
The card for A03100-1 will show chromogenic IHC in a human gastric signet ring cell carcinoma paraffin section (A03100-1 IHC image caption). Its IF image shows staining in A549 cells, and the catalog lists both IF and ICC applications (A03100-1 IF image caption; catalog applications).
Which to pick: Choose A03100-1 for tissue IHC: its own image shows a paraffin section with EDTA pH 8.0 retrieval, 2 μg/ml primary antibody, and DAB detection; the fixative is unreported (A03100-1 IHC image caption). For IF/ICC, choose A03100-1 based on its A549 cell IF image and listed IF and ICC applications (A03100-1 IF image caption; catalog applications). For cross-species work, A03100-1 lists human, mouse, and rat reactivity, though its supplied IHC image and IHC dilution listing cover human samples; its host is rabbit and clonality is unreported (catalog reactivity, host, and dilution; A03100-1 IHC image caption).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P25685 (DNJB1_HUMAN, DnaJ homolog subfamily B member 1).
- Human Protein Atlas. DNAJB1 tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. DNAJB1 subcellular location (ICC-IF): Mainly localized to the nucleoplasm. In addition localized to the cytosol..
- Human Protein Atlas. DNAJB1 antibody validation summary (2 antibodies).
- Molecular cloning and characteristics of DnaJa1and DnaJb1 in Coilia nasus: possible function involved in oogenesis during spawning migration. BMC developmental biology 2019 — PMC6485077.
- A Prognostic Prediction Model Developed Based on Four CpG Sites and Weighted Correlation Network Analysis Identified DNAJB1 as a Novel Biomarker for Pancreatic Cancer. Frontiers in oncology 2020 — PMC7477361.
- CRISPR/Cas9 Engineering of Adult Mouse Liver Demonstrates That the Dnajb1-Prkaca Gene Fusion Is Sufficient to Induce Tumors Resembling Fibrolamellar Hepatocellular Carcinoma. Gastroenterology 2017 — PMC5801691.
- Diagnosis and Molecular Characterization of Potential RNA Binding Protein Involved in the Pathogenesis of Liver Ischemia Reperfusion Injury. Journal of inflammation research 2024 — PMC11278829.
- PubMed PMID:1754405 — UniProt-cited evidence.
- PubMed PMID:8250930 — UniProt-cited evidence.
- PubMed PMID:8975727 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for DNAJB1 (DnaJ homolog subfamily B member 1). Expected localisation Nuclear and cytoplasmic in tissue (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A03100-1). Validated IHC antibody: A03100-1 at 2-5 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Adrenal gland (Glandular cells); 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 gastric signet ring cell carcinoma tissue; fixative not specified (datasheet A03100-1); 2. Fixation: Image fixative and duration unreported (datasheet A03100-1); verify before use.; 3. Sectioning: 4–5 µm sections on charged slides (standard); 4. Deparaffinisation: Xylene, graded ethanol series to water (standard); 5. Antigen retrieval: Heat retrieval: EDTA pH 8.0 (datasheet A03100-1); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A03100-1); 8. Primary antibody: Rabbit anti-DNAJB1, 2-5 μg/ml (datasheet A03100-1); 9. Primary incubation: Overnight at 4 °C (datasheet A03100-1). Watch for: Fixation: Keep tissue fixation consistent across samples (standard IHC practice; not target-specific); Caveat: Heat shock can shift DNAJB1 toward nucleoli (UniProt); Isoform / epitope: 2 isoforms; epitope coverage unspecified (UniProt); Regulation: Heat shock shifts nuclear signal (UniProt). Evidence: UniProt P25685, HPA tissue IHC · Enhanced, HPA ICC-IF · Mainly localized to the nucleoplasm. In addition localized to the…, Datasheet A03100-1, PMC6485077, PMC11278829. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/DNAJB1/immunohistochemistry-ihc-guide.