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ITGA8 / Integrin alpha-8 · IHC design guide
Design Immunohistochemistry for ITGA8
Plan chromogenic ITGA8 IHC on paraffin sections using kidney glomeruli as a high-staining reference (HPA tissue IHC) and the IHC-validated antibody at 1:50–1:100 (datasheet: A06636-1 IHC-P). Assess the observed vascular cytoplasmic staining (HPA tissue IHC) alongside ITGA8’s expected cell membrane location (UniProt).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
ITGA8 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 | Vascular cytoplasm observed (HPA tissue IHC); cell membrane expected (UniProt) | |
| Staining pattern | Cytoplasmic staining mainly in blood vessels; high in glomerular cells (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen) | |
| Positive control | Kidney | |
| Negative control | Adipose tissue+4 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Formalin-fixed paraffin sections are documented (datasheet: A06636-1 IHC-P). (selected-SKU IHC image A06636-1) | |
| Caveat | Vascular staining may complicate glomerular scoring (HPA tissue IHC) | |
| Regulation | Expression regulation not specified (UniProt) | |
| Isoform / epitope | 0 isoforms; cleavage and extracellular vs cytoplasmic epitope matter (UniProt) |
Section 1
Recommended ITGA8 IHC & IF Protocols
The catalog antibody’s IHC-P protocol is accompanied by one published IHC workflow for human samples (PMC6975017: Methods).
| Sample | FFPE human kidney tissue (datasheet A06636-1) |
| Fixation | Image formalin-fixed; duration unreported (datasheet A06636-1); 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-ITGA8, 1:50-1:100 (datasheet A06636-1) |
| 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 | ITGA8-positive staining in cells in glomeruli of kidney (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression mainly in blood vessels. No signal in the no-primary control. |
💡Decision noteStart with citrate pH 6.0 HIER at 95–98 °C for 20 min (page antigen retrieval); PMC6975017 does not specify retrieval conditions.
Section 2
What Is the Expected ITGA8 Staining Pattern?
ITGA8 is a cell membrane protein with an extracellular region and short cytoplasmic tail (UniProt P53708 topology). In tissue IHC, HPA reports mainly cytoplasmic staining in blood vessels and high staining in kidney glomerular cells (HPA tissue IHC, Enhanced reliability). Read the vessel-associated signal alongside the expected membrane localisation; HPA's tissue description does not establish that every cytoplasmic signal represents intracellular ITGA8.
| Vessel-associated staining and strong staining in kidney glomerular cells. | This matches HPA's reported distribution: mainly cytoplasmic expression in blood vessels and high staining in glomerular cells (HPA tissue IHC). Record which structures stain rather than scoring the section as uniformly positive (general IHC practice). |
| Staining outlines cells or lies close to their borders within a positive structure. | A peripheral pattern is consistent with ITGA8 at the cell membrane (UniProt P53708 topology; HPA ICC-IF: approved plasma membrane location). HPA describes its tissue IHC signal as mainly cytoplasmic, so compartment alone cannot decide whether an IHC result is valid (HPA tissue IHC). |
| Strong nuclear staining dominates, with little staining in vessels or glomerular cells. | Nuclear localisation conflicts with the reported membrane location (UniProt P53708 topology; HPA ICC-IF). Treat it as suspect, then compare with a reagent control and a kidney positive section before calling it ITGA8 (general IHC practice; HPA tissue IHC: high in glomerular cells). |
| Respiratory epithelium or adipocytes stain strongly while expected structures are weak. | HPA reports ITGA8 as not detected in bronchial respiratory epithelial cells and adipocytes (HPA tissue IHC). Consider off-target staining or endogenous detection activity; check cell identity and controls before interpreting those cells as positive (general IHC practice). |
| Broad, low-level colour covers cells and extracellular areas without a clear structural pattern. | This does not resemble HPA's vessel-focused tissue profile (HPA tissue IHC). Diffuse signal can reflect background from detection or insufficient blocking (general IHC practice); compare a reagent control and inspect whether glomerular cells remain distinguishable (HPA tissue IHC: high in glomerular cells). |
💡Expected ITGA8 appearanceCall an IHC section positive when high signal identifies kidney glomerular cells or staining follows blood vessels (HPA tissue IHC), with peripheral signal compatible with membrane ITGA8 (UniProt P53708 topology); dominant nuclear or widespread staining in HPA-negative cells is suspect (HPA ICC-IF; HPA tissue IHC; general IHC practice).
| Tissue and cell choice | Kidney glomerular cells offer an HPA-reported high reference, while the overall tissue profile is mainly vessel-associated (HPA tissue IHC). UniProt lists mesenchymal populations, including kidney mesangial and smooth muscle cells, but HPA's label 'cells in glomeruli' does not identify a precise cell subtype (UniProt P53708 tissue specificity; HPA tissue IHC). |
| Compartment and epitope orientation | The extracellular region spans residues 39–1012, the transmembrane segment 1013–1033, and the cytoplasmic tail 1034–1063 (UniProt P53708 topology). The antibody epitope is unspecified here; these coordinates cannot predict its staining pattern or retrieval needs. |
| Processing and glycosylation | UniProt describes disulfide-linked heavy and light chains and 16 glycosylation sites (UniProt P53708 processing and glycosylation). The supplied sources do not show how these features affect this antibody's IHC signal; do not infer shedding, loss of staining, or a required retrieval method. |
| Antibody evidence and negative comparators | HPA lists antibody HPA003432 as IHC Enhanced; its tissue profile has Enhanced reliability from staining and RNA consistency (HPA antibodies; HPA tissue IHC). HPA reports no detection in bronchial respiratory epithelial cells and adipocytes, useful cell-level comparators rather than proof that an entire section must be blank (HPA tissue IHC). |
| IF/ICC question: what localisation is expected? | Answer: plasma membrane is the approved ICC-IF location, with images listed for HEK293 and U2OS (HPA subcellular). This supports a localisation check in the separate IF/ICC guide; it does not prescribe an IF protocol or override the mainly cytoplasmic vessel description for tissue IHC (HPA tissue IHC). |
| Fixation and antigen retrieval | No target-specific fixation sensitivity or retrieval condition is supplied by UniProt or HPA. If optimisation is needed, compare retrieval conditions using the same positive tissue and controls; this is general IHC practice, not an ITGA8-specific finding. |
| Situation | Likely cause | Next action |
|---|---|---|
| No signal in kidney glomerular cells. | The section may be uninformative, or the IHC workflow may have failed; HPA reports high glomerular-cell staining (HPA tissue IHC). The supplied sources do not identify a target-specific failure mechanism. | Check tissue preservation and detection controls, then compare routine retrieval and antibody conditions on a kidney section (general IHC practice). Score the expected glomerular cells, not the kidney section as a whole (HPA tissue IHC). |
| Signal appears mainly nuclear. | Nuclear dominance conflicts with membrane localisation (UniProt P53708 topology; HPA ICC-IF). Its precise cause cannot be assigned from localisation alone. | Inspect a reagent control and the distribution in vessels and glomerular cells; review detection background before assigning the nuclear colour to ITGA8 (general IHC practice; HPA tissue IHC). |
| Strong staining appears in bronchial respiratory epithelial cells or adipocytes. | Those cells are listed as not detected by HPA tissue IHC (HPA tissue IHC); possible causes include off-target binding or endogenous detection activity (general IHC practice). | Confirm the cell type, compare a reagent control, and check whether the expected vessel or glomerular pattern is also present (general IHC practice; HPA tissue IHC). |
| Diffuse colour obscures vessel and glomerular-cell boundaries. | Non-specific reagent binding or detection background can reduce contrast (general IHC practice). HPA's tissue description calls for a mainly vessel-associated pattern, with high signal in glomerular cells (HPA tissue IHC). | Inspect reagent controls, blocking and wash steps, and detection exposure; adjust the general IHC workflow until structures can be distinguished (general IHC practice). |
| Vessels stain, but the signal looks cytoplasmic rather than sharply peripheral. | HPA explicitly describes tissue IHC staining as mainly cytoplasmic in blood vessels, despite approved plasma membrane localisation by ICC-IF and membrane topology (HPA tissue IHC; HPA ICC-IF; UniProt P53708 topology). | Judge the vessel-associated pattern together with controls and glomerular staining; document the compartment seen rather than discarding a tissue pattern solely because it lacks a crisp membrane outline (HPA tissue IHC; general IHC practice). |
| Different retrieval conditions give different staining intensity. | The supplied UniProt and HPA evidence does not establish an ITGA8-specific fixation or retrieval effect. Variation between preparation conditions must be assessed experimentally (general IHC practice). | Hold tissue, antibody and detection conditions constant while comparing retrieval conditions; retain the condition that gives interpretable glomerular and vessel signal with acceptable controls (HPA tissue IHC; general IHC practice). |
Experiment evidence
IHC & IF images showing ITGA8 staining
A06636-1 · IHCITGA8 Antibody (C-term) immunohistochemistry analysis in formalin fixed and paraffin embedded human kidney tissue followed by peroxidase con
A06636-2 · IFIF analysis of ITGA8 using anti-ITGA8 antibody (A06636-2). ITGA8 was detected in an immunocytochemical section of A431 cells. Enzyme antigen
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000077943-ITGA8/tissue/cerebral+cortex#imid_1256772
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000077943-ITGA8/tissue/colon#imid_1256714
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000077943-ITGA8/tissue/liver#imid_1256699
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000077943-ITGA8/tissue/kidney#imid_1256716
HPA · ICC-IF HEK293Human Protein Atlas immunofluorescence, HEK293, antibody HPA003432; green = ITGA8, blue = nucleus, red = microtubules.
HPA · ICC-IF HEK293Human Protein Atlas immunofluorescence, HEK293, antibody HPA003432; green = ITGA8, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for ITGA8 IHC & IF
🧪Run kidney first: cells in glomeruli should stain (HPA: High in kidney cells in glomeruli). Use adipose tissue adipocytes as the negative tissue (HPA: Not detected in adipocytes); on the kidney slide, use cells without specific staining as internal negatives, without assuming that all nonglomerular cells lack ITGA8.
Positive control tissue: Kidney (Cells in glomeruli, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show ITGA8 in HEK293, U2OS, with annotated localisation: Plasma membrane (approved) (HPA subcellular).
Technical controls: Include a no-primary (secondary-only) control, an isotype control matched to the primary antibody’s host species and clonality, and, if available, ITGA8-knockout tissue or a peptide-block control (standard IHC practice). Quench endogenous peroxidase for chromogenic kidney IHC and assess endogenous biotin if using biotin-based detection; check autofluorescence for IF (standard IHC/IF practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported in the supplied evidence, although the selected antibody caption explicitly describes formalin-fixed, paraffin-embedded human kidney with peroxidase/DAB detection (A06636-1 tissue-IHC caption). Retrieval dependence is unreported, so compare retrieval conditions using the same positive and negative controls (standard IHC practice). The supplied evidence does not establish that frozen sections or IF/ICC are easier; HPA reports plasma-membrane localization in ICC-IF images from HEK293 and U2OS cells (HPA subcellular).
HPA tissue IHC evidence for ITGA8
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — High consistency between antibody staining and RNA expression data. Antibody staining in cells/structures not annotated, view images.). 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 |
|---|---|---|---|---|
| Kidney | Cells in glomeruli | 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 → |
| Breast | Adipocytes | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced ITGA8 IHC Tips
Troubleshoot ITGA8 staining in chromogenic IHC by checking retrieval, cell identity, compartment, controls, and scoring before interpreting signal.
How should I retrieve ITGA8 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 minutes (page retrieval specification). Let sections cool in buffer, then compare a kidney section containing glomeruli with the problem section in the same run (standard IHC practice; HPA: High in kidney glomerular cells). If staining remains weak, test a second retrieval condition on adjacent sections while holding antibody concentration and detection constant (standard IHC practice). Score specific cellular signal and tissue damage together, since stronger overall DAB color alone does not establish better ITGA8 detection (standard IHC practice).
Could fixation explain weak or uneven ITGA8 staining?
Target specific fixation sensitivity for ITGA8 is unknown from the supplied evidence; do not assign a preferred fixation duration to this antigen (supplied evidence: no target specific fixation comparison). For paraffin IHC, record fixative, time to fixation, duration, and processing history for each specimen before comparing staining intensity (standard IHC practice). Compare sections with similar documented handling and include a kidney glomerular positive control in each run (standard IHC practice; HPA: High in kidney glomerular cells). If signal varies across a specimen, inspect morphology and section edges, then repeat staining on a comparably processed section before attributing variation to ITGA8 biology (standard IHC practice).
Should I accept cytoplasmic ITGA8 staining in blood vessels?
ITGA8 is a cell membrane protein with an extracellular region at residues 39–1012 and a cytoplasmic tail at 1034–1063 (UniProt P53708 topology). HPA describes tissue IHC as mainly cytoplasmic in blood vessels, while its approved subcellular location is the plasma membrane (HPA: tissue profile; HPA: subcellular). Evaluate whether DAB outlines plausible vascular or glomerular cells, and compare that pattern with the kidney control rather than scoring diffuse tissue color alone (UniProt P53708 tissue specificity; HPA: High in kidney glomerular cells; standard IHC practice). Treat a purely nuclear pattern as suspect and review retrieval, blocking, and antibody controls before interpreting it (UniProt P53708 topology; standard IHC practice).
Can processing or epitope position change what this antibody detects?
The record lists 0 annotated isoforms and describes an alpha 8 heavy chain spanning residues 39–906 linked to a light chain spanning 907–1063 (UniProt P53708 processing and isoforms). The selected IHC image identifies antibody A06636-1 as C terminal, but gives no precise epitope residues (selected hero product tissue IHC caption). Accordingly, establish the mapped epitope before deciding whether cleavage or fixation could affect recognition; a C terminal label alone cannot resolve that question (selected hero product tissue IHC caption; standard IHC practice). Record retrieval and antibody lot alongside staining results when comparing sections, and avoid interpreting a changed pattern as an isoform shift without independent evidence (standard IHC practice; UniProt P53708 isoforms).
How can I check ITGA8 cell identity with multiplex immunofluorescence?
Use IF on a parallel section to pair ITGA8 with a validated marker of the expected mesangial or vascular smooth muscle population, then compare cell identity with the chromogenic IHC pattern (UniProt P53708 tissue specificity; standard IF practice). Choose spectrally separated fluorophores and place the weaker signal in a channel with lower measured tissue autofluorescence (standard IF practice). If the antibody epitope is confirmed within the cytoplasmic tail at residues 1034–1063, optimize mild permeabilisation; an extracellular epitope may be tested without it (UniProt P53708 topology; standard IF practice). Include single stain and secondary only controls, and do not transfer an IF setting directly to DAB scoring (standard IF and IHC practice).
What causes widespread DAB color outside expected ITGA8 positive cells?
First inspect a secondary only control and a section stained without primary antibody to distinguish detection background from primary antibody dependent color (standard IHC practice). In the peroxidase DAB workflow shown for A06636-1, use an endogenous peroxidase block and assess whether residual color follows blood rich areas (selected hero product tissue IHC caption; standard IHC practice). Check that blocking, washes, antibody concentration, and DAB development time are consistent across the run before changing retrieval (standard IHC practice). Interpret broad adipocyte staining cautiously because HPA reports ITGA8 as not detected in adipocytes, while kidney glomerular cells provide a high signal reference (HPA: adipocytes Not detected; HPA: kidney glomerular cells High).
How should I quantify ITGA8 IHC across kidney sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the region and cell population before scoring; for kidney, glomerular cells are an evidence supported target population (HPA: High in kidney glomerular cells). Report the percentage of positive cells with an intensity score or H score, and keep the same positivity threshold and imaging settings across sections (standard IHC practice). Normalize counts to the number of eligible glomerular cells or sampled glomerular area, and report sampled structures and tissue area when using positive cells per mm² (standard IHC practice). Exclude folds, necrosis, and edge staining using prespecified rules, then review ambiguous cytoplasmic color against the expected membrane and vascular context (standard IHC practice; UniProt P53708 topology; HPA: tissue profile).
How do I distinguish genuine ITGA8 staining from artefact?
Look for reproducible signal in plausible mesangial or vascular smooth muscle cells, with a cellular pattern consistent with membrane localization and the reported vascular cytoplasmic IHC profile (UniProt P53708 tissue specificity and topology; HPA: tissue profile). Use kidney glomerular cells as a high signal reference and adipocytes as a reported negative population within appropriate controls (HPA: kidney glomerular cells High; HPA: adipocytes Not detected). Question isolated nuclear staining, section edge color, necrotic debris, or signal reproduced in a no primary control (UniProt P53708 topology; standard IHC practice). Residual endogenous peroxidase can also produce misleading DAB color, so compare blocked controls and morphology before calling a specimen positive (standard IHC practice).
Boster reagents
Best ITGA8 / Integrin alpha-8 IHC Antibodies
Two human-reactive ITGA8 antibodies have image data: one for chromogenic IHC in kidney tissue and one for IF in A431 cells (catalog reactivity; A06636-1 and A06636-2 image captions).
Real IHC data
Anti-ITGA8 Antibody (C-term)
Cat # A06636-1
Real IF data
Anti-ITGA8 Antibody ®
Cat # A06636-2
A06636-1 lists human IHC-P and shows DAB staining of formalin-fixed, paraffin-embedded human kidney (catalog applications/reactivity; A06636-1 image caption). A06636-2 lists human IF/ICC and shows IF staining of A431 cells (catalog applications/reactivity; A06636-2 image caption).
Which to pick: Choose A06636-1 for paraffin-section IHC: it is listed for IHC-P at 1:50–1:100 and has a formalin-fixed, paraffin-embedded kidney image (A06636-1 catalog dilution and image caption). Choose A06636-2 for IF/ICC at 5 μg/ml; its image shows IF in A431 cells, while its application list does not include IHC-P (A06636-2 catalog applications/dilution and image caption). Neither SKU has documented reactivity beyond human, so the payload does not support a cross-species choice (catalog reactivity).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry P53708 (ITA8_HUMAN, Integrin alpha-8).
- Human Protein Atlas. ITGA8 tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. ITGA8 subcellular location (ICC-IF): Localized to the plasma membrane..
- Human Protein Atlas. ITGA8 antibody validation summary (1 antibodies).
- Endosteal integrin-α8⁺ mesenchymal stem cells maintain haematopoietic stem cell function via extracellular matrix-mediated interactions. Nature communications 2026 — PMC13473570.
- ITGA8 suppresses proliferation and metastasis of lung adenocarcinoma through the inhibition of glycolysis. Scientific reports 2026 — PMC13031682.
- Integrin alpha8beta1 is Identified as a Potential CAR-T Target for BCMA-Resistant Relapsed Multiple Myeloma. ImmunoTargets and therapy 2026 — PMC13225147.
- CD248 and integrin alpha-8 are candidate markers for differentiating lung fibroblast subtypes. BMC pulmonary medicine 2020 — PMC6975017.
- PubMed PMID:15164054 — UniProt-cited evidence.
- PubMed PMID:7768999 — UniProt-cited evidence.
- PubMed PMID:10504498 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for ITGA8 (Integrin alpha-8). Expected localisation Vascular cytoplasm observed (HPA tissue IHC); cell membrane expected (UniProt). Antigen retrieval: Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen). Validated IHC antibody: A06636-1 at 1:50-1:100 (the product's own IHC validation image is shown). Controls (HPA): positive Kidney (Cells in glomeruli); negative Adipose tissue. IHC-P workflow on FFPE sections. Protocol steps: 1. Sample: FFPE human kidney tissue (datasheet A06636-1); 2. Fixation: Image formalin-fixed; duration unreported (datasheet A06636-1); 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-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-ITGA8, 1:50-1:100 (datasheet A06636-1); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Formalin-fixed paraffin sections are documented (datasheet: A06636-1 IHC-P). (selected-SKU IHC image A06636-1); Caveat: Vascular staining may complicate glomerular scoring (HPA tissue IHC); Isoform / epitope: 0 isoforms; cleavage and extracellular vs cytoplasmic epitope matter (UniProt); Regulation: Expression regulation not specified (UniProt). Evidence: UniProt P53708, HPA tissue IHC · Enhanced, HPA ICC-IF · plasma membrane., Datasheet A06636-1, PMC6975017. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/ITGA8/immunohistochemistry-ihc-guide.