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- Table of Contents
KRT6A / Keratin, type II cytoskeletal 6A · Western blot design guide
Design a Western Blot for KRT6A
Source-linked KRT6A Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-KRT6A 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
KRT6A 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 | ~60 kDa | |
| Observed band | ~60 kDa | |
| Gel | 10% (catalog M30943-1) | |
| 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 | Acetylated | |
| Caveat | — | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 1 isoform(s) |
Section 1
Source-Linked KRT6A Western Blot Protocol Options
The M30943-1 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 (catalog M30943-1) |
| Gel % | 10% (catalog M30943-1) |
| Load | 30 ug; reducing conditions (catalog M30943-1) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog M30943-1) |
| Membrane | nitrocellulose membrane (catalog M30943-1) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog M30943-1) |
| Primary antibody | M30943-1 · 1: 500 (catalog M30943-1) |
| Primary incubation | overnight at 4°C (catalog M30943-1) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog M30943-1) |
| Secondary incubation | 1.5 hour at RT (catalog M30943-1) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog M30943-1) |
| Detection | ECL (catalog M30943-1) |
Section 2
What Is the Expected KRT6A Western Blot Band Size?
KRT6A is predicted at 60 kDa, and antibody QC reports a Cytokeratin 6 band at approximately 60 kDa; confirm KRT6A identity with a specificity control.
| Band near 60 kDa in A431 lysate | Matches the predicted KRT6A mass and the reported Cytokeratin 6 band |
| Band near 60 kDa in HaCaT lysate | Matches the predicted KRT6A mass and the reported Cytokeratin 6 band |
| Bands near 60 kDa in both lysates | Matches the reported Cytokeratin 6 result in both samples |
| Band near 60 kDa under reducing conditions | Consistent with the predicted KRT6A mass; identity still requires a specificity control |
💡Expected KRT6A appearanceKRT6A is predicted at 60 kDa, and antibody QC reports a Cytokeratin 6 band at approximately 60 kDa in reducing whole-cell lysates; confirm KRT6A identity with a specificity control.
| UniProt predicted molecular weight | Places KRT6A near 60 kDa |
| Full-length predicted mass | Corresponds to approximately 60 kDa |
| Predicted mass of 60,045 Da | Corresponds to approximately 60 kDa |
| Predicted mass for the 564-residue sequence | Provides a reference near 60 kDa |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | KRT6A expression or recovery may be low | Check expression and extraction with a positive lysate control |
| Band higher than expected | Band identity or migration is uncertain | Confirm the band with KRT6A depletion or an independent antibody |
| Band lower than expected | Possible degradation or an unrelated band | Use protease inhibitors and confirm identity with KRT6A depletion |
| Multiple bands | Possible recognition of related keratins or unrelated proteins | Compare bands after KRT6A depletion |
| Weak or no signal | Low KRT6A abundance or poor extraction is possible | Check sample recovery and include a positive lysate control |
Experiment evidence
Western blot images showing KRT6A band
M30943-1 · KRT6A
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for KRT6A Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for KRT6A in Western blot, you can use cervix tissue, which HPA scores 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 alongside the samples.
⚠️Feasibility: HPA reports a clear negative tissue, but use a high-expression tissue for the positive control.
HPA tissue expression evidence for KRT6A
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 → |
| Nasopharynx | respiratory epithelial cells | High | Protein (IHC) | HPA → |
| Oral mucosa | squamous epithelial cells | High | Protein (IHC) | HPA → |
| Skin | cells in corneal layer | 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 → |
| 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 KRT6A Western Blot Tips
Deeper troubleshooting and optimisation questions for KRT6A, answered from its protein features.
How should KRT6A 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.
Should KRT6A isoforms produce multiple bands?
Isoforms · The supplied record lists one isoform and no alternative sequence. It does not support assigning multiple bands to KRT6A isoforms.
Which KRT6A modification should be considered when interpreting a band?
PTM · UniProt lists N-acetylalanine at position 2. The sequence begins MA, consistent with that coordinate. State that you are using UniProt numbering if comparing it with antibody or paper numbering.
The listed position 2 acetylation alone does not establish a visible shift. Since the reported band matches the predicted mass, do not attribute another band to acetylation without additional evidence.
The listed position 2 acetylation alone does not establish a visible shift. Since the reported band matches the predicted mass, do not attribute another band to acetylation without additional evidence.
Does this guide establish induction of KRT6A?
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 KRT6A?
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 M30943-1 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.
Which band should be quantified for KRT6A?
Quantitation · Use the reported approximately 60 kDa band as the candidate KRT6A signal. Keep the band definition consistent across samples, and validate any additional band before including it in KRT6A quantitation.
How does the observed KRT6A band compare with its predicted mass?
Interpretation · The reported band is approximately 60 kDa, matching the predicted 60 kDa. Use this as a starting point for identifying KRT6A, but confirm band identity independently.
KRT6A is described as forming a heterodimer with a type I keratin; KRT6 isomers associate with KRT16 and/or KRT17. These interactions provide context, but do not establish that a higher Western blot band is a complex.
Compare them with the reported approximately 60 kDa band. The supplied features do not establish alternative KRT6A isoforms, glycosylation sites, or a specific cause of extra bands, so verify their identity before assigning them to KRT6A.
KRT6A is described as forming a heterodimer with a type I keratin; KRT6 isomers associate with KRT16 and/or KRT17. These interactions provide context, but do not establish that a higher Western blot band is a complex.
Compare them with the reported approximately 60 kDa band. The supplied features do not establish alternative KRT6A isoforms, glycosylation sites, or a specific cause of extra bands, so verify their identity before assigning them to KRT6A.
Boster reagents
KRT6A 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 6 Rabbit Monoclonal Antibody
Cat # M30943-1
The catalog reports one human-reactive anti-Cytokeratin 6 antibody, M30943-1. Its Western blot image shows a reported band near 60 kDa in human A431 and HaCaT whole-cell lysates. The supplied evidence covers those tested samples and conditions.
Which to pick: M30943-1 is the only listed option for KRT6A. Its Western blot image uses human A431 and HaCaT lysates under reducing conditions, with the primary antibody at 1:500. Suitability for other samples is not shown.
Source: BosterBio KRT6A gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P02538.
- Human Protein Atlas. KRT6A tissue expression.
- PMC7225834 — target-verified WB comparison
- PMC13359832 — target-verified WB comparison
- PMC5558496 — target-verified WB comparison