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- Table of Contents
ITGA8 / Integrin alpha-8 · Western blot design guide
Design a Western Blot for ITGA8
Source-linked ITGA8 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-ITGA8 WB antibodies. Everything you need to plan the experiment before you commit precious samples.
Evidence assembled September 2026 · For research use; verify linked source records and product datasheet before use
ITGA8 Western Blot Experimental Design Guide
Expected bands, source-linked protocol options, controls and antibodies — the at-a-glance facts below, then the full design guide.
Must know before running
| Expected band | ~117.5 kDa | |
| Observed band | ~180 kDa | |
| Gel | 5–20% (catalog A06636-2) | |
| Positive control | Kidney (IHC candidate; verify WB) | |
| Negative control | Adipose tissue (IHC candidate; verify WB) |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Glycosylated + Cleaved | |
| Caveat | Modification-state controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 1 isoform(s) |
Section 1
Source-Linked ITGA8 Western Blot Protocol Options
The A06636-2 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human HEK293 (catalog A06636-2) |
| Gel % | 5–20% (catalog A06636-2) |
| Load | 30 ug; reducing conditions (catalog A06636-2) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog A06636-2) |
| Membrane | nitrocellulose membrane (catalog A06636-2) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog A06636-2) |
| Primary antibody | A06636-2 · 0.5 μg/mL (catalog A06636-2) |
| Primary incubation | overnight at 4°C (catalog A06636-2) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog A06636-2) |
| Secondary incubation | 1.5 hour at RT (catalog A06636-2) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog A06636-2) |
| Detection | ECL (catalog A06636-2) |
Section 2
What Is the Expected ITGA8 Western Blot Band Size?
ITGA8 is predicted at 117.5 kDa, while a reducing blot shows approximately 180 kDa; the cause of this difference is not established.
| Band near 180 kDa | Empirical ITGA8 band in a reducing whole-cell lysate; its difference from the predicted mass is unexplained |
| Band near 117.5 kDa | Migration near the predicted precursor mass; confirm identity independently |
| Lower band after reduction | Could represent a separated heavy or light chain of ITGA8 |
| Slightly lower band | Could reflect removal of the 1–38 signal peptide |
| Diffuse band | Could reflect variation in N-linked glycosylation; the listed sites alone do not establish a smear |
💡Expected ITGA8 appearanceITGA8 has a predicted precursor mass of 117.5 kDa, while a reducing whole-cell blot reports a band near 180 kDa; the cause of that difference is unestablished, so confirm identity with appropriate controls.
| Predicted precursor mass | 117.5 kDa from the UniProt sequence; the observed band is near 180 kDa |
| N-linked glycosylation at Asn-81 | May affect apparent migration, but its contribution to the 180 kDa band is unestablished |
| N-linked glycosylation at Asn-122 | May affect apparent migration, but no site-specific size shift is supplied |
| Signal peptide at residues 1–38 | Removal makes the mature chain smaller than the precursor; no resulting band size is supplied |
| Disulfide-linked heavy and light chains | Reduction can separate the chains; their individual band sizes are not supplied |
| Situation | Likely cause | Next action |
|---|---|---|
| Band higher than expected | The reported 180 kDa band exceeds the 117.5 kDa predicted precursor mass; the cause is unestablished | Compare with the reported reducing lysate blot and confirm identity using an independent antibody or ITGA8 depletion |
| Band lower than expected | Signal peptide removal or separation of the heavy and light chains may lower band position | Check reducing conditions and confirm the band with an independent ITGA8 antibody |
| Broad smear instead of sharp band | Variable N-linked glycosylation is possible, but a smear is not demonstrated by the listed sites | Compare untreated and deglycosylated samples and verify that the signal is ITGA8 |
| Multiple bands | Disulfide-linked heavy and light chains can separate under reduction | Compare reducing and nonreducing lanes and confirm bands with independent ITGA8 detection |
| Weak or no signal | ITGA8 is a cell-membrane protein and may be poorly recovered in a lysate preparation | Check membrane-protein recovery and include a confirmed ITGA8-positive sample |
| Fragments below expected size | The alpha subunit consists of heavy and light chains linked by a disulfide bond | Check reduction and sample handling; verify fragment identity with an independent antibody |
Experiment evidence
Western blot images showing ITGA8 band
A06636-2 · ITGA8
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for ITGA8 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for ITGA8 in Western blot, you can use kidney tissue lysate.
Positive control: Kidney (IHC candidate; verify WB)
Negative control: Adipose tissue (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside the samples.
⚠️Feasibility: As a membrane protein, ITGA8 may be easier to detect in a membrane-enriched fraction.
HPA tissue expression evidence for ITGA8
Comprehensive Human Protein Atlas IHC scoring per tissue. Rows are taken directly from the HPA tissue chart — click any row's HPA link to view the source.
Higher expression tissues · candidate positive controls from IHC
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Kidney | cells in glomeruli | High | Protein (IHC) | HPA → |
Lower expression tissues · IHC evidence, not confirmed WB-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 Western Blot Tips
Deeper troubleshooting and optimisation questions for ITGA8, answered from its protein features.
How should ITGA8 band migration be interpreted?
Band shift · Use the separately labelled calculated mass and catalog-observed evidence above. A sequence annotation does not establish an observed migration shift. Verify target identity with orthogonal controls.
Do ITGA8 isoforms explain multiple bands?
Isoforms · The supplied record lists one isoform and no alternative sequence. It therefore provides no isoform-based explanation for multiple bands. Assess the listed glycosylation and heavy/light-chain features when investigating them.
Which ITGA8 glycosylation sites matter when assessing a band shift?
PTM · UniProt lists N-linked sites at positions 81, 122, 177, 239, 302, 311, 504, 601, 605, 719, 737, 753, 780, 896, 923, and 1005. These are UniProt coordinates; antibody or paper numbering may differ. The site list alone cannot predict a visible shift.
Does this guide establish induction of ITGA8?
Induction · No general induction response is established by this guide. A pathway or gene-set association is not evidence of induction in a particular specimen. Verify the relevant treatment and control in a target-specific experiment.
What transfer method to use for ITGA8 Western blot?
Transfer · ITGA8 is a single-pass membrane protein with an observed apparent band near 180 kDa. Choose and verify transfer conditions that recover a band in that high-mass region; check the post-transfer gel and membrane before interpreting a weak or absent signal.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A06636-2 datasheet where specified. Otherwise compare 5% milk or 5% BSA in TBST; for a phospho-specific assay start with BSA. Optimize background and specific signal with matched controls.
How should ITGA8 bands be quantified?
Quantitation · Quantify the same defined band across samples and verify that its signal is within the assay's linear range. ITGA8 is a membrane protein, so use consistent sample extraction and loading. If both high-mass and smaller bands appear, report which band was measured.
Why might ITGA8 appear near 180 kDa instead of 117.5 kDa?
Interpretation · 117.5 kDa is the predicted mass, while approximately 180 kDa is the observed apparent band. ITGA8 has a signal peptide at UniProt positions 1–38 and 16 listed N-linked glycosylation sites. These features warrant checking processing and glycosylation, but their presence alone does not establish the cause or size of the difference.
UniProt describes the alpha subunit as heavy and light chains linked by a disulfide bond, and lists cleavage on paired basic residues. When interpreting extra bands, consider whether sample preparation preserves that linkage and which chain the antibody recognizes. The supplied features do not specify a cleavage position or chain masses.
Check whether each band is compatible with the observed approximately 180 kDa species, glycosylation, or the disulfide-linked heavy and light chains. Antibody recognition and sample preparation affect which species can be seen. The listed features suggest possibilities; they do not identify an unexpected band by themselves.
UniProt describes the alpha subunit as heavy and light chains linked by a disulfide bond, and lists cleavage on paired basic residues. When interpreting extra bands, consider whether sample preparation preserves that linkage and which chain the antibody recognizes. The supplied features do not specify a cleavage position or chain masses.
Check whether each band is compatible with the observed approximately 180 kDa species, glycosylation, or the disulfide-linked heavy and light chains. Antibody recognition and sample preparation affect which species can be seen. The listed features suggest possibilities; they do not identify an unexpected band by themselves.
Boster reagents
ITGA8 Western Blot Antibodies
Catalog antibodies with Western blot application and product-specific WB images. Evaluate suitability with the reported sample, controls and experimental conditions.
Real WB data
Anti-ITGA8 Antibody Picoband®
Cat # A06636-2
The catalog reports one anti-ITGA8 antibody for Western blotting, A06636-2, with human reactivity and a blot of HEK293 whole-cell lysate. The reported band is approximately 180 kDa, versus an expected 117 kDa; evidence for other samples is not supplied.
Which to pick: A06636-2 is the only listed option and has a Western blot image using human HEK293 lysate under reducing conditions. Consider the reported 180 kDa band and 117 kDa expected size when interpreting results.
Source: BosterBio ITGA8 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P53708.
- Human Protein Atlas. ITGA8 tissue expression.
- PMC7475494 — target-verified WB comparison
- PMC9995370 — target-verified WB comparison
- PMC12752560 — target-verified WB comparison