This website uses cookies to ensure you get the best experience on our website.
- Table of Contents
TXNRD1 / Thioredoxin reductase 1, cytoplasmic · Western blot design guide
Design a Western Blot for TXNRD1
Real validated TXNRD1 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-TXNRD1 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
TXNRD1 Western Blot Experimental Design Guide
Expected bands, validated protocols, controls and antibodies — the at-a-glance facts below, then the full design guide.
Must know before running
| Expected band | ~70.9 kDa | |
| Observed band | ~65 kDa | |
| Gel | 5–20% (catalog A01778-3) | |
| Positive control | Adrenal gland (IHC candidate; verify WB) +4 more | |
| Negative control | Suggested KO / knockdown lysate |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Phosphorylated + Acetylated | |
| Caveat | Modification-state controls | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 7 isoform(s) |
Section 1
Real Curated TXNRD1 Western Blot Protocols
The A01778-3 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human Jurkat, human A549, human MCF-7 (catalog A01778-3) |
| Gel % | 5–20% (catalog A01778-3) |
| Load | 30 ug; reducing conditions (catalog A01778-3) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog A01778-3) |
| Membrane | nitrocellulose membrane (catalog A01778-3) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog A01778-3) |
| Primary antibody | A01778-3 · 0.5 μg/mL (catalog A01778-3) |
| Primary incubation | overnight at 4°C (catalog A01778-3) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog A01778-3) |
| Secondary incubation | 1.5 hour at RT (catalog A01778-3) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog A01778-3) |
| Detection | ECL (catalog A01778-3) |
Section 2
What Is the Expected TXNRD1 Western Blot Band Size?
TXNRD1 is predicted at 70.9 kDa, but antibody QC shows ~65 kDa in reducing lysates; the cause of this difference is not established.
| Band at ~65 kDa | Empirical TXNRD1 band in reducing whole-cell lysates; confirm identity with controls |
| Band near 70.9 kDa | Near the calculated monomer mass; identity requires confirmation |
| Band near twice the monomer size | Could reflect a homodimer if it survives sample preparation |
| Several bands at different sizes | Could reflect splice isoforms 1–7; their migration is not established |
💡Expected TXNRD1 appearanceTXNRD1 has a predicted mass of 70.9 kDa, while antibody QC shows a ~65 kDa band in reducing whole-cell lysates; the cause of the difference is unknown, so confirm identity with appropriate controls.
| Predicted monomer mass | Calculated at 70.9 kDa; the empirical whole-cell lysate band is ~65 kDa |
| Isoform 1 | May differ in size from other splice isoforms; its individual mass is not supplied |
| Isoforms 2–7 | May differ in size from isoform 1 or each other; individual masses are not supplied |
| Homodimer formation | Could produce a band near twice the monomer size if the dimer persists during electrophoresis |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Cytoplasmic TXNRD1 was not detected in the prepared lysate | Check lysate loading and detection with a positive whole-cell lysate |
| Band higher than expected | A TXNRD1 homodimer may persist during sample preparation | Compare reducing and nonreducing preparations and confirm band identity |
| Band lower than expected | The observed ~65 kDa band runs below the 70.9 kDa predicted mass for an unestablished reason | Compare with the reported whole-cell lysate band and verify identity by TXNRD1 depletion |
| Multiple bands | Splice isoforms 1–7 are documented, but distinct band sizes are unproven | Use TXNRD1 depletion to identify which bands depend on TXNRD1 |
| Fragments below expected size | Fragment identity is not established by the supplied features | Check sample integrity and confirm TXNRD1 dependence by depletion |
Experiment evidence
Western blot images showing TXNRD1 band
A01778-3 · TXNRD1
M01778 · TXNRD1
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for TXNRD1 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for TXNRD1 in Western blot, you can use adrenal gland tissue, which shows high HPA expression.
Positive control: Adrenal gland (IHC candidate; verify WB)
Negative control: Suggested KO / knockdown lysate
Loading controls: Run GAPDH, β-actin, and a total-protein stain such as Ponceau alongside samples.
⚠️Feasibility: No tissue is listed as not detected, so use siRNA knockdown or a KO line for a negative control.
HPA tissue expression evidence for TXNRD1
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 → |
| Caudate | neuronal cells | High | Protein (IHC) | HPA → |
| Cerebellum | cells in molecular layer | High | Protein (IHC) | HPA → |
| Cerebral cortex | neuronal cells | High | Protein (IHC) | HPA → |
| Endometrium | glandular cells | High | Protein (IHC) | HPA → |
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | adipocytes | Low | Protein (IHC) | HPA → |
| Breast | adipocytes | Low | Protein (IHC) | HPA → |
| Esophagus | squamous epithelial cells | Low | Protein (IHC) | HPA → |
| Oral mucosa | squamous epithelial cells | Low | Protein (IHC) | HPA → |
| Vagina | squamous epithelial cells | Low | Protein (IHC) | HPA → |
Section 3
Advanced TXNRD1 Western Blot Tips
Deeper troubleshooting and optimisation questions for TXNRD1, answered from its protein features.
How should TXNRD1 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 TXNRD1 isoforms produce additional bands?
Isoforms · Seven isoforms are listed. Isoforms 2, 3, 4, 5, and 7 have N-terminal deletions, and several have replacement sequences. These differences could affect migration, but the features do not establish which isoforms are present or detectable in your sample.
Check the epitope against each isoform you intend to measure. Isoforms 2, 3, 4, 5, and 7 lose portions of the canonical N terminus; isoforms 2, 3, 4, and 6 also have N-terminal sequence replacements. An antibody targeting an affected region may miss an isoform.
Decide whether the measurement represents one band or all confirmed TXNRD1 bands. Seven isoforms are listed, with several N-terminal differences, so an antibody may recognize them differently. Report the band or bands quantified and the antibody epitope when known.
Check the epitope against each isoform you intend to measure. Isoforms 2, 3, 4, 5, and 7 lose portions of the canonical N terminus; isoforms 2, 3, 4, and 6 also have N-terminal sequence replacements. An antibody targeting an affected region may miss an isoform.
Decide whether the measurement represents one band or all confirmed TXNRD1 bands. Seven isoforms are listed, with several N-terminal differences, so an antibody may recognize them differently. Report the band or bands quantified and the antibody epitope when known.
Which annotated modifications matter when interpreting TXNRD1 bands?
PTM · The supplied UniProt features list N6-succinyllysine at position 218 and an annotation reading 'Phosphotyrosine N-acetylmethionine' at position 281. These are UniProt coordinates; antibody or paper numbering may differ. Neither annotation alone demonstrates a visible shift or explains the approximately 65 kDa band.
Does this guide establish induction of TXNRD1?
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 TXNRD1?
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 A01778-3 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 TXNRD1 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 might TXNRD1 appear near 65 kDa instead of 70.9 kDa?
Interpretation · The supplied observed band is approximately 65 kDa, while the predicted mass is 70.9 kDa. These values differ, but the listed features do not establish the cause. Confirm the band with an appropriate TXNRD1 control before assigning it.
TXNRD1 is annotated as a homodimer, making dimerization relevant when evaluating a higher band. The annotation does not show that a dimer survives your Western blot conditions or specify its apparent migration. Verify the higher band before assigning it to a TXNRD1 dimer.
Consider the seven annotated isoforms and TXNRD1 homodimerization as possibilities, then check whether each band's detection fits the antibody epitope. The listed modifications at UniProt positions 218 and 281 do not, by themselves, identify an unexpected band. Confirm band identity experimentally.
TXNRD1 is annotated as a homodimer, making dimerization relevant when evaluating a higher band. The annotation does not show that a dimer survives your Western blot conditions or specify its apparent migration. Verify the higher band before assigning it to a TXNRD1 dimer.
Consider the seven annotated isoforms and TXNRD1 homodimerization as possibilities, then check whether each band's detection fits the antibody epitope. The listed modifications at UniProt positions 218 and 281 do not, by themselves, identify an unexpected band. Confirm band identity experimentally.
Boster reagents
TXNRD1 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-TXNRD1 Antibody Picoband®
Cat # A01778-3
Real WB data
Anti-TXNRD1 Rabbit Monoclonal Antibody
Cat # M01778
Both listed anti-TXNRD1 antibodies have WB images. A01778-3 shows an approximately 65 kDa band in human cell lysates; M01778 shows an approximately 55 kDa band in human, rat, and mouse samples. Both bands differ from the stated expected size of 71 kDa.
Which to pick: For human samples, consider the different observed band sizes when choosing between A01778-3 and M01778. For mouse or rat samples, M01778 is the listed option with WB images from those species. Confirm the band in your own sample.
Source: BosterBio TXNRD1 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q16881.
- Human Protein Atlas. TXNRD1 tissue expression.
- PMC4694848 — target-verified WB comparison
- PMC13556941 — target-verified WB comparison
- PMC5425838 — target-verified WB comparison