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- Table of Contents
ATRX / Transcriptional regulator ATRX · Western blot design guide
Design a Western Blot for ATRX
Source-linked ATRX Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-ATRX 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
ATRX 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 | ~282.6 kDa | |
| Observed band | 300 kDa | |
| Gel | 8% (catalog A00203-2) | |
| Positive control | Adipose tissue (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 | 6 isoform(s) |
Section 1
Source-Linked ATRX Western Blot Protocol Options
The A00203-2 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human A549, human SH-SY5Y, human MOLT4, rat C6, mouse NIH/3T3 (catalog A00203-2) |
| Gel % | 8% (catalog A00203-2) |
| Load | 30 ug; reducing conditions (catalog A00203-2) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog A00203-2) |
| Membrane | nitrocellulose membrane (catalog A00203-2) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog A00203-2) |
| Primary antibody | A00203-2 · 0.5 μg/mL (catalog A00203-2) |
| Primary incubation | overnight at 4°C (catalog A00203-2) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog A00203-2) |
| Secondary incubation | 1.5 hour at RT (catalog A00203-2) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog A00203-2) |
| Detection | ECL (catalog A00203-2) |
Section 2
What Is the Expected ATRX Western Blot Band Size?
ATRX is predicted at 282.6 kDa and observed near 300 kDa; the cause of the difference is not established.
| Band near 300 kDa | matches the observed ATRX band; confirm identity with antibody controls |
| Band near 283 kDa | aligns with the sequence-predicted mass; identity requires confirmation |
| Additional bands at different sizes | could reflect ATRX isoforms, but distinct migration has not been demonstrated |
| Weak whole-cell band with stronger nuclear signal | is consistent with ATRX nuclear localization |
💡Expected ATRX appearanceATRX has a predicted mass of 282.6 kDa and an observed band near 300 kDa; the cause of the difference is unestablished, so confirm band identity with antibody controls.
| UniProt predicted mass | 282.6 kDa is the sequence-based reference; the observed band is near 300 kDa |
| Isoforms 1 and 2 | sequence differences may affect migration; their individual band sizes are unknown |
| Isoforms 3 and 4 | sequence differences may affect migration; their individual band sizes are unknown |
| Isoforms 5 and 6 | sequence differences may affect migration; their individual band sizes are unknown |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | nuclear ATRX may be poorly recovered | check nuclear extraction and a known positive lysate |
| Band higher than expected | the observed ATRX band is near 300 kDa, above its 282.6 kDa predicted mass; the cause is unestablished | compare with the antibody QC band and confirm identity with an independent antibody |
| Band lower than expected | an isoform or degraded ATRX is possible, but neither is established for the band | check sample integrity and confirm identity with an independent antibody |
| Multiple bands | ATRX has six isoforms, though distinct bands are unproven | compare antibody epitopes and confirm bands with an independent antibody |
| Weak or no signal | nuclear ATRX recovery may be low | check nuclear fraction recovery and include a known positive lysate |
Experiment evidence
Western blot images showing ATRX band
A00203-2 · ATRX
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for ATRX Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for ATRX in Western blot, you can use adipose tissue lysate.
Positive control: Adipose tissue (IHC candidate; verify WB)
Negative control: Suggested KO / knockdown lysate
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside.
⚠️Feasibility: No tissue is reported as not detected, so use siRNA knockdown or a KO line for a negative control.
HPA tissue expression evidence for ATRX
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 |
|---|---|---|---|---|
| Adipose tissue | adipocytes | High | Protein (IHC) | HPA → |
| Adrenal gland | glandular cells | High | Protein (IHC) | HPA → |
| Appendix | endocrine cells | High | Protein (IHC) | HPA → |
| Bone marrow | hematopoietic cells | High | Protein (IHC) | HPA → |
| Breast | adipocytes | High | Protein (IHC) | HPA → |
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Liver | cholangiocytes | Low | Protein (IHC) | HPA → |
| Parathyroid gland | glandular cells | Low | Protein (IHC) | HPA → |
| Cervix | glandular cells | Medium | Protein (IHC) | HPA → |
| Endometrium | cells in endometrial stroma | Medium | Protein (IHC) | HPA → |
| Lymph node | non-germinal center cells | Medium | Protein (IHC) | HPA → |
Section 3
Advanced ATRX Western Blot Tips
Deeper troubleshooting and optimisation questions for ATRX, answered from its protein features.
How should ATRX 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.
Which ATRX isoforms could produce smaller bands?
Isoforms · Six isoforms are listed. Relative to the supplied sequence, isoform 1 lacks residues 1–204; isoforms 2 and 5 lack 1–117; and isoform 6 lacks 124–162, 573–601, and 1419–2492. Isoforms 3 and 5 also lack 124–161. Isoform 6's large C-terminal deletion makes a substantially smaller product plausible, but band size alone cannot identify an isoform.
Could ATRX phosphorylation affect its band pattern?
PTM · UniProt lists phosphoserine, phosphothreonine, and phosphotyrosine sites, including clustered sites at 1011–1013 and 1322, 1324, and 1326. These are coordinates in the supplied UniProt sequence; other numbering conventions may differ. Their presence does not establish a visible shift. Compare samples under the same preparation and gel conditions before interpreting pattern changes.
The supplied features list N6-acetyllysine at UniProt position 967 and omega-N-methylarginine at positions 2474 and 2480. These coordinates refer to the supplied sequence. Isoform 6 lacks residues 1419–2492, including both listed methylarginine positions. None of these annotations alone predicts a separate visible band.
The supplied features list N6-acetyllysine at UniProt position 967 and omega-N-methylarginine at positions 2474 and 2480. These coordinates refer to the supplied sequence. Isoform 6 lacks residues 1419–2492, including both listed methylarginine positions. None of these annotations alone predicts a separate visible band.
Does this guide establish induction of ATRX?
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 ATRX?
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 A00203-2 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 ATRX bands be quantified consistently?
Quantitation · ATRX is listed in the nucleus, including chromosomes and PML bodies. Compare like samples and use the same band region near the observed 300 kDa position across lanes. If additional bands appear, report them separately rather than combining their intensities with the main band without evidence that they represent ATRX.
Why might ATRX run near 300 kDa instead of 282.6 kDa?
Interpretation · The observed band is approximately 300 kDa, while the supplied predicted mass is 282.6 kDa. ATRX has multiple modified residues and isoforms, but these features alone do not establish the cause of the apparent mass difference. Use the 300 kDa band as an empirical reference, not proof of a particular modification.
Check whether its size is compatible with the listed sequence deletions, especially isoform 6's missing residues 1419–2492. Other isoforms have shorter deletions. The supplied features do not identify any particular unexpected band, so apparent size alone cannot distinguish an isoform from another explanation.
Check whether its size is compatible with the listed sequence deletions, especially isoform 6's missing residues 1419–2492. Other isoforms have shorter deletions. The supplied features do not identify any particular unexpected band, so apparent size alone cannot distinguish an isoform from another explanation.
Boster reagents
ATRX 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-ATRX Antibody Picoband®
Cat # A00203-2
The catalog reports A00203-2 for ATRX Western blotting, with a blot image showing a reported band near 300 kDa versus the expected 283 kDa. The caption documents human, rat, and mouse cell lysates; evidence is limited to the supplied product image.
Which to pick: A00203-2 is the only listed option. Its stated reactivity covers human, mouse, and rat, and its WB image includes A549, SH-SY5Y, MOLT4, C6, and NIH/3T3 lysates. Choose it when those reported contexts fit your experiment.
Source: BosterBio ATRX gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P46100.
- Human Protein Atlas. ATRX tissue expression.
- PMC12727880 — target-verified WB comparison
- PMC12900699 — target-verified WB comparison
- PMC5662737 — target-verified WB comparison