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PTK7 / Inactive tyrosine-protein kinase 7 · Western blot design guide
Design a Western Blot for PTK7
Source-linked PTK7 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-PTK7 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
PTK7 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 | ~118.4 kDa | |
| Gel | 8–10% (standard starting point) | |
| Positive control | Endometrium (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 | Glycosylated + Phosphorylated | |
| Caveat | Modification-state controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 6 isoform(s) |
Section 1
Source-Linked PTK7 Western Blot Protocol Options
The A02957-1 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | (A) human colon, (B) mouse kidney and (C) rat liver tissue lysate (catalog A02957-1) |
| Gel % | 8–10% (standard starting point) |
| Load | 20–30 µg total protein per lane; optimize for abundance (standard starting point) |
| Transfer | Wet/tank transfer; optimize duration (standard starting point) |
| Membrane | 0.45 µm PVDF (standard starting point) |
| Blocking | 5% milk or 5% BSA in TBST (standard starting point) |
| Primary antibody | A02957-1 · 1 μg/mL (catalog A02957-1) |
| 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 PTK7 Western Blot Band Size?
PTK7's predicted precursor mass is 118.4 kDa; glycosylation, signal-peptide cleavage, and isoforms could affect migration, but no empirical band size or feature-specific shift is established.
| Band near 118.4 kDa | Compatible with the predicted PTK7 precursor mass; identity requires controls |
| Band above 118.4 kDa | Could reflect N-linked glycosylation, but its migration effect is unconfirmed |
| Band below 118.4 kDa | Could reflect signal-peptide cleavage or an isoform; band identity requires controls |
| Several bands at different positions | Could include PTK7 isoforms 1–6, though distinct migration is unconfirmed |
💡Expected PTK7 appearancePTK7 has a predicted precursor mass of 118.4 kDa; signal-peptide cleavage, N-linked glycosylation, and isoforms may affect migration, but no empirical band size is supplied, so confirm identity with controls.
| Predicted PTK7 precursor mass | Provides a sequence-based reference of 118.4 kDa, not a measured band position |
| N-linked glycosylation at Asn116, Asn175, Asn184, Asn214, Asn268, Asn283, Asn405, and Asn463 | May increase apparent size; no shift magnitude is established |
| Signal peptide at residues 1–30 | Cleavage may make mature PTK7 smaller than the precursor |
| Splice isoforms 1, 2, 3, 4, 5, and 6 | May differ in size; their masses and resolvable band positions are unspecified |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | PTK7 is a membrane protein and may be poorly recovered | Check membrane extraction and use a positive-control lysate |
| Band higher than expected | N-linked glycosylation may alter migration | Compare matched samples before and after deglycosylation |
| Band lower than expected | Signal-peptide cleavage or an isoform may change size | Check antibody epitope coverage and confirm identity with a second antibody |
| Broad smear instead of sharp band | Heterogeneous N-linked glycosylation is possible | Compare matched samples before and after deglycosylation |
| Multiple bands | Isoforms 1–6 are annotated, but distinct bands are unconfirmed | Compare isoform expression and verify bands with PTK7 knockdown |
| Weak or no signal | Recovery of membrane-localized PTK7 may be low | Check membrane extraction and include a positive-control lysate |
Experiment evidence
Western blot images showing PTK7 band
A02957-1 · PTK7
A02957 · PTK7
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for PTK7 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for PTK7 in Western blot, you can use endometrium tissue, which has high HPA expression.
Positive control: Endometrium (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: PTK7 is a membrane protein, so efficient membrane protein extraction may be needed for a clear signal.
HPA tissue expression evidence for PTK7
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 |
|---|---|---|---|---|
| Endometrium | cells in endometrial stroma | High | Protein (IHC) | HPA → |
| Breast | glandular cells | Medium | Protein (IHC) | HPA → |
| Bronchus | respiratory epithelial cells | Medium | Protein (IHC) | HPA → |
| Caudate | neuronal cells | Medium | Protein (IHC) | HPA → |
| Cerebellum | Purkinje 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 → |
| Appendix | glandular cells | Not detected | Protein (IHC) | HPA → |
| Bone marrow | hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Heart muscle | cardiomyocytes | Not detected | Protein (IHC) | HPA → |
| Parathyroid gland | glandular cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced PTK7 Western Blot Tips
Deeper troubleshooting and optimisation questions for PTK7, answered from its protein features.
How should PTK7 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 PTK7 isoforms produce bands at different sizes?
Isoforms · Six isoforms are listed. Relative to the canonical sequence, isoform 2 lacks residues 500–539, isoform 4 lacks 627–682, and isoform 5 lacks 817–1070. Isoform 3 replaces residues 410–540 with one residue. Check whether the antibody epitope is retained before assigning a band to an isoform.
What do PTK7 glycosylation sites mean for band interpretation?
PTM · Ten N-linked sites are annotated at UniProt positions 116, 175, 184, 214, 268, 283, 405, 463, 567, and 646. They make glycosylation relevant when assessing mobility, but the annotations alone cannot predict a band position or prove that glycosylation explains an unexpected band.
UniProt lists phosphoserine at position 1064 in the canonical sequence. Isoform 5 lacks residues 817–1070, so it lacks this site. If comparing with antibody or paper numbering, check which isoform and sequence convention they use. This annotation alone does not establish a detectable band shift.
UniProt lists phosphoserine at position 1064 in the canonical sequence. Isoform 5 lacks residues 817–1070, so it lacks this site. If comparing with antibody or paper numbering, check which isoform and sequence convention they use. This annotation alone does not establish a detectable band shift.
Does this guide establish induction of PTK7?
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 PTK7 Western blot?
Transfer · PTK7 is a predicted 118.4 kDa, single-pass membrane protein. Use transfer conditions suitable for a protein of that size and verify transfer with a membrane stain or marker. The supplied features do not establish that one transfer method is best.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A02957-1 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 multiple PTK7 bands be quantified?
Quantitation · Define which band or bands are being measured and use the same choice across samples. PTK7 has six isoforms, including isoform 5 with a large C-terminal deletion; antibody epitope location affects which forms can be detected. The supplied features do not identify any observed band, so avoid treating every signal as the same PTK7 form.
Why might PTK7 migrate differently from its predicted 118.4 kDa?
Interpretation · PTK7 has a signal peptide at residues 1–30 and ten annotated N-linked glycosylation sites. These features may affect apparent mass, but they do not establish a visible shift. No observed band size is supplied, so the predicted mass cannot be reconciled with a specific band.
First compare the antibody epitope with the six isoforms and their altered regions. Consider the ten N-linked glycosylation sites and signal peptide at residues 1–30 when assessing mobility. None of these features alone identifies an unexpected band; an empirical band size and further validation are needed.
First compare the antibody epitope with the six isoforms and their altered regions. Consider the ten N-linked glycosylation sites and signal peptide at residues 1–30 when assessing mobility. None of these features alone identifies an unexpected band; an empirical band size and further validation are needed.
Boster reagents
PTK7 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-PTK7 Antibody
Cat # A02957-1
Real WB data
Anti-CCK-4 PTK7 Monoclonal Antibody
Cat # A02957
Both listed anti-PTK7 antibodies have Western blot images. A02957-1 was shown with human colon, mouse kidney, and rat liver lysates; A02957 was shown with six human cell line lysates. No publication evidence is supplied.
Which to pick: For mouse or rat samples, consider A02957-1, which lists those species and shows tissue lysate blots. For human cell lysates, consider A02957, which lists human reactivity and shows blots of HeLa, A431, HCT116, Caco2, HepG2, and MCF-7 lysates.
Source: BosterBio PTK7 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q13308.
- Human Protein Atlas. PTK7 tissue expression.
- PMC12130886 — target-verified WB comparison
- PMC5445388 — target-verified WB comparison
- PMC3883666 — target-verified WB comparison