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- Table of Contents
KLF7 / Krueppel-like factor 7 · Western blot design guide
Design a Western Blot for KLF7
Source-linked KLF7 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-KLF7 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
KLF7 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 | ~33.4 kDa | |
| Gel | 12–15% (standard starting point) | |
| Positive control | Appendix (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 | Peptide-blocking control | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 6 isoform(s) |
Section 1
Source-Linked KLF7 Western Blot Protocol Options
The A06749 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | Target-positive lysate and matched negative control (standard starting point) |
| Gel % | 12–15% (standard starting point) |
| Load | 20–30 µg total protein per lane; optimize for abundance (standard starting point) |
| Transfer | Standard semi-dry transfer; verify efficiency (standard starting point) |
| Membrane | 0.45 µm PVDF (standard starting point) |
| Blocking | the KLF7 peptide (catalog A06749) |
| Primary antibody | A06749; use the WB datasheet starting dilution (standard starting point) |
| 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 KLF7 Western Blot Band Size?
KLF7 is predicted at 33.4 kDa; six isoforms could affect migration, but no apparent band size or isoform separation is demonstrated.
| Band near 33.4 kDa | Consistent with the predicted KLF7 mass; identity requires confirmation. |
| Several bands at different positions | Could reflect KLF7 isoforms 1–6; distinct migration is unconfirmed. |
| Weak band in whole-cell lysate | Nuclear localization may limit the signal in an unfractionated sample. |
| Stronger band in a nuclear fraction | Consistent with KLF7's nuclear location; confirm band identity. |
💡Expected KLF7 appearanceKLF7 has a predicted mass of 33.4 kDa, but no empirical band size or isoform-specific migration is supplied; confirm a candidate band with nuclear fractionation and peptide blocking.
| Predicted KLF7 mass | Places the reference band near 33.4 kDa. |
| Splice isoforms 1 and 2 | May differ in apparent size; individual masses are unknown. |
| Splice isoforms 3 and 4 | May differ in apparent size; individual masses are unknown. |
| Splice isoforms 5 and 6 | May differ in apparent size; individual masses are unknown. |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Nuclear KLF7 may be underrepresented in whole-cell lysate. | Check a nuclear fraction and verify recovery with a nuclear marker. |
| Band higher than expected | An isoform is possible, but its migration is unknown. | Compare with peptide-blocked antibody signal and a molecular-weight marker. |
| Band lower than expected | An isoform is possible, but its migration is unknown. | Check peptide blocking and compare a nuclear fraction. |
| Multiple bands | Six KLF7 isoforms are listed, but distinct bands are unconfirmed. | Check which bands disappear with peptide blocking. |
| Weak or no signal | Nuclear localization may reduce signal in the sampled fraction. | Assess nuclear extraction and sample loading. |
Experiment evidence
Western blot images showing KLF7 band
A06749 · KLF7
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for KLF7 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for KLF7 in Western blot, you can use appendix tissue, which shows high HPA expression.
Positive control: Appendix (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: Tissue controls are feasible, but nuclear enrichment may improve KLF7 detection.
HPA tissue expression evidence for KLF7
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 |
|---|---|---|---|---|
| Appendix | lymphoid tissue | High | Protein (IHC) | HPA → |
| Colon | glandular cells | High | Protein (IHC) | HPA → |
| Duodenum | glandular cells | High | Protein (IHC) | HPA → |
| Rectum | glandular cells | High | Protein (IHC) | HPA → |
| Small intestine | glandular 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 → |
| Liver | cholangiocytes | Not detected | Protein (IHC) | HPA → |
| Ovary | ovarian stroma cells | Not detected | Protein (IHC) | HPA → |
| Skeletal muscle | myocytes | Not detected | Protein (IHC) | HPA → |
| Smooth muscle | smooth muscle cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced KLF7 Western Blot Tips
Deeper troubleshooting and optimisation questions for KLF7, answered from its protein features.
Where should the canonical KLF7 band appear?
Band shift · Canonical KLF7 is 302 amino acids with a predicted mass of 33.4 kDa. No observed band position is supplied, so use 33.4 kDa as a starting reference, not a confirmed migration position.
Can KLF7 isoforms produce bands of different sizes?
Isoforms · Six isoforms are listed. UniProt reports deletions of residues 55–244 in isoform 5 and 88–302 in isoform 6, plus a replacement of 179–302 in isoform 3. These sequence changes can alter predicted size, but the features alone do not establish visible bands or their apparent masses. Coordinates refer to the supplied canonical UniProt sequence.
Detection depends on the antibody epitope. Isoform 5 lacks canonical residues 55–244; isoform 6 lacks 88–302; and isoform 3 replaces 179–302. Check the epitope against each isoform sequence before treating a missing band as absent KLF7. Coordinates use the supplied canonical UniProt sequence.
Possibly, if their epitopes cover the changed sequence. Isoform 2 replaces canonical residues 1–34 with M, while isoform 4 replaces them with MFPSWP. An antibody against the canonical N terminus may therefore recognize these isoforms differently. Coordinates use the supplied canonical UniProt sequence.
Detection depends on the antibody epitope. Isoform 5 lacks canonical residues 55–244; isoform 6 lacks 88–302; and isoform 3 replaces 179–302. Check the epitope against each isoform sequence before treating a missing band as absent KLF7. Coordinates use the supplied canonical UniProt sequence.
Possibly, if their epitopes cover the changed sequence. Isoform 2 replaces canonical residues 1–34 with M, while isoform 4 replaces them with MFPSWP. An antibody against the canonical N terminus may therefore recognize these isoforms differently. Coordinates use the supplied canonical UniProt sequence.
Do annotated modifications prove a band shift?
PTM · The linked UniProt record describes protein features. A modification annotation alone does not demonstrate a visible shift; retain any condition or experimental qualifier attached to it.
Does this guide establish induction of KLF7?
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 KLF7?
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 A06749 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 KLF7 bands be quantified across samples?
Quantitation · Quantify the same clearly defined band across comparable samples and fractions. Because six isoforms are listed and some have substantial sequence changes, avoid summing bands or comparing different bands as though they represent the same KLF7 product without identifying them first.
Which sample fraction is relevant for KLF7 detection?
Interpretation · UniProt places KLF7 in the nucleus. Include a nuclear fraction when assessing whether the protein is detectable, and keep the fraction consistent across samples being compared.
First compare each band with the six listed isoform sequences and the antibody epitope. Sequence changes make different products plausible, but no observed band positions or modified residues are supplied. A band's position alone cannot identify an isoform or establish a modification.
First compare each band with the six listed isoform sequences and the antibody epitope. Sequence changes make different products plausible, but no observed band positions or modified residues are supplied. A band's position alone cannot identify an isoform or establish a modification.
Boster reagents
KLF7 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-Krueppel-like factor 7 KLF7 Antibody
Cat # A06749
A06749 is an anti-KLF7 antibody listed as reactive with Human and Mouse. Its Western blot image includes a right lane blocked with KLF7 peptide. The supplied caption does not identify the sample or establish broader validation.
Which to pick: A06749 is the only listed antibody. Its stated Human and Mouse reactivity and peptide-blocked Western blot image are the available evidence to consider when choosing it for your sample.
Source: BosterBio KLF7 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry O75840.
- Human Protein Atlas. KLF7 tissue expression.
- PMC11299624 — target-verified WB comparison
- PMC12924756 — target-verified WB comparison
- PMC8107552 — target-verified WB comparison