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- Table of Contents
GBP5 / Guanylate-binding protein 5 · Western blot design guide
Design a Western Blot for GBP5
Source-linked GBP5 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-GBP5 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
GBP5 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 | ~66.6 kDa | |
| Observed band | ≈67 kDa | |
| Gel | 10% (catalog A07110-1) | |
| Positive control | Gallbladder (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 | Methylated + Cleaved | |
| Caveat | Processing-state controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 2 isoform(s) |
Section 1
Source-Linked GBP5 Western Blot Protocol Options
The A07110-1 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human Jurkat, human U937, human U251 (catalog A07110-1) |
| Gel % | 10% (catalog A07110-1) |
| Load | 30 ug; reducing conditions (catalog A07110-1) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog A07110-1) |
| Membrane | nitrocellulose membrane (catalog A07110-1) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog A07110-1) |
| Primary antibody | A07110-1 · 1:1000 (catalog A07110-1) |
| Primary incubation | overnight at 4°C (catalog A07110-1) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog A07110-1) |
| Secondary incubation | 1.5 hour at RT (catalog A07110-1) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog A07110-1) |
| Detection | ECL (catalog A07110-1) |
Section 2
What Is the Expected GBP5 Western Blot Band Size?
GBP5 is predicted at 66.6 kDa and observed near 67 kDa; no cause for their small difference is established.
| Band at approximately 67 kDa | matches the empirical GBP5 band near its 66.6 kDa predicted mass |
| Band near 133 kDa | could reflect a GTP-dependent homodimer if it survives sample preparation |
| Band slightly below the precursor position | could reflect removal of the three-residue propeptide, though the shift may be unresolvable |
| Multiple nearby bands | could include isoforms 1 and 2, but their migration difference is unknown |
💡Expected GBP5 appearanceGBP5 has a predicted mass of 66.6 kDa and an empirical band at approximately 67 kDa; confirm band identity with appropriate controls if other bands appear.
| Predicted GBP5 mass | 66.6 kDa predicts a band near the observed 67 kDa |
| GTP-dependent homodimer | could appear near twice the monomer mass if the dimer survives sample preparation |
| Isoforms 1 and 2 | may differ in size, but their masses and migration difference are unspecified |
| Propeptide at residues 584–586 | cleavage would slightly reduce mass, with no established resolvable shift |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | GBP5 associates with cytoplasmic vesicle and Golgi membranes | check a membrane-enriched fraction alongside whole-cell lysate |
| Band higher than expected | a GTP-dependent homodimer may persist during sample preparation | compare denaturing preparations and verify GBP5 identity |
| Band lower than expected | propeptide cleavage removes only three residues and cannot explain a large shift | check antibody specificity and sample integrity |
| Multiple bands | isoforms 1 and 2 are annotated, but distinct migration is unproven | confirm band identity with isoform-aware controls |
| Weak or no signal | membrane-associated GBP5 may be underrepresented in the sampled fraction | assess membrane recovery and include a positive lysate control |
Experiment evidence
Western blot images showing GBP5 band
A07110-1 · GBP5
A07110 · GBP5
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for GBP5 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for GBP5 in Western blot, you can use gallbladder tissue, an HPA positive candidate.
Positive control: Gallbladder (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.
⚠️Feasibility: GBP5 is membrane-associated, so use lysis conditions that recover membrane-bound protein.
HPA tissue expression evidence for GBP5
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 |
|---|---|---|---|---|
| Gallbladder | glandular cells | Medium | Protein (IHC) | HPA → |
| Lung | macrophages | Medium | Protein (IHC) | HPA → |
| Spleen | cells in red pulp | Medium | Protein (IHC) | HPA → |
| Tonsil | non-germinal center cells | Medium | Protein (IHC) | HPA → |
| Urinary bladder | urothelial 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 → |
| Breast | adipocytes | Not detected | Protein (IHC) | HPA → |
| Bronchus | respiratory epithelial cells | Not detected | Protein (IHC) | HPA → |
| Caudate | glial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced GBP5 Western Blot Tips
Deeper troubleshooting and optimisation questions for GBP5, answered from its protein features.
How should GBP5 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 GBP5 isoforms produce bands of different sizes?
Isoforms · Yes. Isoform 2 has E489K and lacks residues 490–586 relative to isoform 1, so it has a shorter sequence. Check which isoforms the antibody epitope can detect before assigning a smaller band.
Check the antibody epitope: residues 490–586 are absent from isoform 2, while residues 1–488 are shared. The listed sequence differences support an isoform assignment only if the antibody detects the relevant region and the band sizes are consistent.
Check the antibody epitope: residues 490–586 are absent from isoform 2, while residues 1–488 are shared. The listed sequence differences support an isoform assignment only if the antibody detects the relevant region and the band sizes are consistent.
Which GBP5 modification should I consider when interpreting bands?
PTM · UniProt lists a cysteine methyl ester at position 583. Isoform 2 lacks this region. These are UniProt sequence coordinates; antibody or paper numbering may differ. The listed modification alone does not establish a visible band shift.
Does this guide establish induction of GBP5?
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 GBP5?
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 A07110-1 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 GBP5 be quantified?
Quantitation · Standard workflow guidance: quantify only a validated target band within the linear exposure range. Use consistent sample preparation and loading, retain biological replicates, and avoid interpreting saturation or loading differences as regulation.
Why is the GBP5 band near 67 kDa?
Interpretation · The observed band at approximately 67 kDa is close to the predicted 66.6 kDa. The listed features do not establish a visible shift or explain a difference between apparent and calculated mass.
UniProt lists a propeptide at residues 584–586. Keep this feature in mind when assessing a nearby band, but the feature alone does not establish that processing occurred in your sample or caused a visible shift.
Use an antibody with a known epitope and state whether it can detect both isoforms. GBP5 is listed at Golgi and cytoplasmic vesicle membranes with lipid anchoring, so keep sample preparation and the material compared consistent across lanes.
UniProt lists a propeptide at residues 584–586. Keep this feature in mind when assessing a nearby band, but the feature alone does not establish that processing occurred in your sample or caused a visible shift.
Use an antibody with a known epitope and state whether it can detect both isoforms. GBP5 is listed at Golgi and cytoplasmic vesicle membranes with lipid anchoring, so keep sample preparation and the material compared consistent across lanes.
Boster reagents
GBP5 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-GBP5 Antibody
Cat # A07110-1
Real WB data
Anti-Guanylate-binding protein 5 GBP5 Antibody
Cat # A07110
Two the supplier anti-GBP5 antibodies have supplied WB images. A07110-1 shows an approximately 67 kDa band in human cell and mouse cell or tissue lysates. A07110 shows rat liver lysate with and without blocking peptide. These are specific tested contexts.
Which to pick: For the documented human or mouse samples, consider A07110-1 (1:1000; Jurkat, U937, U251, spleen, and RAW264.7). For rat liver, consider A07110 (1 μg/mL; blocking-peptide comparison). Both have WB images; choose based on the sample you plan to test.
Source: BosterBio GBP5 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q96PP8.
- Human Protein Atlas. GBP5 tissue expression.
- PMC9561824 — target-verified WB comparison
- PMC11889220 — target-verified WB comparison
- PMC9201962 — target-verified WB comparison