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ADAM8 / Disintegrin and metalloproteinase domain-containing protein 8 · Western blot design guide
Design a Western Blot for ADAM8
Source-linked ADAM8 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-ADAM8 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
ADAM8 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 | ~88.8 kDa | |
| Observed band | ~80 kDa | |
| Gel | 5–20% (catalog A04306-4) | |
| Positive control | Adrenal gland (IHC candidate; verify WB) +4 more | |
| Negative control | Cerebellum (IHC candidate; verify WB) |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Glycosylated + Cleaved | |
| Caveat | Glycosylation-state controls | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 3 isoform(s) |
Section 1
Source-Linked ADAM8 Western Blot Protocol Options
The A04306-4 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human Raji, human Caco-2, rat PC-12 (catalog A04306-4) |
| Gel % | 5–20% (catalog A04306-4) |
| Load | 30 ug; reducing conditions (catalog A04306-4) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog A04306-4) |
| Membrane | nitrocellulose membrane (catalog A04306-4) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog A04306-4) |
| Primary antibody | A04306-4 · 0.5 μg/mL (catalog A04306-4) |
| Primary incubation | overnight at 4°C (catalog A04306-4) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog A04306-4) |
| Secondary incubation | 1.5 hour at RT (catalog A04306-4) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog A04306-4) |
| Detection | ECL (catalog A04306-4) |
Section 2
What Is the Expected ADAM8 Western Blot Band Size?
ADAM8 is predicted at 88.8 kDa and observed near 80 kDa in reducing lysates; the cause of the difference is not established.
| Band near 80 kDa | Empirical ADAM8 band in reducing whole-cell lysates; confirm identity with an orthogonal antibody or ADAM8 depletion |
| Band near 88.8 kDa | Could reflect migration near the calculated full-length precursor mass; identity requires confirmation |
| Band below the full-length precursor position | Signal-peptide cleavage could reduce size, but its contribution to the observed 80 kDa band is unestablished |
| Additional bands at different positions | ADAM8 has isoforms 1, 2, and 3, though distinct migration has not been demonstrated |
| Diffuse or higher band | Variation in N-linked glycosylation is possible at Asn67, Asn91, Asn436, and Asn612; a visible effect is unproven |
💡Expected ADAM8 appearanceADAM8 has a calculated full-length mass of 88.8 kDa and an empirical band near 80 kDa in reducing lysates; the difference is unexplained, so confirm band identity with depletion or an orthogonal antibody.
| Calculated full-length mass | 88.8 kDa is the sequence-based reference; the empirical band is near 80 kDa |
| N-linked glycosylation at Asn67, Asn91, Asn436, and Asn612 | Could increase or vary apparent mass, but site annotations alone establish no visible shift |
| Signal peptide at residues 1–16 | Cleavage makes the mature chain smaller than the full-length precursor; its contribution to the 80 kDa band is unestablished |
| Splice isoforms 1, 2, and 3 | May differ in size; their relative masses and distinct band positions are not supplied |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | ADAM8 is a single-pass membrane protein and may be poorly recovered in a preparation | Check membrane-protein extraction and include a positive-control lysate |
| Band higher than expected | N-linked glycosylation could affect migration, but the cause of a higher band is unconfirmed | Compare matched samples before and after N-glycan removal and verify identity |
| Band lower than expected | The observed 80 kDa band is below the calculated 88.8 kDa full-length mass; the cause is unestablished | Confirm with ADAM8 depletion or an orthogonal antibody |
| Broad smear instead of sharp band | Variable N-linked glycosylation is possible but has not been shown to cause a smear | Compare matched samples after N-glycan removal and check sample integrity |
| Multiple bands | Isoforms 1, 2, and 3 exist, but their migration patterns are not supplied | Use ADAM8 depletion or an orthogonal antibody to identify specific bands |
| Weak or no signal | Membrane-protein recovery or low ADAM8 abundance in the tested lysate may limit detection | Check extraction and loading with a positive-control lysate |
Experiment evidence
Western blot images showing ADAM8 band
A04306-4 · ADAM8
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for ADAM8 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for ADAM8 in Western blot, you can use adrenal gland tissue.
Positive control: Adrenal gland (IHC candidate; verify WB)
Negative control: Cerebellum (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside the samples.
⚠️Feasibility: As a membrane protein, ADAM8 may be easier to detect in membrane-enriched lysates.
HPA tissue expression evidence for ADAM8
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 |
|---|---|---|---|---|
| Adrenal gland | glandular cells | Medium | Protein (IHC) | HPA → |
| Appendix | glandular cells | Medium | Protein (IHC) | HPA → |
| Bone marrow | hematopoietic cells | Medium | Protein (IHC) | HPA → |
| Cerebral cortex | neuronal cells | Medium | Protein (IHC) | HPA → |
| Duodenum | glandular cells | Medium | Protein (IHC) | HPA → |
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Cerebellum | cells in granular layer | Not detected | Protein (IHC) | HPA → |
| Esophagus | squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
| Heart muscle | cardiomyocytes | Not detected | Protein (IHC) | HPA → |
| Liver | cholangiocytes | Not detected | Protein (IHC) | HPA → |
| Oral mucosa | squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced ADAM8 Western Blot Tips
Deeper troubleshooting and optimisation questions for ADAM8, answered from its protein features.
How should ADAM8 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.
Which ADAM8 isoforms could produce different bands?
Isoforms · UniProt lists isoforms 1, 2, and 3. Relative to the canonical sequence, isoform 2 replaces residues 16–92 and lacks 93–131, 596–621, and 774–799. Isoform 3 replaces 650–742 and lacks 743–824. Check whether the antibody recognizes a region retained in the isoform you intend to measure.
Which glycosylation sites matter when interpreting ADAM8 bands?
PTM · The canonical sequence has N-linked glycosylation sites at UniProt positions 67, 91, 436, and 612. Isoform 2 changes the sequence covering 67 and 91 and lacks the segment containing 612; isoform 3 retains all four listed positions. These are canonical UniProt coordinates, which may differ from antibody or paper numbering. Site annotations alone do not prove a visible band shift.
Does this guide establish induction of ADAM8?
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.
How should transfer be checked for ADAM8?
Transfer · Standard workflow guidance: verify transfer efficiency for the intended target size before interpreting a weak signal. Use total-protein assessment and optimize transfer for the membrane, gel and apparatus; the labelled catalog values take precedence.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A04306-4 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 I quantify ADAM8 across samples?
Quantitation · Decide which isoform or band you are measuring before comparing samples. For coverage of all three listed isoforms, consider an antibody recognizing an unchanged region such as canonical residues 132–435, then verify its band assignment. Report the same defined band or bands consistently; the supplied features do not establish that every ADAM8 band represents the same form.
Why might ADAM8 appear near 80 kDa rather than 88.8 kDa?
Interpretation · The 88.8 kDa prediction refers to the canonical sequence, while the reported band is approximately 80 kDa. ADAM8 has a signal peptide at residues 1–16, three isoforms, and four N-linked glycosylation sites. These features warrant checking which form the antibody detects, but they do not establish the cause of the mass difference.
UniProt lists 12 disulfide bonds. Compare samples prepared under consistent reducing conditions when assessing band patterns, since disulfide-dependent structure can affect migration. The feature count alone cannot predict the direction or size of a change.
First compare each band with the approximately 80 kDa reported band and the 88.8 kDa canonical prediction. Then check the antibody's target region against the listed isoform changes, signal peptide at 1–16, and glycosylation sites at 67, 91, 436, and 612. These features suggest possibilities, but band position alone cannot identify an isoform or modification.
UniProt lists 12 disulfide bonds. Compare samples prepared under consistent reducing conditions when assessing band patterns, since disulfide-dependent structure can affect migration. The feature count alone cannot predict the direction or size of a change.
First compare each band with the approximately 80 kDa reported band and the 88.8 kDa canonical prediction. Then check the antibody's target region against the listed isoform changes, signal peptide at 1–16, and glycosylation sites at 67, 91, 436, and 612. These features suggest possibilities, but band position alone cannot identify an isoform or modification.
Boster reagents
ADAM8 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-MS2/ADAM8 Antibody Picoband®
Cat # A04306-4
The catalog reports one anti-ADAM8 antibody, A04306-4, with reported human and rat reactivity. Its WB image shows an approximately 80 kDa band in human Raji and Caco-2 and rat PC-12 whole cell lysates. No publication evidence was supplied.
Which to pick: A04306-4 is the only listed option. Its WB image documents the reported band in human Raji and Caco-2 and rat PC-12 lysates; choose it when those tested contexts and the stated human or rat reactivity fit your experiment.
Source: BosterBio ADAM8 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P78325.
- Human Protein Atlas. ADAM8 tissue expression.
- PMC8875419 — target-verified WB comparison
- PMC4141088 — target-verified WB comparison
- PMC11481351 — target-verified WB comparison