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- Table of Contents
TMIGD1 / Transmembrane and immunoglobulin domain-containing protein 1 · Western blot design guide
Design a Western Blot for TMIGD1
Source-linked TMIGD1 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-TMIGD1 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
TMIGD1 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 | ~29.2 kDa | |
| Observed band | 68 kDa | |
| Gel | 12–15% (standard starting point) | |
| Negative control | Suggested KO / knockdown lysate |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Glycosylated + Cleaved | |
| Caveat | Glycosylation-state controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 2 isoform(s) |
Section 1
Source-Linked TMIGD1 Western Blot Protocol Options
The A16616 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | rat liver tissue lysate (catalog A16616) |
| Gel % | 12–15% (standard starting point) |
| Load | 20–30 µg total protein per lane; optimize for abundance (standard starting point) |
| Transfer | Short semi-dry transfer; verify retention (standard starting point) |
| Membrane | 0.45 µm PVDF (standard starting point) |
| Blocking | 5% milk or 5% BSA in TBST (standard starting point) |
| Primary antibody | A16616 · (A) 1 and (B) 2 μg/ml (catalog A16616) |
| 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 TMIGD1 Western Blot Band Size?
TMIGD1 is predicted at 29.2 kDa, while an empirical blot band is 68 kDa; the cause of that difference is not established.
| Band at 68 kDa | Empirical TMIGD1 band; confirm identity with antibody controls |
| Band near 29.2 kDa | Near the predicted full-length precursor mass; identity requires confirmation |
| Higher band near twice the monomer size | Possible homodimer if it persists during electrophoresis |
| Band below the precursor position | Could reflect removal of the 1–29 signal peptide |
| Several bands at different positions | Isoforms 1 and 2 are possible contributors, but distinct migration is unestablished |
| Band above the unmodified precursor position | N-linked glycosylation could contribute; the size effect is unquantified |
💡Expected TMIGD1 appearanceTMIGD1 has a predicted 29.2 kDa precursor mass and an empirical 68 kDa band; the difference is unexplained by the supplied evidence, so confirm band identity with antibody controls.
| Predicted precursor mass | 29.2 kDa provides a sequence-based reference, not a validated migration position |
| N-linked glycosylation at Asn58, Asn83, Asn118, Asn158 and Asn190 | Could increase apparent size; occupancy and migration effects are not supplied |
| Homodimer formation | Could yield a higher band near twice the monomer size if preserved |
| Splice isoforms 1 and 2 | May differ in size; their masses and migration difference are not supplied |
| Signal peptide at residues 1–29 | Cleavage could make mature protein smaller than the full-length precursor |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | TMIGD1 is a membrane protein and may be poorly extracted | Check membrane protein recovery and a positive lysate control |
| Band higher than expected | Glycosylation or persistent homodimers could affect migration; neither establishes the 68 kDa identity | Compare reducing conditions and use an independent antibody or TMIGD1 depletion control |
| Band lower than expected | Signal peptide cleavage could reduce the precursor size | Check epitope location and confirm identity with an independent antibody |
| Broad smear instead of sharp band | Heterogeneous N-linked glycosylation is possible | Compare untreated and deglycosylated samples with appropriate controls |
| Multiple bands | Isoforms 1 and 2 or different processing states are possible | Compare antibodies targeting shared and isoform-specific regions if available |
| Weak or no signal | Incomplete recovery of membrane-associated TMIGD1 is possible | Check extraction, sample loading and a positive control |
Experiment evidence
Western blot images showing TMIGD1 band
A16616 · TMIGD1
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for TMIGD1 Western blot
🧪For positive controls for TMIGD1 in Western blot, you can use no HPA-supported tissue or cell sample because none was supplied.
Positive control: No high/medium HPA tissue identified
Negative control: Suggested KO / knockdown lysate
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside the samples.
⚠️Feasibility: HPA tissue controls cannot be selected without expression data; a TMIGD1 knockdown or KO can provide a negative control.
HPA tissue expression evidence for TMIGD1
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 |
|---|---|---|---|---|
| No high/medium HPA tissues identified in the supplied evidence. | ||||
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| No lower-expression tissue rows available in the supplied evidence. | ||||
Section 3
Advanced TMIGD1 Western Blot Tips
Deeper troubleshooting and optimisation questions for TMIGD1, answered from its protein features.
How should TMIGD1 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 TMIGD1 isoforms produce different bands?
Isoforms · UniProt lists isoforms 1 and 2. Relative to isoform 1, isoform 2 has DKTVG replaced by ALHEG at residues 214–218 and lacks residues 219–262. Check whether the antibody epitope is retained before assigning a band to either isoform; their annotated sequence difference does not establish their observed migration.
Which TMIGD1 glycosylation sites could affect band interpretation?
PTM · UniProt annotates N-linked glycosylation at Asn58, Asn83, Asn118, Asn158, and Asn190. These are UniProt sequence coordinates; antibody or publication numbering may differ. A glycosidase comparison can help assess whether glycosylation contributes to band position or breadth.
Does this guide establish induction of TMIGD1?
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 TMIGD1 Western blot?
Transfer · TMIGD1 is annotated as a single-pass type I membrane protein. Ensure the sample is adequately solubilized, then check transfer efficiency and adjust transfer conditions empirically. The supplied features do not specify a particular membrane, buffer, or transfer setting.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A16616 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 TMIGD1 bands be quantified?
Quantitation · Use a consistent band definition across samples and report which band was measured. TMIGD1 has two isoforms, five annotated N-linked glycosylation sites, and a homodimer annotation, so a band at one position cannot be assigned to a particular form from molecular weight alone.
Why might TMIGD1 migrate above its predicted 29.2 kDa mass?
Interpretation · TMIGD1 has five annotated N-linked glycosylation sites and a signal peptide at residues 1–29. These features can affect apparent mass, but the annotations alone do not establish a visible shift or explain the reported 68 kDa band. Compare treated and untreated samples to assess glycosylation's contribution.
TMIGD1 is annotated as a homodimer. Two 29.2 kDa chains total about 58.4 kDa before considering processing or glycosylation, but this does not establish that the reported 68 kDa band is a dimer. Compare band behavior across sample preparation conditions before making that assignment.
UniProt lists two disulfide bonds. Compare reducing and nonreducing sample conditions if band patterns differ; the annotation supports this check but does not predict a specific band position.
TMIGD1 is annotated as a homodimer. Two 29.2 kDa chains total about 58.4 kDa before considering processing or glycosylation, but this does not establish that the reported 68 kDa band is a dimer. Compare band behavior across sample preparation conditions before making that assignment.
UniProt lists two disulfide bonds. Compare reducing and nonreducing sample conditions if band patterns differ; the annotation supports this check but does not predict a specific band position.
Boster reagents
TMIGD1 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-TMIGD1 Antibody
Cat # A16616
A16616 is an anti-TMIGD1 antibody listed as reactive with human, mouse, and rat. Its supplied Western blot image uses rat liver tissue lysate at 1 and 2 μg/ml; the supplied evidence does not show human or mouse WB samples.
Which to pick: A16616 is the only listed option. It has a Western blot image from rat liver lysate at 1 and 2 μg/ml. For human or mouse samples, listed reactivity is available, but no corresponding WB image is supplied.
Source: BosterBio TMIGD1 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q6UXZ0.
- Human Protein Atlas. TMIGD1 tissue expression.
- PMC10333293 — target-verified WB comparison