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- Table of Contents
ELMOD3 / ELMO domain-containing protein 3 · Western blot design guide
Design a Western Blot for ELMOD3
Source-linked ELMOD3 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-ELMOD3 WB antibodies. Everything you need to plan the experiment before you commit precious samples.
Evidence assembled October 2026 · For research use; verify linked source records and product datasheet before use
ELMOD3 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 | ~43 kDa | |
| Gel | 12–15% (standard starting point) | |
| Positive control | Cerebral cortex (IHC candidate; verify WB) +4 more | |
| Negative control | Appendix (IHC candidate; verify WB) |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | — | |
| Caveat | Isoform-specific controls | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 5 isoform(s) |
Section 1
Source-Linked ELMOD3 Western Blot Protocol Options
The A12704 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | extracts of various cell lines, (catalog A12704) |
| Gel % | 12–15% (standard starting point) |
| Load | 20–30 µg total protein per lane; optimize for abundance (standard starting point) |
| Transfer | Standard semi-dry transfer; verify efficiency (standard starting point) |
| Membrane | 0.45 µm PVDF (standard starting point) |
| Blocking | Blocking buffer: 3% nonfat dry milk in TBST (catalog A12704) |
| Primary antibody | A12704 · 1:1000 (catalog A12704) |
| Primary incubation | Overnight at 4 °C (standard starting point) |
| Secondary antibody | Goat Anti-Rabbit IgG, 1:10000 (catalog A12704) |
| Secondary incubation | 1 h at room temperature (standard starting point) |
| Wash | 3 × 5 min in TBST (standard starting point) |
| Detection | ECL (catalog A12704) |
Section 2
What Is the Expected ELMOD3 Western Blot Band Size?
ELMOD3 is predicted at 43 kDa; five isoforms could affect migration, but no empirical band or isoform separation is supplied.
| Band near 43 kDa | Consistent with the predicted ELMOD3 mass; confirm identity with controls |
| Several discrete bands | Could reflect isoforms 1, 2, 3, 5, or 6; their separation is unestablished |
| Band away from 43 kDa | Could represent an isoform, but its mass is unknown |
| Faint or absent band in lysate | ELMOD3 localized to hair-cell projections and the cytoskeleton may be scarce or poorly recovered |
💡Expected ELMOD3 appearanceUniProt predicts 43 kDa, but no empirical band size or isoform migration is supplied; verify a band near 43 kDa with ordinary identity controls.
| UniProt predicted mass | Places the reference band near 43 kDa |
| Isoform 1 | May differ in apparent size from other isoforms; size order is unknown |
| Isoform 2 | May differ in apparent size from other isoforms; size order is unknown |
| Isoform 3 | May differ in apparent size from other isoforms; size order is unknown |
| Isoform 5 | May differ in apparent size from other isoforms; size order is unknown |
| Isoform 6 | May differ in apparent size from other isoforms; size order is unknown |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | The sampled cells may have little ELMOD3, which is reported in hair-cell projections | Check an ELMOD3-expressing reference sample and sample loading |
| Band higher than expected | An isoform or unrelated antibody target is possible; no larger isoform mass is supplied | Compare with ELMOD3 knockdown or knockout material |
| Band lower than expected | An isoform or unrelated antibody target is possible; no smaller isoform mass is supplied | Check whether the band tracks with ELMOD3 knockdown or knockout |
| Multiple bands | Five named splice isoforms could contribute, but distinct migration is unproven | Check antibody epitope coverage and test band identity by ELMOD3 depletion |
| Weak or no signal | Cytoskeletal localization may limit recovery in the tested lysate | Check the insoluble fraction and compare an ELMOD3-expressing reference sample |
Experiment evidence
Western blot images showing ELMOD3 band
A12704 · ELMOD3
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for ELMOD3 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for ELMOD3 in Western blot, you can use cerebral cortex tissue, which HPA rates as highly expressed.
Positive control: Cerebral cortex (IHC candidate; verify WB)
Negative control: Appendix (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain such as Ponceau alongside the samples.
⚠️Feasibility: Tissue controls are feasible, though cytoskeletal localization may affect protein extraction.
HPA tissue expression evidence for ELMOD3
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 |
|---|---|---|---|---|
| Cerebral cortex | endothelial cells | High | Protein (IHC) | HPA → |
| Colon | endothelial cells | High | Protein (IHC) | HPA → |
| Endometrium | cells in endometrial stroma | High | Protein (IHC) | HPA → |
| Lung | alveolar cells | High | Protein (IHC) | HPA → |
| Ovary | ovarian stroma cells | High | Protein (IHC) | HPA → |
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Appendix | glandular cells | Not detected | Protein (IHC) | HPA → |
| Bone marrow | hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Caudate | glial cells | Not detected | Protein (IHC) | HPA → |
| Cerebellum | cells in granular layer | Not detected | Protein (IHC) | HPA → |
| Hippocampus | glial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced ELMOD3 Western Blot Tips
Deeper troubleshooting and optimisation questions for ELMOD3, answered from its protein features.
How should ELMOD3 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.
Could ELMOD3 isoforms produce different bands?
Isoforms · Possibly. Five isoforms are listed. Isoform 3 lacks residues 162–381; isoform 5 has a replacement at 163–185 and lacks 186–381; isoform 2 has a replacement at 247–254 and lacks 255–381. Isoform 6 replaces 315–381. These sequence differences do not establish which bands will be visible.
Residues 1–161 precede every listed isoform change, making that region a candidate for detecting all five isoforms. Check the antibody’s documented epitope and validate its reactivity; the sequence features alone do not establish antibody performance.
Decide whether the measurement represents total ELMOD3 or a particular isoform. An antibody recognizing a shared region may detect several isoforms; quantify resolved bands separately if isoform differences matter. Do not assign an isoform from band position alone.
Residues 1–161 precede every listed isoform change, making that region a candidate for detecting all five isoforms. Check the antibody’s documented epitope and validate its reactivity; the sequence features alone do not establish antibody performance.
Decide whether the measurement represents total ELMOD3 or a particular isoform. An antibody recognizing a shared region may detect several isoforms; quantify resolved bands separately if isoform differences matter. Do not assign an isoform from band position alone.
Do annotated modifications explain an unexpected ELMOD3 band?
PTM · No modified residues or glycosylation sites are listed in the supplied features. They therefore provide no specific modification or site to test as an explanation. An unexpected band still needs experimental identification.
Does this guide establish induction of ELMOD3?
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 ELMOD3?
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 A12704 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 ELMOD3 be quantified?
Quantitation · Standard workflow guidance: quantify only a validated target band within the linear exposure range. Use consistent sample preparation and loading, retain biological replicates, and avoid interpreting saturation or loading differences as regulation.
How should a band compare with ELMOD3’s predicted mass?
Interpretation · The supplied predicted mass is 43 kDa for the canonical 381-residue sequence. No empirical band position is available, so a migration difference cannot be established from these features.
The listed locations include stereocilia, kinocilia, the cytoplasmic cytoskeleton, and the cuticular plate of auditory hair cells. Consider those structures when selecting a sample or interpreting a fraction, while confirming ELMOD3 recovery experimentally.
Compare its behavior with the listed splice variants and check whether the antibody epitope is retained in each candidate isoform. The predicted canonical mass is 43 kDa, but no observed band size is supplied. Sequence variation alone cannot identify an additional band.
The listed locations include stereocilia, kinocilia, the cytoplasmic cytoskeleton, and the cuticular plate of auditory hair cells. Consider those structures when selecting a sample or interpreting a fraction, while confirming ELMOD3 recovery experimentally.
Compare its behavior with the listed splice variants and check whether the antibody epitope is retained in each candidate isoform. The predicted canonical mass is 43 kDa, but no observed band size is supplied. Sequence variation alone cannot identify an additional band.
Boster reagents
ELMOD3 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-ELMOD3 Antibody
Cat # A12704
The catalog reports one anti-ELMOD3 antibody, A12704, with reported Human, Mouse, and Rat reactivity. Its Western blot image uses extracts from various cell lines at 25 µg per lane and a 1:1000 primary dilution; the caption does not identify the cell lines.
Which to pick: A12704 is the only listed option and has a Western blot image. Check its reported reactivity against your sample; the image caption does not establish which species or cell lines were tested.
Source: BosterBio ELMOD3 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q96FG2.
- Human Protein Atlas. ELMOD3 tissue expression.
- PMC10014252 — target-verified WB comparison