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- Table of Contents
KRT15 / Keratin, type I cytoskeletal 15 · Western blot design guide
Design a Western Blot for KRT15
Source-linked KRT15 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-KRT15 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
KRT15 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 | ~49.2 kDa | |
| Observed band | ~49 kDa | |
| Gel | 10% (catalog M06791-1) | |
| Positive control | Breast (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 + Ubl conjugation | |
| Caveat | Phosphorylation-state controls | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 2 isoform(s) |
Section 1
Source-Linked KRT15 Western Blot Protocol Options
The M06791-1 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human Hacat, human A431, human PC-3 (catalog M06791-1) |
| Gel % | 10% (catalog M06791-1) |
| Load | 30 ug; reducing conditions (catalog M06791-1) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog M06791-1) |
| Membrane | nitrocellulose membrane (catalog M06791-1) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog M06791-1) |
| Primary antibody | M06791-1 · 1:1000 (catalog M06791-1) |
| Primary incubation | overnight at 4°C (catalog M06791-1) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog M06791-1) |
| Secondary incubation | 1.5 hour at RT (catalog M06791-1) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog M06791-1) |
| Detection | ECL (catalog M06791-1) |
Section 2
What Is the Expected KRT15 Western Blot Band Size?
KRT15 is predicted at 49.2 kDa and observed at ~49 kDa; its isoforms and phosphorylation sites have no demonstrated migration effect here.
| Band at ~49 kDa | Matches the empirical KRT15 band and the 49.2 kDa predicted mass. |
| Two bands near 49 kDa | Isoforms 1 and 2 are possible contributors, but distinct migration is unproven. |
| Band above 49 kDa | Its identity is uncertain; KRT15 phosphorylation sites do not establish a visible shift. |
| Weak or absent lysate band | KRT15 is an intermediate filament protein that may be poorly recovered during extraction. |
💡Expected KRT15 appearanceExpect a KRT15 band at the empirical ~49 kDa, close to its 49.2 kDa predicted mass; confirm identity with an appropriate positive control or knockdown.
| UniProt predicted mass | Places the predicted KRT15 protein at 49.2 kDa, consistent with the observed ~49 kDa band. |
| Splice isoform 1 | May differ in size from isoform 2; its individual mass and migration are not supplied. |
| Splice isoform 2 | May differ in size from isoform 1; a separate visible band is not established. |
| Phosphoserine sites at residues 17, 34, 36, 48, and 56 | Could affect apparent migration, but no shift is demonstrated. |
| Phosphothreonine sites at residues 301 and 323 | Could affect apparent migration, but no shift is demonstrated. |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Intermediate filament KRT15 may be poorly recovered during extraction. | Check the insoluble fraction and run a KRT15 positive control. |
| Band higher than expected | The cause is unestablished; phosphorylation alone does not verify the band's identity. | Check reduction and compare with a positive control or KRT15 knockdown. |
| Band lower than expected | The lower band is not explained by a supplied isoform mass. | Check band identity with KRT15 knockdown and assess sample degradation. |
| Multiple bands | Isoforms 1 and 2 exist, but separate migration is unproven. | Use KRT15 knockdown to identify specific bands. |
| Weak or no signal | Intermediate filament recovery may be low. | Assess the insoluble fraction and confirm the assay with a positive control. |
| Fragments below expected size | Sample proteolysis is possible; no KRT15 fragment mass is supplied. | Prepare fresh lysate with protease inhibitors and verify bands by KRT15 knockdown. |
Experiment evidence
Western blot images showing KRT15 band
M06791-1 · KRT15
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for KRT15 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for KRT15 in Western blot, you can use breast tissue, which has high HPA expression.
Positive control: Breast (IHC candidate; verify WB)
Negative control: Adipose tissue (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain such as Ponceau alongside the samples.
⚠️Feasibility: HPA reports high expression in breast and no detection in adipose tissue, supporting a practical tissue control pair.
HPA tissue expression evidence for KRT15
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 |
|---|---|---|---|---|
| Breast | glandular cells | High | Protein (IHC) | HPA → |
| Bronchus | respiratory epithelial cells | High | Protein (IHC) | HPA → |
| Esophagus | squamous epithelial cells | High | Protein (IHC) | HPA → |
| Hair | cells in external root sheath | High | Protein (IHC) | HPA → |
| Skin | secretory 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 → |
| Adrenal gland | 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 → |
Section 3
Advanced KRT15 Western Blot Tips
Deeper troubleshooting and optimisation questions for KRT15, answered from its protein features.
How should KRT15 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 KRT15 isoforms produce different bands?
Isoforms · UniProt lists isoforms 1 and 2. Isoform 2 replaces residues 3–29 and lacks residues 30–194 and 417–456 of the canonical sequence, so a smaller band is plausible. The features alone do not establish its apparent position on a blot.
Check the antibody epitope against both isoforms. An epitope within canonical residues 30–194 or 417–456 is absent from isoform 2; residues 3–29 are replaced. Recognition cannot be assumed from the isoform names alone.
No. Canonical residue 17 lies in the segment replaced in isoform 2, while residues 34, 36, 48 and 56 lie in a segment missing from it. Canonical residues 301 and 323 are retained, but their isoform 2 coordinates differ. State the sequence convention when comparing site-specific results.
Check the antibody epitope against both isoforms. An epitope within canonical residues 30–194 or 417–456 is absent from isoform 2; residues 3–29 are replaced. Recognition cannot be assumed from the isoform names alone.
No. Canonical residue 17 lies in the segment replaced in isoform 2, while residues 34, 36, 48 and 56 lie in a segment missing from it. Canonical residues 301 and 323 are retained, but their isoform 2 coordinates differ. State the sequence convention when comparing site-specific results.
Which KRT15 phosphorylation sites are listed?
PTM · UniProt lists phosphoserine at positions 17, 34, 36, 48 and 56, and phosphothreonine at 301 and 323. These are canonical UniProt coordinates, which may differ from antibody or paper numbering. Their presence does not establish a visible band shift.
Does this guide establish induction of KRT15?
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 KRT15?
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 M06791-1 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 KRT15 bands be quantified?
Quantitation · Define which detected band or bands are included and apply that choice consistently across samples. The two listed isoforms differ substantially in sequence, so confirm that the antibody recognizes the forms being compared before treating band intensity as total KRT15.
Does the observed KRT15 band match its predicted mass?
Interpretation · The reported band near 49 kDa agrees with the 49.2 kDa predicted mass. The listed features do not establish a measurable shift from that value.
Consider the listed alternative isoform and phosphorylation sites, then check antibody recognition before assigning an identity. KRT15 forms keratin complexes, including a heterodimer with KRT14, but these features alone do not identify an unexpected band or demonstrate a phosphorylation-dependent shift.
Consider the listed alternative isoform and phosphorylation sites, then check antibody recognition before assigning an identity. KRT15 forms keratin complexes, including a heterodimer with KRT14, but these features alone do not identify an unexpected band or demonstrate a phosphorylation-dependent shift.
Boster reagents
KRT15 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-Cytokeratin 15 Rabbit Monoclonal Antibody
Cat # M06791-1
M06791-1 is a rabbit monoclonal anti-KRT15 antibody listed for human, mouse, and rat. Its Western blot image shows an approximately 49 kDa band in human HaCaT, A431, and PC-3 whole cell lysates; the supplied image does not test mouse or rat samples.
Which to pick: M06791-1 is the only listed option. Choose it when its stated reactivity fits your sample; its supplied Western blot image documents human cell lysates at a 1:1000 primary antibody dilution.
Source: BosterBio KRT15 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P19012.
- Human Protein Atlas. KRT15 tissue expression.
- PMC9302346 — target-verified WB comparison
- PMC7466560 — target-verified WB comparison
- PMC10675954 — target-verified WB comparison