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- Table of Contents
GZF1 / GDNF-inducible zinc finger protein 1 · Western blot design guide
Design a Western Blot for GZF1
Source-linked GZF1 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-GZF1 WB antibodies. Everything you need to plan the experiment before you commit precious samples.
Evidence assembled October 2026 · For research use; verify linked source records and product datasheet before use
GZF1 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 | ~80.5 kDa | |
| Observed band | 68 kDa | |
| Gel | 8–10% (standard starting point) | |
| Positive control | Adrenal gland (IHC candidate; verify WB) +4 more | |
| Negative control | Appendix (IHC candidate; verify WB) |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Phosphorylated | |
| Caveat | Phosphorylation-state controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 2 isoform(s) |
Section 1
Source-Linked GZF1 Western Blot Protocol Options
The A11916 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human heart tissue lysate (catalog A11916) |
| Gel % | 8–10% (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 | 5% milk or 5% BSA in TBST (standard starting point) |
| Primary antibody | A11916 · 1 μg/mL (catalog A11916) |
| 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 GZF1 Western Blot Band Size?
GZF1 is predicted at 80.5 kDa, while antibody QC reports 68 kDa; the cause of the difference is not established.
| Band near 68 kDa | Empirical GZF1 antibody band; confirm identity with controls |
| Band near 80.5 kDa | Near the UniProt predicted mass, with identity requiring confirmation |
| Several bands at different positions | Isoforms 1 and 2 are annotated, but distinct migration is unverified |
| Weak signal in a cytoplasmic fraction | GZF1 is also found in the nucleus |
💡Expected GZF1 appearanceUniProt predicts 80.5 kDa, while antibody QC reports a 68 kDa band in human heart lysate; the reason for the difference is unestablished, so confirm band identity with appropriate controls.
| UniProt predicted mass | 80.5 kDa is the sequence-based reference |
| Isoform 1 | Its individual mass and migration are not supplied |
| Isoform 2 | Alternative splicing may change size, but its individual mass and migration are not supplied |
| Phosphoserine 613 | A modification is annotated, but an apparent size effect is not established |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | GZF1 may be poorly recovered if nuclear proteins are underextracted | Check nuclear protein recovery and a positive lysate |
| Band higher than expected | Band identity or migration is uncertain | Compare with a positive control and confirm specificity by GZF1 depletion |
| Band lower than expected | Antibody QC reports 68 kDa despite the 80.5 kDa prediction; the cause is unknown | Compare with the QC sample and confirm specificity by GZF1 depletion |
| Multiple bands | Two isoforms are annotated, but distinct bands are unverified | Use GZF1 depletion to identify specific bands |
| Weak or no signal | Recovery may depend on nuclear extraction | Check extraction and compare cytoplasmic and nuclear fractions |
Experiment evidence
Western blot images showing GZF1 band
A11916 · GZF1
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for GZF1 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for GZF1 in Western blot, you can use adrenal gland lysate, based on its high HPA expression.
Positive control: Adrenal gland (IHC candidate; verify WB)
Negative control: Appendix (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain such as Ponceau alongside the samples.
⚠️Feasibility: Appendix provides a plausible negative tissue, though HPA non-detection does not establish a true null.
HPA tissue expression evidence for GZF1
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 |
|---|---|---|---|---|
| Adrenal gland | glandular cells | High | Protein (IHC) | HPA → |
| Skeletal muscle | myocytes | High | Protein (IHC) | HPA → |
| Colon | endothelial cells | Medium | Protein (IHC) | HPA → |
| Endometrium | glandular cells | Medium | Protein (IHC) | HPA → |
| Gallbladder | glandular cells | Medium | Protein (IHC) | HPA → |
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Appendix | glandular cells | Not detected | Protein (IHC) | HPA → |
| Bone marrow | hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Bronchus | respiratory epithelial 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 GZF1 Western Blot Tips
Deeper troubleshooting and optimisation questions for GZF1, answered from its protein features.
How should GZF1 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.
How could GZF1 isoforms affect the blot?
Isoforms · UniProt lists isoforms 1 and 2; isoform 2 lacks residues 1–491 of the canonical sequence. An antibody targeting that missing region would not detect isoform 2. The supplied features do not assign the 68 kDa band to either isoform.
Does phosphorylation explain a shifted GZF1 band?
PTM · UniProt lists phosphoserine at position 613 in the canonical sequence. This site alone does not establish a visible shift. If testing phosphorylation, compare samples with and without phosphatase treatment; keep UniProt’s canonical numbering explicit when comparing antibody or paper coordinates.
Does this guide establish induction of GZF1?
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 GZF1 Western blot?
Transfer · The supplied features do not specify a transfer method. Assess recovery around the predicted 80.5 kDa mass and the reported 68 kDa band with your chosen setup; neither value alone identifies an optimal transfer method.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A11916 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 GZF1 be quantified across cell fractions?
Quantitation · GZF1 is reported in cytoplasm, nucleoplasm and nucleolus, and its nuclear localization depends on NCL. Quantify comparable fractions separately and account for their loading when comparing samples; a changed nuclear signal could reflect localization rather than a change in total GZF1.
Why might GZF1 run at 68 kDa instead of 80.5 kDa?
Interpretation · The reported apparent band is 68 kDa, while the predicted mass is 80.5 kDa. The supplied features do not establish why they differ. Check the band’s identity before assigning the difference to phosphorylation or an isoform.
Consider antibody recognition of the two isoforms: isoform 2 lacks canonical residues 1–491. UniProt also lists phosphoserine 613, but does not show that it produces a distinct band. Compare antibody epitope coverage and fraction-specific patterns before assigning an unexpected band.
Consider antibody recognition of the two isoforms: isoform 2 lacks canonical residues 1–491. UniProt also lists phosphoserine 613, but does not show that it produces a distinct band. Compare antibody epitope coverage and fraction-specific patterns before assigning an unexpected band.
Boster reagents
GZF1 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-GZF1 Antibody
Cat # A11916
the supplier A11916 is an anti-GZF1 antibody listed for human reactivity. Its Western blot image uses human heart tissue lysate at 1 μg/mL. No publication evidence or other tested specimens are supplied.
Which to pick: A11916 is the only listed option. Its documented Western blot example uses human heart tissue lysate; evidence for other specimens is not provided.
Source: BosterBio GZF1 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q9H116.
- Human Protein Atlas. GZF1 tissue expression.
- PMC6156078 — target-verified WB comparison