This website uses cookies to ensure you get the best experience on our website.
- Table of Contents
CYP26B1 / Cytochrome P450 26B1 · Western blot design guide
Design a Western Blot for CYP26B1
Source-linked CYP26B1 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-CYP26B1 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
CYP26B1 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 | ~57.5 kDa | |
| Observed band | ~58 kDa | |
| Gel | 10% (catalog A02691-2) | |
| 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 | — | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 3 isoform(s) |
Section 1
Source-Linked CYP26B1 Western Blot Protocol Options
The A02691-2 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human U2OS, human Hacat, human A549, rat brain, mouse brain (catalog A02691-2) |
| Gel % | 10% (catalog A02691-2) |
| Load | 30 ug; reducing conditions (catalog A02691-2) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog A02691-2) |
| Membrane | nitrocellulose membrane (catalog A02691-2) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog A02691-2) |
| Primary antibody | A02691-2 · 0.5 μg/mL (catalog A02691-2) |
| Primary incubation | overnight at 4°C (catalog A02691-2) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog A02691-2) |
| Secondary incubation | 1.5 hour at RT (catalog A02691-2) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog A02691-2) |
| Detection | ECL (catalog A02691-2) |
Section 2
What Is the Expected CYP26B1 Western Blot Band Size?
CYP26B1 is predicted at 57.5 kDa and observed at ~58 kDa; the cause of the small difference is not established.
| Band at ~58 kDa | Empirical CYP26B1 band in whole-cell and tissue lysates |
| Band near 57.5 kDa | Consistent with the predicted protein mass |
| Several bands at different positions | Could reflect isoforms 1, 2, and 3, but their migration is uncharacterized |
| Weak band in a membrane-poor fraction | Consistent with CYP26B1 localization to ER and microsome membranes |
💡Expected CYP26B1 appearanceCYP26B1 has a predicted mass of 57.5 kDa and an empirical band at ~58 kDa; confirm band identity with antibody specificity controls, since the cause of any migration difference is unestablished.
| Predicted protein mass | 57.5 kDa predicted; the empirical band is ~58 kDa |
| Splice isoform 1 | Its individual mass and migration are not supplied |
| Splice isoform 2 | May differ in size from other isoforms; its mass is not supplied |
| Splice isoform 3 | May differ in size from other isoforms; its mass is not supplied |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | ER and microsome membrane protein may be poorly recovered | Check a membrane-enriched fraction and confirm extraction |
| Band higher than expected | An alternate isoform or unrelated antibody signal is possible | Compare with an isoform-resolved sample and verify antibody specificity |
| Band lower than expected | An alternate isoform or unrelated antibody signal is possible | Use an independent antibody or CYP26B1 depletion control |
| Multiple bands | Isoforms 1, 2, and 3 are listed, but their band positions are unknown | Test isoform-specific expression and band identity |
| Weak or no signal | Membrane extraction may limit CYP26B1 recovery | Optimize membrane solubilization and check a microsome-enriched sample |
Experiment evidence
Western blot images showing CYP26B1 band
A02691-2 · CYP26B1
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for CYP26B1 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for CYP26B1 in Western blot, you can use adrenal gland tissue, an HPA positive sample.
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: CYP26B1 is ER/microsome-associated, so membrane-enriched lysate may improve detection.
HPA tissue expression evidence for CYP26B1
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 → |
| Caudate | neuronal cells | Medium | Protein (IHC) | HPA → |
| Cerebellum | Purkinje cells | Medium | Protein (IHC) | HPA → |
| Cerebral cortex | neuronal cells | Medium | Protein (IHC) | HPA → |
| Cervix | glandular cells | Medium | 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 → |
| Endometrium | cells in endometrial stroma | Not detected | Protein (IHC) | HPA → |
| Esophagus | squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
| Fallopian tube | glandular cells | Not detected | Protein (IHC) | HPA → |
| Heart muscle | cardiomyocytes | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced CYP26B1 Western Blot Tips
Deeper troubleshooting and optimisation questions for CYP26B1, answered from its protein features.
How should CYP26B1 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 CYP26B1 isoforms produce different bands?
Isoforms · UniProt lists three isoforms. Relative to the canonical sequence, isoform 2 lacks residues 69–143, while isoform 3 replaces residues 1–67. These are UniProt canonical-sequence coordinates. Different migration is plausible, but the features do not establish where each isoform runs on a blot.
Check the antibody epitope against the isoform sequences. An epitope within canonical residues 69–143 would be absent from isoform 2; one within canonical residues 1–67 may differ in isoform 3. These coordinates use UniProt canonical numbering and may differ from antibody documentation.
Check the antibody epitope against the isoform sequences. An epitope within canonical residues 69–143 would be absent from isoform 2; one within canonical residues 1–67 may differ in isoform 3. These coordinates use UniProt canonical numbering and may differ from antibody documentation.
Is a modification responsible for a shifted CYP26B1 band?
PTM · The supplied features list no modified residues, glycosylation sites, or disulfides. They therefore provide no specific modification to assign to a shifted band. A sequence feature alone would not demonstrate a visible shift; investigate band identity before attributing one to a modification.
Does this guide establish induction of CYP26B1?
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 CYP26B1 Western blot?
Transfer · CYP26B1 is annotated at endoplasmic reticulum and microsome membranes, with a predicted mass of 57.5 kDa. Include the membrane-containing fraction during preparation and verify transfer around 58 kDa with total-protein staining. The supplied features do not specify a particular transfer method.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A02691-2 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 CYP26B1 bands be quantified?
Quantitation · Quantify a defined band consistently across samples and check whether the antibody detects multiple isoforms. Normalize to an appropriate loading measure, especially when comparing membrane-containing fractions. The ~58 kDa position is consistent with the predicted mass but does not, by itself, establish which isoform the band represents.
Does the ~58 kDa band match CYP26B1’s predicted mass?
Interpretation · Yes. The observed ~58 kDa band is close to the predicted 57.5 kDa. This agreement supports the assignment but does not establish band identity; confirm it with an appropriate specificity control.
First consider the three annotated isoforms and check the antibody epitope against their sequences. Isoform 2 lacks canonical residues 69–143; isoform 3 replaces canonical residues 1–67. UniProt lists no specific modification that explains another band. Use a specificity control before assigning any unexpected band to CYP26B1.
First consider the three annotated isoforms and check the antibody epitope against their sequences. Isoform 2 lacks canonical residues 69–143; isoform 3 replaces canonical residues 1–67. UniProt lists no specific modification that explains another band. Use a specificity control before assigning any unexpected band to CYP26B1.
Boster reagents
CYP26B1 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-CYP26B1 Antibody Picoband®
Cat # A02691-2
The catalog reports one anti-CYP26B1 antibody for Western blot, A02691-2, with reported human, mouse and rat reactivity. Its product image shows an approximately 58 kDa band; no publication evidence was supplied.
Which to pick: A02691-2 is the only listed option. Its WB image uses human U2OS, HaCaT and A549 cell lysates and rat and mouse brain lysates, with 30 µg per lane and antibody at 0.5 µg/mL. These examples do not establish performance in every sample.
Source: BosterBio CYP26B1 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q9NR63.
- Human Protein Atlas. CYP26B1 tissue expression.
- PMC13579854 — target-verified WB comparison
- PMC11788749 — target-verified WB comparison
- PMC3362586 — target-verified WB comparison