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- Table of Contents
CHD1 / ATP-dependent chromatin remodeler CHD1 · Western blot design guide
Design a Western Blot for CHD1
Real validated CHD1 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-CHD1 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
CHD1 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 | ~196.7 kDa | |
| Observed band | ~270 kDa | |
| Gel | 5–20% (catalog A01175-1) | |
| Positive control | Appendix (IHC candidate; verify WB) +4 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 C7 membership | |
| Isoform | 2 isoform(s) |
Section 1
Real Curated CHD1 Western Blot Protocols
The A01175-1 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human Hela, human 293T, human K562 (catalog A01175-1) |
| Gel % | 5–20% (catalog A01175-1) |
| Load | 30 ug; reducing conditions (catalog A01175-1) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog A01175-1) |
| Membrane | nitrocellulose membrane (catalog A01175-1) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog A01175-1) |
| Primary antibody | A01175-1 · 0.5 μg/mL (catalog A01175-1) |
| Primary incubation | overnight at 4°C (catalog A01175-1) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog A01175-1) |
| Secondary incubation | 1.5 hour at RT (catalog A01175-1) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog A01175-1) |
| Detection | ECL (catalog A01175-1) |
Section 2
What Is the Expected CHD1 Western Blot Band Size?
CHD1 is predicted at 196.7 kDa but observed near 270 kDa on a reducing blot; the cause of the difference is not established.
| Band near 270 kDa | Empirical CHD1 band in reducing whole-cell lysates; confirm identity with controls |
| Band near 196.7 kDa | Near the UniProt predicted mass; identity requires confirmation |
| Multiple discrete bands | Isoforms 1 and 2 are documented, but distinct migration is unproven |
| Weak band in a cytoplasmic fraction | CHD1 is nuclear and chromatin-associated outside mitosis |
💡Expected CHD1 appearanceUniProt predicts 196.7 kDa, while antibody QC detects CHD1 near 270 kDa in reducing whole-cell lysates; the difference is unexplained, so confirm band identity with appropriate controls.
| UniProt predicted mass | 196.7 kDa calculated; the observed band is near 270 kDa for an unestablished reason |
| Isoform 1 | Its individual mass and migration are not supplied |
| Isoform 2 | Its size relative to isoform 1 is not supplied |
| Documented phosphorylation | Phosphoserine and phosphothreonine sites are listed, but no apparent size effect is established |
| Situation | Likely cause | Next action |
|---|---|---|
| Band higher than expected | The observed CHD1 band is near 270 kDa despite a 196.7 kDa predicted mass; the cause is unknown | Compare with the reported reducing blot and confirm identity by CHD1 depletion |
| Band lower than expected | An additional band may represent an isoform or an unrelated signal; isoform masses are unavailable | Check whether it decreases after CHD1 depletion |
| Multiple bands | Two isoforms are documented, but their distinct migration is unproven | Use CHD1 depletion and an independent antibody to assess band identity |
| Weak or no signal | CHD1 is associated with nuclear chromatin outside mitosis | Check nuclear extraction and include a positive whole-cell lysate control |
| Fragments below expected size | Sample degradation or nonspecific antibody binding; no cleavage feature is supplied | Process samples promptly and test whether the bands decrease after CHD1 depletion |
Experiment evidence
Western blot images showing CHD1 band
A01175-1 · CHD1
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for CHD1 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for CHD1 in Western blot, you can use appendix tissue lysate.
Positive control: Appendix (IHC candidate; verify WB)
Negative control: Adipose tissue (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain such as Ponceau alongside the samples.
⚠️Feasibility: CHD1 moves from the nucleus to the cytoplasm during mitosis, so fractionation and cell-cycle state may affect signal.
HPA tissue expression evidence for CHD1
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 |
|---|---|---|---|---|
| Appendix | glandular cells | High | Protein (IHC) | HPA → |
| Colon | glandular cells | High | Protein (IHC) | HPA → |
| Kidney | cells in tubules | High | Protein (IHC) | HPA → |
| Ovary | follicle cells | High | Protein (IHC) | HPA → |
| Placenta | decidual cells | High | 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 → |
| Bone marrow | hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Lung | alveolar cells | Not detected | Protein (IHC) | HPA → |
| Seminal vesicle | glandular cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced CHD1 Western Blot Tips
Deeper troubleshooting and optimisation questions for CHD1, answered from its protein features.
How should CHD1 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 CHD1 isoforms produce separate bands?
Isoforms · UniProt lists isoforms 1 and 2 and a sequence difference at canonical position 1684, which is missing in isoform 2. This small listed difference alone does not establish resolvable bands. Check which isoform your antibody recognizes before assigning bands.
Which CHD1 modifications matter when interpreting bands?
PTM · UniProt lists 26 modified residues, including phosphoserines 215, 216, 1096–1102, 1353–1373, and 1689, plus phosphothreonines 237 and 250. These are UniProt canonical coordinates; antibody or paper numbering may differ. Phosphorylation may affect migration, but the listed sites alone do not establish a visible shift.
Does this guide establish induction of CHD1?
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 CHD1 Western blot?
Transfer · CHD1 has a predicted mass of 196.7 kDa. Use a transfer setup suitable for a large protein, and verify transfer near 200 kDa with a marker or membrane stain before interpreting a weak or absent band.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A01175-1 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 CHD1 be quantified across cell-cycle samples?
Quantitation · UniProt notes that CHD1 moves from chromatin to the cytoplasm during mitosis and returns during telophase-cytokinesis. For comparisons across cell-cycle stages, keep the sampled fraction consistent and interpret changes in a nuclear or chromatin fraction alongside this redistribution.
Why might CHD1 appear near 270 kDa instead of 196.7 kDa?
Interpretation · UniProt lists a predicted mass of 196.7 kDa and multiple phosphorylation sites, but these features do not establish why an observed band appears near 270 kDa. Check that the band is reproducible and verify its identity with a second CHD1 antibody or CHD1 depletion.
UniProt lists two isoforms, multiple phosphorylation sites, and a predicted mass of 196.7 kDa. None alone identifies an unexpected band. Compare its detection with a second CHD1 antibody or CHD1 depletion before assigning it to an isoform or modification.
Yes. UniProt places CHD1 in the nucleus and cytoplasm and notes its release into the cytoplasm during mitosis. Record the cell-cycle stage and compare like fractions when interpreting signal differences.
UniProt lists two isoforms, multiple phosphorylation sites, and a predicted mass of 196.7 kDa. None alone identifies an unexpected band. Compare its detection with a second CHD1 antibody or CHD1 depletion before assigning it to an isoform or modification.
Yes. UniProt places CHD1 in the nucleus and cytoplasm and notes its release into the cytoplasm during mitosis. Record the cell-cycle stage and compare like fractions when interpreting signal differences.
Boster reagents
CHD1 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-CHD1 Antibody Picoband®
Cat # A01175-1
The catalog reports one anti-CHD1 antibody, A01175-1, for Western blotting. Its image shows a band near 270 kDa in six human cell lysates, although the stated expected size is 196 kDa. This discrepancy limits interpretation of the band.
Which to pick: A01175-1 is the only listed option. It reports human, mouse, and rat reactivity, but the supplied WB image uses human cell lysates only. Consider the 270 kDa observed band versus the 196 kDa expected size when planning interpretation.
Source: BosterBio CHD1 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry O14646.
- Human Protein Atlas. CHD1 tissue expression.
- PMC4946808 — target-verified WB comparison
- PMC11042865 — target-verified WB comparison
- PMC9444962 — target-verified WB comparison