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CHD8 / ATP-dependent chromatin remodeler CHD8 · Western blot design guide
Design a Western Blot for CHD8
Source-linked CHD8 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-CHD8 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
CHD8 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 | ~290.5 kDa | |
| Observed band | ~300 kDa | |
| Gel | 5–20% (catalog A01346-1) | |
| Negative control | Suggested KO / knockdown lysate |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Phosphorylated + Ubl conjugation | |
| Caveat | Modification-state controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 2 isoform(s) |
Section 1
Source-Linked CHD8 Western Blot Protocol Options
The A01346-1 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human HepG2 (catalog A01346-1) |
| Gel % | 5–20% (catalog A01346-1) |
| Load | 30 ug; reducing conditions (catalog A01346-1) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog A01346-1) |
| Membrane | nitrocellulose membrane (catalog A01346-1) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog A01346-1) |
| Primary antibody | A01346-1 · 0.5 μg/mL (catalog A01346-1) |
| Primary incubation | overnight at 4°C (catalog A01346-1) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog A01346-1) |
| Secondary incubation | 1.5 hour at RT (catalog A01346-1) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog A01346-1) |
| Detection | ECL (catalog A01346-1) |
Section 2
What Is the Expected CHD8 Western Blot Band Size?
CHD8 is predicted at 290.5 kDa and observed near 300 kDa; the cause of the difference is not established.
| Band near 300 kDa | Empirical CHD8 band in a reducing whole-cell lysate blot |
| Band near 290.5 kDa | Consistent with the predicted CHD8 mass; confirm identity with controls |
| Multiple high-mass bands | CHD8 isoforms 1 and 2 are possible contributors, but distinct migration is unproven |
| Weak or absent band in lysate | Nuclear CHD8 may be poorly recovered during extraction |
💡Expected CHD8 appearanceCHD8 has a predicted mass of 290.5 kDa and an empirical band near 300 kDa; the cause of the difference is unestablished, so confirm band identity with appropriate controls.
| UniProt predicted mass | 290.5 kDa is the sequence-based reference; a band near 300 kDa was observed |
| Splice isoform 1 | May differ in size from isoform 2; its individual mass and migration are unspecified |
| Splice isoform 2 | May differ in size from isoform 1; its individual mass and migration are unspecified |
| Alternative splicing of isoforms 1 and 2 | Could affect apparent size, but distinct resolvable bands are unestablished |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Nuclear CHD8 may be poorly extracted | Check nuclear extraction and probe a nuclear fraction |
| Band higher than expected | The observed CHD8 band is near 300 kDa versus a predicted 290.5 kDa; the cause is unestablished | Check the high-mass marker range and confirm band identity with a CHD8 control |
| Band lower than expected | Isoforms 1 and 2 exist, but their individual masses are unspecified | Confirm the lower band's identity with an independent CHD8 antibody or depletion control |
| Multiple bands | CHD8 has two splice isoforms; their separation on this blot is unestablished | Compare bands using an independent CHD8 antibody or depletion control |
| Weak or no signal | Recovery of nuclear CHD8 may be low | Check nuclear protein recovery and transfer of high-mass proteins |
Experiment evidence
Western blot images showing CHD8 band
A01346-1 · CHD8
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for CHD8 Western blot
🧪For positive controls for CHD8 in Western blot, you can use a confirmed CHD8-positive nuclear lysate; the supplied HPA data identify no specific tissue or cell line.
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 provides no expression data here, so a positive sample needs independent validation and no negative tissue can be identified.
HPA tissue expression evidence for CHD8
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 CHD8 Western Blot Tips
Deeper troubleshooting and optimisation questions for CHD8, answered from its protein features.
How should CHD8 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 CHD8 isoforms affect the bands detected?
Isoforms · UniProt lists two isoforms. In isoform 2, residues 1–281 of the canonical sequence are replaced by MK, so it may migrate differently. Check whether the antibody epitope lies in that N-terminal region when interpreting bands.
Which CHD8 phosphorylation sites matter when interpreting a blot?
PTM · In canonical UniProt numbering, phosphoserines occur at 432, 553, 562, 1420, 1424, 1976, 1978, 1995, 2008, 2046, 2069, 2071, 2182, 2200, 2202, 2211, 2223 and 2519; phosphothreonines occur at 1993, 2051, 2204 and 2215. Check the numbering convention of any phospho-specific antibody before comparing sites.
CHD8 has 22 listed phosphorylated residues, but their presence does not demonstrate a visible band shift or explain the roughly 300 kDa apparent band. Treat a shifted band as unresolved until its identity is verified.
CHD8 has 22 listed phosphorylated residues, but their presence does not demonstrate a visible band shift or explain the roughly 300 kDa apparent band. Treat a shifted band as unresolved until its identity is verified.
Does this guide establish induction of CHD8?
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 CHD8 Western blot?
Transfer · CHD8 is a large protein, with a predicted mass of 290.5 kDa. Choose transfer conditions suitable for proteins around 300 kDa and check transfer efficiency in that region. The supplied features do not specify a particular transfer method.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A01346-1 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 CHD8 abundance be quantified?
Quantitation · CHD8 is annotated as nuclear. For comparisons across samples, quantify the same CHD8 band with consistent loading and an appropriate normalization signal. If using nuclear fractions, keep fraction preparation consistent; the features do not establish that every CHD8-reactive band represents full-length protein.
Why might CHD8 appear near 300 kDa rather than 290.5 kDa?
Interpretation · The supplied apparent band is about 300 kDa, versus a predicted mass of 290.5 kDa. Compare it with a high molecular weight marker. The listed features alone do not establish why the apparent and predicted masses differ.
First compare bands with the roughly 300 kDa observed band and consider the shorter N-terminus of isoform 2. Check antibody epitope coverage and whether a band is enriched in nuclear material. The listed phosphorylation sites alone do not identify an unexpected band or prove a mobility shift.
First compare bands with the roughly 300 kDa observed band and consider the shorter N-terminus of isoform 2. Check antibody epitope coverage and whether a band is enriched in nuclear material. The listed phosphorylation sites alone do not identify an unexpected band or prove a mobility shift.
Boster reagents
CHD8 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-CHD8 Antibody Picoband®
Cat # A01346-1
The catalog reports one anti-CHD8 antibody, A01346-1, with stated human reactivity and a Western blot image using human HepG2 whole-cell lysate. The caption reports a band near 300 kDa, matching the expected size. Evidence supplied is limited to this sample.
Which to pick: A01346-1 is the only listed option. Its WB image documents human HepG2 lysate under reducing conditions, with 30 μg loaded and primary antibody at 0.5 μg/mL. Choose it when those documented conditions fit your experiment; other sample types are not shown.
Source: BosterBio CHD8 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q9HCK8.
- Human Protein Atlas. CHD8 tissue expression.
- PMC8536677 — target-verified WB comparison
- PMC7035710 — target-verified WB comparison
- PMC12106843 — target-verified WB comparison