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- Table of Contents
TGM1 / Protein-glutamine gamma-glutamyltransferase K · Western blot design guide
Design a Western Blot for TGM1
Source-linked TGM1 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-TGM1 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
TGM1 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 | ~89.8 kDa | |
| Gel | 8–10% (standard starting point) | |
| Positive control | Cervix (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 | Phosphorylated | |
| Caveat | Phosphorylation-state controls | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 2 isoform(s) |
Section 1
Source-Linked TGM1 Western Blot Protocol Options
The A02484 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | extracts of various cell lines, (catalog A02484) |
| 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 | A02484; 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 TGM1 Western Blot Band Size?
TGM1 has a predicted mass of 89.8 kDa; isoforms could affect migration, but no empirical band or distinct isoform pattern is established.
| Band near 89.8 kDa | consistent with the predicted full-length mass; confirm identity with controls |
| Two bands at different positions | could reflect isoforms 1 and 2, though distinct migration is unproven |
| Band in a membrane fraction | consistent with the reported membrane localization |
| Band in a cytosolic fraction | consistent with the reported cytosolic localization |
💡Expected TGM1 appearanceUniProt predicts 89.8 kDa for TGM1; no empirical band size is supplied, and isoform-dependent migration is unestablished, so confirm any candidate band with ordinary identity controls.
| UniProt predicted mass | full-length sequence has a calculated mass of 89.8 kDa |
| Splice isoform 1 | its individual apparent size is not supplied |
| Splice isoform 2 | its size relative to isoform 1 is not supplied |
| Alternative splicing of isoforms 1 and 2 | may yield different sizes, but distinct bands are unproven |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | membrane-associated TGM1 may be poorly recovered during extraction | check membrane extraction and use a positive control |
| Band higher than expected | isoform identity or anomalous migration is unresolved | compare a positive control and verify band identity |
| Band lower than expected | isoform identity or band specificity is unresolved | compare a positive control and verify band identity |
| Multiple bands | isoforms 1 and 2 are reported, but their migration is unknown | check each band with a specificity control |
| Weak or no signal | TGM1 recovery or detection may be insufficient | check lysate recovery and antibody performance with a positive control |
Experiment evidence
Western blot images showing TGM1 band
A02484 · TGM1
A02484 · TGM1
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for TGM1 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for TGM1 in Western blot, you can use cervix tissue, which HPA lists as a positive candidate.
Positive control: Cervix (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: HPA provides a tissue contrast, though TGM1’s membrane association may affect lysate recovery.
HPA tissue expression evidence for TGM1
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 |
|---|---|---|---|---|
| Cervix | squamous epithelial cells | Medium | Protein (IHC) | HPA → |
| Esophagus | squamous epithelial cells | Medium | Protein (IHC) | HPA → |
| Skin | cells in corneal layer | Medium | Protein (IHC) | HPA → |
| Tonsil | squamous epithelial 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 TGM1 Western Blot Tips
Deeper troubleshooting and optimisation questions for TGM1, answered from its protein features.
How should TGM1 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 TGM1 isoforms affect the blot?
Isoforms · UniProt lists isoforms 1 and 2; isoform 2 lacks residues 1–442. An antibody targeting that region would miss isoform 2. Check the antibody epitope before assigning a smaller band to an isoform.
Which TGM1 phosphorylation sites matter when interpreting bands?
PTM · The supplied UniProt coordinates are Thr22 and Ser24, Ser68, Ser82, Ser85, Ser92, and Ser95. Keep this coordinate convention when comparing antibody or paper numbering. Their presence does not establish a visible band shift; use a phosphosite-specific antibody if the question is whether a particular site is phosphorylated.
Does this guide establish induction of TGM1?
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 TGM1 Western blot?
Transfer · The supplied features do not specify a transfer method. For the predicted 89.8-kDa canonical protein, check transfer efficiency near that size with a total-protein stain. Membrane and cytosolic localization also make the fraction loaded relevant to the result.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A02484 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 can TGM1 be quantified across samples?
Quantitation · Compare the same sample fraction across conditions, since TGM1 occurs at the membrane and in the cytosol. Keep detection in a linear range and normalize to total protein in that fraction. An antibody against residues 1–442 will not measure isoform 2, which lacks that region.
Why might a TGM1 band differ from its predicted mass?
Interpretation · The canonical sequence predicts 89.8 kDa. TGM1 has two isoforms and seven listed phosphorylation sites, but these features alone do not establish a visible shift or explain any particular apparent mass. No observed band size was supplied; compare bands with the predicted mass and check which isoform the antibody recognizes.
TGM1 is reported at the cell membrane through lipid anchoring and in the cytosol. Myristoylation and palmitoylation mediate membrane localization. If analyzing a fraction, check whether it contains the compartment you intend to measure before interpreting a weak band.
Check band size against the 89.8-kDa canonical prediction, then check the antibody epitope: isoform 2 lacks residues 1–442. The listed phosphorylation sites alone do not identify an extra band. Compare membrane and cytosolic fractions when localization may affect which TGM1 species are present.
TGM1 is reported at the cell membrane through lipid anchoring and in the cytosol. Myristoylation and palmitoylation mediate membrane localization. If analyzing a fraction, check whether it contains the compartment you intend to measure before interpreting a weak band.
Check band size against the 89.8-kDa canonical prediction, then check the antibody epitope: isoform 2 lacks residues 1–442. The listed phosphorylation sites alone do not identify an extra band. Compare membrane and cytosolic fractions when localization may affect which TGM1 species are present.
Boster reagents
TGM1 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-TGM1 Antibody
Cat # A02484
The catalog reports Anti-TGM1 Antibody A02484 for Western blot, with stated human and rat reactivity. A WB image shows extracts from various cell lines, but the supplied caption does not identify the lines or provide testing conditions.
Which to pick: A02484 is the only listed option. Check its stated human and rat reactivity against your sample; the supplied WB image documents cell-line extracts without identifying the lines.
Source: BosterBio TGM1 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P22735.
- Human Protein Atlas. TGM1 tissue expression.
- PMC12676209 — target-verified WB comparison
- PMC3777415 — target-verified WB comparison
- PMC10700374 — target-verified WB comparison