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ELOVL5 / Very long chain fatty acid elongase 5 · Western blot design guide
Design a Western Blot for ELOVL5
Source-linked ELOVL5 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-ELOVL5 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
ELOVL5 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 | ~35.3 kDa | |
| Gel | 12–15% (standard starting point) | |
| Positive control | Adrenal gland (IHC candidate; verify WB) +4 more | |
| Negative control | Adipose tissue (IHC candidate; verify WB) |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Phosphorylated + Acetylated | |
| Caveat | Modification-state controls | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 3 isoform(s) |
Section 1
Source-Linked ELOVL5 Western Blot Protocol Options
The M03615 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | (1) HeLa cell lysate; (2) RAW 264.7 cell lysate (catalog M03615) |
| 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 | 5% milk or 5% BSA in TBST (standard starting point) |
| Primary antibody | M03615; use the WB datasheet starting dilution (standard starting point) |
| Primary incubation | Overnight at 4 °C (standard starting point) |
| Secondary antibody | Species-matched HRP conjugate at validated dilution (standard starting point) |
| Secondary incubation | 1 h at room temperature (standard starting point) |
| Wash | 3 × 5 min in TBST (standard starting point) |
| Detection | ECL; bracket exposures to avoid saturation (standard starting point) |
Section 2
What Is the Expected ELOVL5 Western Blot Band Size?
ELOVL5 is predicted at 35.3 kDa; annotated isoforms and membrane localization may affect blot appearance, but no empirical migration or distinct isoform bands are established.
| Band near 35.3 kDa | consistent with the predicted ELOVL5 size; confirm identity with controls |
| Additional bands at different positions | could reflect isoforms 1, 2, or 3, but their migration is unknown |
| Weak or absent band in soluble lysate | ELOVL5 is a multipass endoplasmic reticulum membrane protein |
| Stronger band in a membrane-enriched fraction | consistent with ELOVL5 membrane localization |
💡Expected ELOVL5 appearanceUniProt predicts 35.3 kDa, but no empirical band size is supplied; isoforms 1, 2, and 3 may affect migration without a demonstrated band pattern, so confirm band identity with controls.
| UniProt predicted mass | places the reference size at 35.3 kDa |
| Isoform 1 | may differ in size from other isoforms; its relative size is unknown |
| Isoform 2 | may differ in size from other isoforms; its relative size is unknown |
| Isoform 3 | may differ in size from other isoforms; its relative size is unknown |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | ELOVL5 may be poorly recovered from the membrane fraction | check membrane enrichment and detergent extraction |
| Band higher than expected | an isoform or an unrelated band could migrate higher | compare fractions and verify identity with an independent antibody or knockdown |
| Band lower than expected | an isoform or a fragment could migrate lower | check sample integrity and verify identity with knockdown |
| Multiple bands | isoforms 1, 2, and 3 are annotated, but distinct bands are unproven | verify each band with isoform-aware controls or knockdown |
| Weak or no signal | membrane protein recovery may be low | assess extraction efficiency and enrich membranes |
Experiment evidence
Western blot images showing ELOVL5 band
M03615 · ELOVL5
A30674 · ELOVL5
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for ELOVL5 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for ELOVL5 in Western blot, you can use adrenal gland tissue, which HPA rates High.
Positive control: Adrenal gland (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: Because ELOVL5 is an ER membrane protein, membrane-enriched lysates may improve detection.
HPA tissue expression evidence for ELOVL5
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 | cells in zona fasciculata | High | Protein (IHC) | HPA → |
| Appendix | glandular cells | High | Protein (IHC) | HPA → |
| Breast | glandular cells | High | Protein (IHC) | HPA → |
| Prostate | glandular cells | High | Protein (IHC) | HPA → |
| Seminal vesicle | glandular cells | 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 → |
| Bone marrow | hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Heart muscle | cardiomyocytes | Not detected | Protein (IHC) | HPA → |
| Lymph node | germinal center cells | Not detected | Protein (IHC) | HPA → |
| Ovary | follicle cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced ELOVL5 Western Blot Tips
Deeper troubleshooting and optimisation questions for ELOVL5, answered from its protein features.
What should be checked when unexpected ELOVL5 bands appear?
Band shift · Check antibody epitope coverage against all three isoforms, particularly the extensive sequence loss in isoform 3. Compare candidate bands with a specific control. UniProt lists canonical phosphoserine 285 and N-acetylmethionine 1, but their presence alone cannot explain a band shift or establish the identity of an unexpected band.
How could ELOVL5 isoforms affect band interpretation?
Isoforms · UniProt lists three isoforms. Isoform 2 has an alternative sequence at canonical residue 82; isoform 3 replaces residues 20–88 and lacks residues 89–299. Check whether the antibody epitope is present in each isoform before assigning bands. These sequence differences do not establish which isoforms are expressed in your sample.
Which ELOVL5 modifications matter for phospho-specific detection?
PTM · UniProt lists N-acetylmethionine at position 1 and phosphoserine at position 285, using canonical sequence numbering. Isoform 3 lacks the canonical region containing position 285; isoform 2 changes sequence at position 82, so downstream numbering may differ. A phospho-specific result needs site-specific validation; the listed modifications alone do not predict a visible shift.
For an antibody targeting the listed phosphoserine at canonical position 285, compare blocking conditions using the same samples and exposure settings. Choose the condition that preserves specific signal while reducing background, and validate site specificity before interpreting signal differences as changes in phosphorylation.
For an antibody targeting the listed phosphoserine at canonical position 285, compare blocking conditions using the same samples and exposure settings. Choose the condition that preserves specific signal while reducing background, and validate site specificity before interpreting signal differences as changes in phosphorylation.
Does this guide establish induction of ELOVL5?
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 ELOVL5 Western blot?
Transfer · ELOVL5 is an endoplasmic reticulum, multipass membrane protein. Check that sample preparation solubilizes it and that transfer moves the candidate band onto the membrane. Compare transfer conditions empirically with a suitable control; the supplied features do not identify one method as best.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the M03615 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 ELOVL5 Western blot signal be quantified?
Quantitation · Quantify a validated ELOVL5 band within the assay’s linear signal range. Because UniProt lists three isoforms with different sequences, confirm which forms the antibody recognizes before combining bands. For phosphoserine 285 measurements, report the phospho signal relative to a validated total ELOVL5 measurement.
Should ELOVL5 migrate at exactly 35.3 kDa?
Interpretation · 35.3 kDa is the predicted mass of canonical ELOVL5. Its multiple membrane spans and listed modifications do not establish where it will migrate on a Western blot. No observed band mass is supplied, so compare candidate bands with appropriate controls rather than assigning identity by size alone.
Boster reagents
ELOVL5 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-ELOVL5 Rabbit Monoclonal Antibody
Cat # M03615
Real WB data
Anti-ELOVL5 Antibody
Cat # A30674
The catalog reports two anti-ELOVL5 antibodies, M03615 and A30674, with reported Human, Mouse, and Rat reactivity. Both have Western blot images. The captions show specific cell lysates; they do not establish performance across every listed species or sample type.
Which to pick: Choose M03615 if the HeLa and RAW 264.7 lysate examples fit your work. Choose A30674 for an MCF-7 lysate example with a peptide-blocked lane. Both list Human, Mouse, and Rat reactivity; match your sample to the reported conditions.
Source: BosterBio ELOVL5 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q9NYP7.
- Human Protein Atlas. ELOVL5 tissue expression.
- PMC6277159 — target-verified WB comparison
- PMC3031740 — target-verified WB comparison