This website uses cookies to ensure you get the best experience on our website.
- Table of Contents
AHSA1 / Activator of 90 kDa heat shock protein ATPase homolog 1 · IHC design guide
Design Immunohistochemistry for AHSA1
Plan chromogenic AHSA1 IHC in paraffin sections using fallopian tube glandular cells as a high-staining reference (HPA tissue IHC). Start the catalog antibody at 2–5 μg/ml and compare with salivary gland glandular cells, where staining was not detected (datasheet A05733-2; HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
AHSA1 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 staining in most tissues (HPA tissue IHC) | |
| Staining pattern | Cytoplasmic signal across most tissue cell types (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A05733-2) | |
| Positive control | Fallopian tube+4 more · see all | |
| Negative control | Salivary gland+2 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 | Staining has medium consistency with RNA data (HPA tissue IHC) | |
| Regulation | Expression regulation unreported (UniProt) | |
| Isoform / epitope | 2 isoforms; epitope differences unreported (UniProt) |
Section 1
Recommended AHSA1 IHC & IF Protocols
The catalog antibody’s IHC-P protocol (datasheet A05733-2) and a published protocol for oral tissues (PMC12519801) provide starting points for AHSA1 staining.
| Sample | Paraffin-embedded human breast infiltrating ductal carcinoma tissue; fixative not specified (datasheet A05733-2) |
| Fixation | Image fixative and duration unreported (datasheet A05733-2); 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 A05733-2); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A05733-2) |
| Primary antibody | Rabbit anti-AHSA1, 2-5 μg/ml (datasheet A05733-2) |
| Primary incubation | Overnight at 4 °C (datasheet A05733-2) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A05733-2) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | AHSA1-positive staining in glandular cells of fallopian tube (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression in most tissues. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA pH 8.0 retrieval (datasheet A05733-2); try citrate retrieval when optimizing staining (PMC12519801).
Section 2
What Is the Expected AHSA1 Staining Pattern?
AHSA1 should stain mainly in the cytoplasm, consistent with its cytosolic localization and lack of a transmembrane segment (UniProt O95433; HPA: cytoplasmic expression in most tissues). Expect prominent staining in fallopian tube glandular cells, tonsil germinal center cells, and late spermatids (HPA: High). HPA rates its tissue IHC evidence Supported, with medium consistency between antibody staining and RNA expression (HPA).
| Clear cytoplasmic staining in fallopian tube glandular cells, tonsil germinal center cells, or late spermatids. | This matches the strongest listed tissue IHC patterns (HPA: High in each cell population). Assess the named cells within the section; neighboring cell types need not have the same intensity (HPA: cell-specific tissue IHC entries). |
| Predominantly nuclear staining, or a sharp plasma membrane outline, with little cytoplasmic signal. | Question specificity: AHSA1 is principally cytosolic (HPA: cytosol; UniProt O95433: cytosol). UniProt reports no transmembrane segment and only a possible transient ER interaction, which does not establish a dominant membrane pattern (UniProt O95433). |
| Strong staining in salivary glandular cells, skeletal myocytes, or smooth muscle cells while expected positive cells are weak. | These cells were not detected in the listed HPA tissue IHC observations (HPA: Not detected). Check cross-reactivity and chromogenic detection background before calling the signal AHSA1 (standard IHC practice); a single negative reference is not proof of biological absence. |
| Uniform color across cells, stroma, and empty spaces, with poor cell boundaries. | Treat diffuse staining as background until controls show a cell-associated pattern (standard IHC practice). AHSA1's reported tissue pattern is cytoplasmic and varies by cell type (HPA); uniform slide-wide color is not evidence of broader expression. |
| No staining in the named cells of a known-positive tissue. | A blank result conflicts with the listed high-staining references (HPA: fallopian tube glandular cells, tonsil germinal center cells, late spermatids). First verify that the expected cells are present and that the detection run worked (standard IHC practice); do not infer AHSA1 loss from one section. |
💡Expected AHSA1 appearanceCall a positive result when distinct cytoplasmic chromogen is strong in the named high-staining cells (HPA: High; HPA: cytoplasmic); predominantly nuclear, membrane-outline, or uniform cell-free color warrants a specificity or background check (HPA: cytosol; standard IHC practice).
| Cell population and tissue | HPA reports High staining in three named populations, Medium in several others, and Not detected in selected glandular and muscle cells (HPA: tissue IHC). Score the relevant cells rather than assigning one intensity to an entire section (standard IHC practice). |
| Compartment | Cytosol is supported by ICC-IF, and cytoplasmic expression is reported across most tissues (HPA: subcellular; HPA: tissue IHC). UniProt allows transient ER interaction, so a minor reticular component alone cannot establish a separate, stable localization (UniProt O95433). |
| Antibody evidence | Both listed antibodies have Supported IHC status (HPA: HPA000903 and CAB006244). Overall tissue IHC reliability is Supported with medium staining–RNA consistency (HPA); interpret unexpected staining against tissue controls rather than treating validation status as proof for every section. |
| Isoforms and modifications | Two isoforms and several modified residues are annotated (UniProt O95433). The payload gives no epitope mapping or evidence that either feature changes IHC staining; do not assign an unusual pattern to an isoform or modification without separate validation. |
| What should ICC-IF show? | A cytosolic pattern is the supported reference, with images listed for A-431, U-251MG, and U2OS (HPA: subcellular ICC-IF). ICC-IF is a localization cross-check; this section provides no IF/ICC protocol option. |
| Situation | Likely cause | Next action |
|---|---|---|
| Positive tissue appears blank. | The expected cell population may be absent from the section, or the IHC run may have failed (standard IHC practice). | Confirm the relevant cells are present; check run controls and the catalog antibody's IHC-P instructions before interpreting absence (standard IHC practice; HPA: named High populations). |
| Nuclei dominate the signal. | The compartment disagrees with supported cytosolic localization (HPA: cytosol; UniProt O95433: cytosol). | Compare with a known-positive section and reagent controls; score AHSA1 only where cell-associated cytoplasmic staining is credible (standard IHC practice; HPA: cytoplasmic tissue expression). |
| Muscle or salivary glandular cells stain strongly. | This conflicts with the listed Not detected observations and may reflect cross-reactivity or detection background (HPA: tissue IHC; standard IHC practice). | Check a no-primary control and compare the result with a high-staining reference population before accepting the unexpected signal (standard IHC practice; HPA: High populations). |
| Chromogen covers the section diffusely. | Nonspecific antibody binding or endogenous detection activity can obscure a cellular pattern (standard chromogenic IHC practice). | Review blocking, washes, detection controls, and counterstain; seek defined cytoplasmic staining in the named cells (standard IHC practice; HPA: cytoplasmic tissue expression). |
| Only faint staining appears in liver cholangiocytes or lung macrophages. | These are listed as Low, so weak signal there alone does not indicate assay failure (HPA: tissue IHC). | Judge run sensitivity using a named High population in an appropriate control section, while scoring the low-level cells separately (HPA: tissue IHC; standard IHC practice). |
| Two validated antibodies give different intensities. | Supported IHC status does not establish identical staining strength, and HPA reports only medium staining–RNA consistency overall (HPA: antibody validation; HPA: reliability). | Compare compartment and cell distribution against the same tissue controls; document disagreement without assigning an epitope or fixation cause that the supplied evidence does not establish. |
Experiment evidence
IHC & IF images showing AHSA1 staining
A05733-2 · IHCIHC analysis of AHA1/AHSA1 using anti-AHA1/AHSA1 antibody (A05733-2). AHA1/AHSA1 was detected in a paraffin-embedded section of human breast
A05733-2 · IHCIHC analysis of AHA1/AHSA1 using anti-AHA1/AHSA1 antibody (A05733-2). AHA1/AHSA1 was detected in a paraffin-embedded section of human colore
A05733-2 · IFIF analysis of AHA1/AHSA1 using anti-AHA1/AHSA1 antibody (A05733-2). AHA1/AHSA1 was detected in an immunocytochemical section of A431 cells.
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000100591-AHSA1/tissue/cerebral+cortex#imid_549632
HPA · HippocampusHuman Protein Atlas IHC, Hippocampus. Source: https://www.proteinatlas.org/ENSG00000100591-AHSA1/tissue/hippocampal+formation#imid_549633
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000100591-AHSA1/tissue/colon#imid_549558
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000100591-AHSA1/tissue/liver#imid_549559
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA000903; green = AHSA1, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA000903; green = AHSA1, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for AHSA1 IHC & IF
🧪Run fallopian tube first and expect staining in its glandular cells (HPA: High in fallopian tube glandular cells); use salivary gland glandular cells as the negative tissue comparator (HPA: Not detected in salivary gland glandular cells). On the positive slide, any unstained neighboring cells can provide a background reference, but the supplied HPA row does not identify a specific AHSA1-negative cell type within fallopian tube (HPA: fallopian tube glandular cells High).
Positive control tissue: Fallopian tube (Glandular cells, HPA High)
Negative control tissue: Salivary gland (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show AHSA1 in A-431, U-251MG, U2OS, with annotated localisation: Cytosol (supported) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only slide and a concentration-matched rabbit isotype control (selected-SKU IHC caption: rabbit primary antibody), plus AHSA1 knockout material or an antibody peptide-block control where available. Quench endogenous peroxidase and check for tissue pigment before interpreting DAB signal (selected-SKU IHC caption: peroxidase detection with DAB).
⚠️Feasibility: No target-specific fixation window or fixation effect is reported in the supplied evidence, and the fixative used for the selected-SKU paraffin-section image is unreported (selected-SKU IHC caption: fixative not stated). Heat retrieval in EDTA at pH 8.0 is a documented IHC condition, but a requirement for that retrieval method has not been established (selected-SKU IHC caption: EDTA pH 8.0). The evidence does not establish that frozen sections or IF are easier; IF images support a cytosolic pattern in A-431, U-251MG and U2OS cells, while the supplied fallopian tube IHC row identifies glandular cells as the expected positive compartment (HPA: Cytosol supported; HPA: fallopian tube glandular cells High).
HPA tissue IHC evidence for AHSA1
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Supported — Medium consistency between antibody staining and RNA expression data.). Rows are taken directly from the HPA tissue chart — click any row's HPA link to view the source.
Positive expression · recommended positive controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Fallopian tube | Glandular cells | High | Protein (IHC) | HPA → |
| Testis | Elongated or late spermatids | High | Protein (IHC) | HPA → |
| Tonsil | Germinal center cells | High | Protein (IHC) | HPA → |
| Adipose tissue | Adipocytes | Medium | Protein (IHC) | HPA → |
| Adrenal gland | Glandular cells | Medium | Protein (IHC) | HPA → |
Section 3
Advanced AHSA1 IHC Tips
Use cytoplasmic staining as the main readout for AHSA1 in paraffin sections (HPA: cytoplasmic expression in most tissues; UniProt O95433: cytosol).
Which retrieval conditions should I try first for AHSA1 in paraffin sections?
Start with heat-mediated retrieval in EDTA at pH 8.0 for AHSA1 paraffin sections (datasheet A05733-2). The documented stain used 2 μg/ml primary antibody overnight at 4°C after retrieval, so keep antibody concentration and incubation consistent while assessing retrieval (caption A05733-2). Compare a retrieved section with an otherwise matched section lacking retrieval, and inspect both target cells and section edges for gain in staining or tissue damage (standard IHC practice). If staining remains weak, adjust heating duration in a small controlled series before trying a different buffer; the caption does not report a heating time (caption A05733-2; standard IHC practice).
How should I assess whether fixation is suppressing AHSA1 staining?
Target-specific fixation sensitivity is unknown: the documented AHSA1 tissue section was paraffin-embedded, but its fixative was not reported (caption A05733-2). Record fixative, time in fixative, tissue thickness, and processing history for each specimen before comparing staining intensity across cases (standard IHC practice). Hold EDTA retrieval at pH 8.0 and the documented 2 μg/ml primary incubation constant while comparing specimens with different known fixation histories (datasheet A05733-2; caption A05733-2; standard IHC practice). Uneven staining across a section or loss of morphology warrants a processing review, but neither HPA tissue patterns nor AHSA1 topology establishes a fixation-specific effect (standard IHC practice; HPA: tissue IHC; UniProt O95433: topology).
What staining compartment should count as plausible AHSA1 signal?
Score cytoplasmic staining first: HPA reports cytoplasmic expression in most tissues and supported cytosolic localisation in cell images (HPA: tissue IHC; HPA: subcellular). UniProt places AHSA1 in the cytosol and notes that interaction with the endoplasmic reticulum may be transient, so a perinuclear accent can be examined without treating it as a required pattern (UniProt O95433: subcellular location). AHSA1 has no annotated transmembrane segment, making a crisp cell-surface-only pattern a reason to check specificity and detection artefacts (UniProt O95433: topology; standard IHC practice). Compare staining within defined cell types and inspect a matched negative control before interpreting nuclear or surface-dominant signal as AHSA1 (standard IHC practice).
Could AHSA1 isoforms or modifications change what this antibody detects?
AHSA1 has 2 annotated isoforms, but the supplied IHC caption does not define this antibody's epitope or isoform coverage (UniProt O95433: isoforms; caption A05733-2). The record lists modified residues including phosphoserine 193 and phosphotyrosine 223; their effect on tissue staining cannot be inferred without an epitope map (UniProt O95433: modified residues; standard IHC interpretation). When staining differs across specimens, compare sections under the same retrieval and detection conditions before assigning the difference to an isoform or modification (standard IHC practice). If isoform specificity matters, obtain the antibody's mapped epitope and compare it with the annotated isoform sequences, then validate the staining assignment independently (standard IHC practice).
How can IF help check an AHSA1 IHC localisation call?
Use IF as a separate localisation check for an ambiguous chromogenic IHC pattern; the supplied product caption documents paraffin-section IHC with DAB, not an IF protocol (caption A05733-2). Multiplex AHSA1 with an epithelial marker such as cytokeratin when examining glandular cells, which HPA reports as AHSA1-positive in fallopian tube and breast (HPA: tissue IHC; standard IF practice). Select spectrally separated dyes and consider a far-red AHSA1 channel where tissue autofluorescence complicates shorter wavelengths, with single-stain controls to assess bleed-through (standard IF practice). Because AHSA1 is cytosolic and lacks a transmembrane segment, permeabilise the plasma membrane for intracellular access and verify that treatment preserves cell boundaries (UniProt O95433: subcellular location and topology; standard IF practice).
What should I check when AHSA1 DAB staining looks diffuse or excessive?
First compare the stained section with a no-primary control to identify signal from the detection system, and check whether excess DAB obscures cellular boundaries (standard IHC practice). The documented workflow blocked with 10% goat serum, used 2 μg/ml primary overnight at 4°C, and detected with a peroxidase secondary and DAB (caption A05733-2). If background persists, titrate primary concentration and development time while keeping retrieval and tissue processing matched across the comparison (standard IHC practice). Include a peroxidase block as a general chromogenic IHC step, and assess tissue edges separately because edge staining can exaggerate apparent cytoplasmic signal (standard IHC practice).
How should I quantify AHSA1 staining across paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and cytoplasmic compartment before scoring, since HPA describes AHSA1 as cytoplasmic in most tissues and reports different levels across cell types (HPA: tissue IHC). For chromogenic IHC, record the percentage of positive target cells and an intensity score, then calculate an H-score on a consistent 0–300 scale if that scoring system fits the study (standard IHC practice). Normalise counts to the number of evaluable target cells, or stained area to evaluable tissue area, and exclude folds, necrosis, and section edges using prespecified rules (standard IHC practice). Keep retrieval, DAB development, imaging settings, and scoring thresholds consistent, and report cell-type-specific results rather than pooling unlike compartments (standard IHC practice).
How can I distinguish convincing AHSA1 staining from artefact?
A convincing result places signal mainly in the cytoplasm of morphologically intact cells, consistent with HPA tissue staining and supported cytosolic localisation (HPA: tissue IHC; HPA: subcellular). Compare expected cell populations with internal contrasts: HPA reports high staining in fallopian tube glandular cells but no detection in salivary gland glandular cells, although its tissue IHC reliability is only Supported (HPA: tissue IHC). Treat isolated nuclear or crisp membrane-only staining cautiously because AHSA1 is assigned to the cytosol and has no transmembrane segment (UniProt O95433: subcellular location and topology). Review edge effects, necrotic regions, and no-primary controls; residual endogenous peroxidase can produce DAB signal unrelated to AHSA1 (standard IHC practice).
Boster reagents
Best AHSA1 / Activator of 90 kDa heat shock protein ATPase homolog 1 IHC Antibodies
A05733-2 has IHC images from paraffin-embedded human carcinoma sections and an IF image from A431 cells (catalog image captions); listed reactivity covers human, mouse, and rat (catalog: reactivity).
Real IHC data
Anti-AHA1/AHSA1 Antibody ®
Cat # A05733-2
A05733-2 is listed for IHC, ICC, and IF in human, mouse, and rat (catalog: applications and reactivity). Its IHC captions show human breast infiltrating ductal carcinoma, colorectal adenocarcinoma, endometrial cancer, and laryngeal squamous cell carcinoma sections; its IF caption shows A431 cells (A05733-2 image captions).
Which to pick: For tissue IHC, choose A05733-2: its captions document paraffin-embedded sections with EDTA retrieval at pH 8.0 and 2 μg/ml primary antibody (A05733-2 IHC captions). For IF/ICC, the same SKU has an A431 cell IF image using 5 μg/ml primary antibody (A05733-2 IF caption). For cross-species work, A05733-2 lists human, mouse, and rat reactivity; clone identity and the IHC section fixative are unreported (catalog: reactivity and clone; A05733-2 IHC captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry O95433 (AHSA1_HUMAN, Activator of 90 kDa heat shock protein ATPase homolog 1).
- Human Protein Atlas. AHSA1 tissue IHC expression (reliability: Supported).
- Human Protein Atlas. AHSA1 subcellular location (ICC-IF): Localized to the cytosol..
- Human Protein Atlas. AHSA1 antibody validation summary (2 antibodies).
- AHSA1 as a prognostic biomarker and potential immunotherapeutic target in HNSCC: integrative bulk RNA-seq, scRNA-seq analyses and experimental validation. BMC cancer 2025 — PMC12519801.
- The Prognostic and Immunotherapeutic Significance of AHSA1 in Pan-Cancer, and Its Relationship With the Proliferation and Metastasis of Hepatocellular Carcinoma. Frontiers in immunology 2022 — PMC9226343.
- The prognostic value of immune escape-related genes in lung adenocarcinoma. Translational cancer research 2024 — PMC11231773.
- AHSA1 is a promising therapeutic target for cellular proliferation and proteasome inhibitor resistance in multiple myeloma. Journal of experimental & clinical cancer research : CR 2022 — PMC8734095.
- PubMed PMID:10931946 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
- PubMed PMID:12508121 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for AHSA1 (Activator of 90 kDa heat shock protein ATPase homolog 1). Expected localisation Cytoplasmic staining in most tissues (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A05733-2). Validated IHC antibody: A05733-2 at 2-5 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Fallopian tube (Glandular cells); negative Salivary gland. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human breast infiltrating ductal carcinoma tissue; fixative not specified…; 2. Fixation: Image fixative and duration unreported (datasheet A05733-2); 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 A05733-2); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A05733-2); 8. Primary antibody: Rabbit anti-AHSA1, 2-5 μg/ml (datasheet A05733-2); 9. Primary incubation: Overnight at 4 °C (datasheet A05733-2). Watch for: Fixation: Keep fixation consistent across sections (standard IHC practice; not target-specific); Caveat: Staining has medium consistency with RNA data (HPA tissue IHC); Isoform / epitope: 2 isoforms; epitope differences unreported (UniProt); Regulation: Expression regulation unreported (UniProt). Evidence: UniProt O95433, HPA tissue IHC · Supported, HPA ICC-IF · cytosol., Datasheet A05733-2, PMC12519801. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/AHSA1/immunohistochemistry-ihc-guide.