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- Table of Contents
KRT13 / Keratin, type I cytoskeletal 13 · Western blot design guide
Design a Western Blot for KRT13
Source-linked KRT13 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-KRT13 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
KRT13 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 | ~49.6 kDa | |
| Observed band | ~50 kDa | |
| Gel | 10% (catalog M04299) | |
| Positive control | Cervix (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 + Methylated | |
| Caveat | Modification-state controls | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 3 isoform(s) |
Section 1
Source-Linked KRT13 Western Blot Protocol Options
The M04299 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human A431, human Hacat, mouse skin, mouse lung (catalog M04299) |
| Gel % | 10% (catalog M04299) |
| Load | 30 ug; reducing conditions (catalog M04299) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog M04299) |
| Membrane | nitrocellulose membrane (catalog M04299) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog M04299) |
| Primary antibody | M04299 · 1:50000 (catalog M04299) |
| Primary incubation | overnight at 4°C (catalog M04299) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog M04299) |
| Secondary incubation | 1.5 hour at RT (catalog M04299) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog M04299) |
| Detection | ECL (catalog M04299) |
Section 2
What Is the Expected KRT13 Western Blot Band Size?
KRT13 is predicted at 49.6 kDa and observed at ~50 kDa; the cause of the small difference is not established.
| Band at ~50 kDa | empirically observed KRT13 band, close to its 49.6 kDa predicted mass |
| Single sharp band near 50 kDa | consistent with KRT13; its three isoforms need not resolve separately |
| Additional bands at different positions | could reflect isoforms 1, 2, or 3 if they differ in migration; confirm their identity |
| Slight mobility variation near 50 kDa | could reflect Ser427 phosphorylation, though a visible shift is unproven |
💡Expected KRT13 appearanceKRT13 has a predicted mass of 49.6 kDa and an observed band at ~50 kDa; confirm band identity with appropriate controls, since the supplied features do not establish a cause for any mobility difference.
| Predicted KRT13 mass of 49.6 kDa | provides the size reference; the observed band is ~50 kDa |
| Isoform 1 | may differ in size from other isoforms; its separate migration is unestablished |
| Isoform 2 | may differ in size from other isoforms; its separate migration is unestablished |
| Isoform 3 | may differ in size from other isoforms; its separate migration is unestablished |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | KRT13 may remain in an insoluble intermediate filament fraction | check the insoluble fraction and verify extraction with a positive control |
| Band higher than expected | a KRT13 specific mobility shift is not established by the supplied features | confirm identity with a second antibody and compare with a positive control |
| Band lower than expected | an alternative isoform or protein breakdown is possible | compare fresh samples and confirm identity with another antibody |
| Multiple bands | isoforms 1, 2, and 3 are listed, but separate bands are unproven | compare isoform aware controls and confirm each band independently |
| Weak or no signal | incomplete recovery of intermediate filament KRT13 is possible | check extraction and loading against a KRT13 positive control |
| Fragments below expected size | sample degradation is possible; no cleavage product is specified | prepare fresh samples with protease inhibitors and confirm fragment identity |
Experiment evidence
Western blot images showing KRT13 band
M04299 · KRT13
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for KRT13 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for KRT13 in Western blot, you can use cervix tissue, which HPA scores as High.
Positive control: Cervix (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 cervix and no detection in adipose tissue, so tissue controls are feasible.
HPA tissue expression evidence for KRT13
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 |
|---|---|---|---|---|
| Cervix | squamous epithelial cells | High | Protein (IHC) | HPA → |
| Esophagus | squamous epithelial cells | High | Protein (IHC) | HPA → |
| Oral mucosa | squamous epithelial cells | High | Protein (IHC) | HPA → |
| Skin | cells in basal layer | High | Protein (IHC) | HPA → |
| Tonsil | squamous epithelial 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 → |
| Adrenal gland | glandular cells | Not detected | Protein (IHC) | HPA → |
| Appendix | glandular cells | Not detected | Protein (IHC) | HPA → |
| Bone marrow | hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Breast | adipocytes | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced KRT13 Western Blot Tips
Deeper troubleshooting and optimisation questions for KRT13, answered from its protein features.
How should KRT13 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 KRT13 isoforms produce different bands?
Isoforms · UniProt lists three isoforms. Relative to the 458-residue canonical sequence, isoform 2 lacks residues 62–73 and replaces residues 351–458; isoform 3 replaces residues 416–458. These sequence changes could affect apparent size, but the features do not establish where each isoform runs.
Check the antibody epitope against the alternative sequences. Canonical residues 351–458 are replaced in isoform 2, and residues 416–458 are replaced in isoform 3. An antibody recognizing either changed region may detect the isoforms differently.
Check the antibody epitope against the alternative sequences. Canonical residues 351–458 are replaced in isoform 2, and residues 416–458 are replaced in isoform 3. An antibody recognizing either changed region may detect the isoforms differently.
How should phosphorylation at KRT13 Ser427 be interpreted?
PTM · UniProt lists phosphoserine at canonical residue 427. That residue lies within sequence replaced in isoforms 2 and 3, so do not assign the canonical Ser427 site to either alternative sequence without checking its numbering. The feature alone does not predict a visible band shift.
Does this guide establish induction of KRT13?
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 KRT13?
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 M04299 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 can KRT13 bands be quantified consistently?
Quantitation · Quantify the same defined band near 50 kDa across samples, since that is the reported apparent band and is close to the 49.6 kDa predicted mass. If additional bands appear, assess them separately; the three listed isoforms and modified residues do not by themselves identify those bands.
Does the ~50 kDa band indicate a KRT13 mass shift?
Interpretation · The observed band near 50 kDa agrees with the predicted 49.6 kDa mass. The listed modifications alone do not establish a visible shift or explain any small difference between apparent and calculated mass.
UniProt lists omega-N-methylarginine at canonical residues 27 and 35. Both positions precede the listed isoform sequence changes. Their presence does not establish that methylation produces a resolvable band shift.
Consider the listed isoform sequence changes and whether the antibody recognizes each sequence. KRT13 also forms heterotetramers with type II keratins and has listed methylation and phosphorylation sites. None of these features alone identifies an unexpected band; avoid assigning it to a specific isoform or modification from position alone.
UniProt lists omega-N-methylarginine at canonical residues 27 and 35. Both positions precede the listed isoform sequence changes. Their presence does not establish that methylation produces a resolvable band shift.
Consider the listed isoform sequence changes and whether the antibody recognizes each sequence. KRT13 also forms heterotetramers with type II keratins and has listed methylation and phosphorylation sites. None of these features alone identifies an unexpected band; avoid assigning it to a specific isoform or modification from position alone.
Boster reagents
KRT13 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 13 Rabbit Monoclonal Antibody
Cat # M04299
The catalog reports one anti-KRT13 Western blot antibody, M04299, with reported human and mouse reactivity. Its WB image shows an approximately 50 kDa band in human A431 and HaCaT cell lysates and mouse skin and lung lysates; evidence for other samples is not supplied.
Which to pick: M04299 is the only listed option. Its WB image includes human cell lysates and mouse tissue lysates, making those the documented starting contexts. The caption reports 30 µg per lane under reducing conditions and a 1:50,000 primary antibody dilution.
Source: BosterBio KRT13 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P13646.
- Human Protein Atlas. KRT13 tissue expression.
- PMC7864948 — target-verified WB comparison
- PMC7206110 — target-verified WB comparison
- PMC8916577 — target-verified WB comparison