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- Table of Contents
GGT1 / Glutathione hydrolase 1 proenzyme · Western blot design guide
Design a Western Blot for GGT1
Source-linked GGT1 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-GGT1 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
GGT1 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 | ~61.4 kDa | |
| Gel | 12–15% (standard starting point) | |
| Positive control | Kidney (IHC candidate; verify WB) +3 more | |
| Negative control | Adipose tissue (IHC candidate; verify WB) |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Glycosylated | |
| Caveat | Modification-state controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 3 isoform(s) |
Section 1
Source-Linked GGT1 Western Blot Protocol Options
The M02340 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | Human fetal liver lysate (catalog M02340) |
| 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 | 5% milk or 5% BSA in TBST (standard starting point) |
| Primary antibody | M02340; 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 GGT1 Western Blot Band Size?
GGT1 has a predicted 61.4 kDa proenzyme; glycosylation, heavy/light chains, and isoforms could affect bands, but their migration effects are unproven and no observed size is supplied.
| Band near 61.4 kDa | Compatible with the predicted proenzyme size; confirm identity with antibody controls. |
| Band below 61.4 kDa | Could represent an antibody-recognized heavy or light chain of the annotated heterodimer; chain masses are unavailable. |
| Several bands | Isoforms 1, 2, and 3 or heavy and light chains are possible contributors; distinct isoform migration is unproven. |
| Band above 61.4 kDa or diffuse signal | N-linked glycosylation may affect migration, but the annotated sites do not establish a visible shift or smear. |
| Weak signal in soluble lysate | GGT1 is a single-pass membrane protein and may be underrepresented in that fraction. |
💡Expected GGT1 appearanceUniProt predicts a 61.4 kDa proenzyme; annotated N-linked sites, heavy and light chains, and isoforms may alter the antibody-recognized pattern, but no empirical band size is supplied, so confirm identity with controls.
| Predicted proenzyme mass | Provides a 61.4 kDa sequence-based reference, not a validated band position. |
| N-linked sites at Asn95 and Asn120 | May affect apparent migration if glycosylated; no site-specific shift is established. |
| N-linked sites at Asn230, Asn266, and Asn297 | May affect apparent migration if glycosylated; no site-specific shift is established. |
| N-linked sites at Asn344 and Asn511 | May affect apparent migration if glycosylated; no site-specific shift is established. |
| Isoforms 1, 2, and 3 | May differ in size, but their masses and resolvable migration differences are not supplied. |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Membrane-bound GGT1 may be poorly recovered in the lysate. | Check a membrane-enriched fraction and confirm extraction and antibody performance with controls. |
| Band higher than expected | N-linked glycosylation could alter migration; the cause is unconfirmed. | Compare appropriate glycosylation controls and verify band identity with an independent antibody. |
| Band lower than expected | The antibody may recognize a heavy or light chain rather than the proenzyme. | Check the antibody epitope and compare bands with an independent GGT1 antibody. |
| Broad smear instead of sharp band | Variable N-linked glycosylation is possible but unverified. | Assess glycosylation with an appropriate enzymatic control and check sample integrity. |
| Multiple bands | Heavy and light chains or isoforms may contribute; distinct isoform bands are unproven. | Check epitope coverage and test band identity with an independent antibody. |
| Weak or no signal | Insufficient recovery of the single-pass membrane protein is possible. | Optimize membrane protein extraction and include a positive control. |
Experiment evidence
Western blot images showing GGT1 band
M02340 · GGT1
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for GGT1 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for GGT1 in Western blot, you can use kidney tissue, which has high HPA expression.
Positive control: Kidney (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: As GGT1 is a membrane protein, a membrane-enriched sample may improve detection.
HPA tissue expression evidence for GGT1
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 |
|---|---|---|---|---|
| Kidney | cells in tubules | High | Protein (IHC) | HPA → |
| Stomach | glandular cells | High | Protein (IHC) | HPA → |
| Epididymis | glandular 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 → |
| 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 GGT1 Western Blot Tips
Deeper troubleshooting and optimisation questions for GGT1, answered from its protein features.
How should GGT1 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 isoform 2 affect GGT1 band identification?
Isoforms · In canonical UniProt coordinates, isoform 2 replaces residues 341–366 and lacks residues 367–569. An antibody against the missing region would not recognize that isoform. Check the antibody epitope before assigning a band to isoform 2.
Isoform 3 lacks canonical UniProt residues 1–344. An antibody against that region would not recognize it. Check epitope coverage before interpreting a missing or additional band as isoform-specific.
Isoform 3 lacks canonical UniProt residues 1–344. An antibody against that region would not recognize it. Check epitope coverage before interpreting a missing or additional band as isoform-specific.
Which GGT1 glycosylation sites matter when interpreting bands?
PTM · The annotated N-linked sites are Asn 95, 120, 230, 266, 297, 344, and 511 in canonical UniProt numbering. If comparing antibody or paper site numbers, check their sequence convention first. Site annotation alone does not establish a visible band shift.
Does this guide establish induction of GGT1?
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 GGT1?
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 M02340 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 GGT1 bands be quantified across samples?
Quantitation · Define the band and antibody epitope consistently across samples. GGT1 has three isoforms, and the protein forms heavy and light chains; an antibody may detect only some forms. Avoid combining distinct bands into one measurement without establishing what each represents.
Why might GGT1 migrate differently from its predicted 61.4 kDa?
Interpretation · 61.4 kDa is the predicted mass of the full-length canonical sequence. GGT1 has seven annotated N-linked glycosylation sites and forms a heavy- and light-chain heterodimer. These features make a different apparent band plausible, but the supplied evidence gives no observed band size or demonstrated shift.
GGT1 is annotated as a heterodimer of heavy and light chains, with the active site in the light chain. Consider chain recognition when interpreting multiple bands, but the supplied features do not specify chain boundaries or expected band masses. Confirm which chain the antibody detects.
Check antibody epitope coverage against isoforms 2 and 3, and consider heavy- and light-chain recognition. GGT1 is also a glycoprotein with seven annotated N-linked sites. These features suggest possibilities to test, but they do not identify any particular unexpected band or establish its apparent mass.
GGT1 is annotated as a heterodimer of heavy and light chains, with the active site in the light chain. Consider chain recognition when interpreting multiple bands, but the supplied features do not specify chain boundaries or expected band masses. Confirm which chain the antibody detects.
Check antibody epitope coverage against isoforms 2 and 3, and consider heavy- and light-chain recognition. GGT1 is also a glycoprotein with seven annotated N-linked sites. These features suggest possibilities to test, but they do not identify any particular unexpected band or establish its apparent mass.
Boster reagents
GGT1 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-GGT1 Rabbit Monoclonal Antibody
Cat # M02340
The listed anti-GGT1 rabbit monoclonal antibody, M02340, is reported to react with human GGT1. Its Western blot image shows GGT1 expression in human fetal liver lysate; the supplied evidence does not establish performance in other samples.
Which to pick: M02340 is the only listed option. Choose it for a human GGT1 Western blot when the reported human fetal liver lysate example is relevant; confirm suitability for other sample types.
Source: BosterBio GGT1 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P19440.
- Human Protein Atlas. GGT1 tissue expression.
- PMC12117263 — target-verified WB comparison
- PMC10097650 — target-verified WB comparison
- PMC5420129 — target-verified WB comparison