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- Table of Contents
FADS2 / Acyl-CoA 6-desaturase · Western blot design guide
Design a Western Blot for FADS2
Real validated FADS2 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-FADS2 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
FADS2 Western Blot Experimental Design Guide
Expected bands, validated protocols, controls and antibodies — the at-a-glance facts below, then the full design guide.
Must know before running
| Expected band | ~52.3 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 | — | |
| Caveat | Membrane extraction controls | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 4 isoform(s) |
Section 1
Real Curated FADS2 Western Blot Protocols
The A01220-1 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | Target-positive lysate and matched negative control (standard starting point) |
| 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 | A01220-1; 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 FADS2 Western Blot Band Size?
FADS2 has a predicted mass of 52.3 kDa; four isoforms could affect band patterns, but distinct migration has not been demonstrated here.
| Band near 52.3 kDa | consistent with the predicted FADS2 mass; confirm identity with controls |
| Several bands at different positions | could include isoforms 1, 2, 3, or 4; their migration is unknown |
| Little or no band in a soluble fraction | FADS2 is an endoplasmic reticulum membrane protein |
| Band in a membrane-enriched fraction | consistent with FADS2 membrane localization; identity still requires confirmation |
💡Expected FADS2 appearanceUniProt predicts 52.3 kDa for FADS2; no empirical band size or isoform migration is supplied, so confirm any candidate band with ordinary identity controls.
| Predicted FADS2 mass | sets a 52.3 kDa reference, not a measured migration position |
| Isoform 1 | its separate apparent size is not supplied |
| Isoform 2 | its size relative to other isoforms is unknown |
| Isoform 3 | its size relative to other isoforms is unknown |
| Isoform 4 | its size relative to other isoforms is unknown |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | ER membrane FADS2 may be poorly recovered during extraction | check membrane recovery and test a membrane-enriched fraction |
| Band higher than expected | the identity and migration of this band are unestablished | compare with a molecular-weight marker and verify identity with an independent antibody or FADS2 depletion |
| Band lower than expected | an alternative isoform is possible, but its mass is unknown | verify identity with FADS2 depletion and compare antibody epitopes |
| Multiple bands | four annotated isoforms could contribute, but distinct migration is unproven | check which bands respond to FADS2 depletion |
| Weak or no signal | membrane-associated FADS2 may be underrepresented in the prepared sample | check extraction efficiency and compare a membrane-enriched sample |
Experiment evidence
Western blot images showing FADS2 band
A01220-1 · FADS2
A01220 · FADS2
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for FADS2 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for FADS2 in Western blot, you can use adrenal gland tissue.
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: As an ER membrane protein, FADS2 may be easier to detect in membrane-enriched lysate.
HPA tissue expression evidence for FADS2
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 | High | Protein (IHC) | HPA → |
| Appendix | glandular cells | High | Protein (IHC) | HPA → |
| Cerebellum | Purkinje cells | High | Protein (IHC) | HPA → |
| Duodenum | glandular cells | High | Protein (IHC) | HPA → |
| Epididymis | 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 → |
| Lymph node | germinal center cells | Not detected | Protein (IHC) | HPA → |
| Ovary | ovarian stroma cells | Not detected | Protein (IHC) | HPA → |
| Parathyroid gland | glandular cells | Not detected | Protein (IHC) | HPA → |
| Spleen | cells in red pulp | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced FADS2 Western Blot Tips
Deeper troubleshooting and optimisation questions for FADS2, answered from its protein features.
How should FADS2 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 FADS2 isoforms produce different bands?
Isoforms · UniProt lists four isoforms. Isoforms 2 and 4 replace canonical residues 1–69 with different N-terminal sequences; isoform 3 replaces residues 386–444 with Q. These changes could affect size or antibody recognition, but the features do not show which isoforms are present in your sample or where they migrate.
For coverage of all listed isoforms, choose an epitope outside canonical residues 1–69 and 386–444, then verify the antibody's stated specificity. An epitope within either changed region may distinguish isoforms or miss some of them. These coordinates use the supplied UniProt canonical sequence numbering.
For coverage of all listed isoforms, choose an epitope outside canonical residues 1–69 and 386–444, then verify the antibody's stated specificity. An epitope within either changed region may distinguish isoforms or miss some of them. These coordinates use the supplied UniProt canonical sequence numbering.
Do annotated modifications prove a band shift?
PTM · The linked UniProt record describes protein features. A modification annotation alone does not demonstrate a visible shift; retain any condition or experimental qualifier attached to it.
Does this guide establish induction of FADS2?
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 FADS2 Western blot?
Transfer · FADS2 is an ER multi-pass membrane protein. Check that it is extracted from the membrane fraction and transfers efficiently by inspecting the post-transfer gel and membrane. The supplied features do not specify a transfer membrane, buffer, or time, so optimize those against recovery of the approximately 52.3 kDa canonical protein.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A01220-1 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 FADS2 bands be quantified?
Quantitation · Quantify bands within the assay's linear signal range and apply the same band-inclusion rule across samples. Because four isoforms are listed, decide whether the antibody and chosen band represent one isoform or a combined FADS2 signal. Confirm that the extraction method recovers this ER membrane protein consistently.
Where should FADS2 appear relative to its predicted mass?
Interpretation · The canonical 444-residue sequence has a predicted mass of 52.3 kDa. No observed band position is supplied. Its ER membrane location and alternative sequences warrant checking band identity, but the supplied features do not establish a visible shift from 52.3 kDa.
Check antibody epitope coverage and compare the bands with the listed isoform changes before assigning identities. The supplied features list no glycosylation or modified-residue sites, and no observed band position is available. Isoform features alone cannot establish the cause of an extra band or a mass difference.
Check antibody epitope coverage and compare the bands with the listed isoform changes before assigning identities. The supplied features list no glycosylation or modified-residue sites, and no observed band position is available. Isoform features alone cannot establish the cause of an extra band or a mass difference.
Boster reagents
FADS2 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-FADS2 Antibody (Center)
Cat # A01220-1
Real WB data
Anti-Fatty acid desaturase 2 FADS2 Antibody
Cat # A01220
Both listed anti-FADS2 antibodies have Western blot images. A01220-1 was shown with CEM cell lysate (35 µg/lane); A01220 was shown with A549, HeLa, mouse liver, and mouse brain lysates. These examples document tested samples, not broad validation.
Which to pick: For human CEM samples, consider A01220-1. For the listed human cell lines or mouse tissues, consider A01220. A01220 also lists rat reactivity, but no rat Western blot example is supplied.
Source: BosterBio FADS2 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry O95864.
- Human Protein Atlas. FADS2 tissue expression.
- PMC12170995 — target-verified WB comparison
- PMC12391508 — target-verified WB comparison
- PMC9065188 — target-verified WB comparison