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- Table of Contents
CREBBP / CREB-binding protein · Western blot design guide
Design a Western Blot for CREBBP
Real validated CREBBP Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-CREBBP 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
CREBBP 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 | ~265.4 kDa | |
| Gel | 4–12% gradient (standard starting point) | |
| Positive control | Cerebellum (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 | Phosphorylation-state controls | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 2 isoform(s) |
Section 1
Real Curated CREBBP Western Blot Protocols
The A00205-1 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | various cells (catalog A00205-1) |
| Gel % | 4–12% gradient (standard starting point) |
| Load | 20–30 µg total protein per lane; optimize for abundance (standard starting point) |
| Transfer | Wet/tank transfer; optimize duration (standard starting point) |
| Membrane | 0.45 µm PVDF (standard starting point) |
| Blocking | 5% milk or 5% BSA in TBST (standard starting point) |
| Primary antibody | A00205-1 · 1:1000 (catalog A00205-1) |
| 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 CREBBP Western Blot Band Size?
CREBBP has a predicted full-length mass of 265.4 kDa; isoforms and modifications are documented, but their effects on band migration are not demonstrated.
| Band near 265.4 kDa | Consistent with the predicted full-length CREBBP mass; confirm its identity. |
| Additional band at another position | Could represent isoform 1 or 2, but their migration is not established. |
| Band in a nuclear fraction | Consistent with CREBBP's reported nuclear location. |
| Band in a cytoplasmic fraction | Consistent with CREBBP's reported cytoplasmic location. |
💡Expected CREBBP appearanceUniProt predicts 265.4 kDa for full-length CREBBP; no empirical band size is supplied, and the listed isoforms and modifications do not establish a particular migration pattern.
| UniProt predicted full-length mass | Sets a calculated reference of 265.4 kDa, not a measured band position. |
| Full-length 2442-residue sequence | Underlies the calculated mass without establishing apparent migration. |
| Isoform 1 | May differ in size from isoform 2; no isoform-specific mass is supplied. |
| Isoform 2 | May differ in size from isoform 1; no isoform-specific mass is supplied. |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | CREBBP may be easier to detect in a nuclear-enriched fraction. | Check a nuclear fraction and a positive-control lysate. |
| Band higher than expected | Apparent migration may differ from calculated mass; the cause is unestablished. | Check peptide blocking and an independent CREBBP antibody. |
| Band lower than expected | An alternative isoform is possible, but its mass is unknown. | Confirm identity with peptide blocking or an independent antibody. |
| Multiple bands | Isoforms 1 and 2 are listed, but distinct bands are unverified. | Compare bands with peptide blocking or an independent antibody. |
| Weak or no signal | CREBBP distribution between nucleus and cytoplasm may affect sample signal. | Check nuclear and cytoplasmic fractions with appropriate fraction controls. |
Experiment evidence
Western blot images showing CREBBP band
A00205-1 · CREBBP
A00205-1 · CREBBP
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for CREBBP Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for CREBBP in Western blot, you can use cerebellum lysate, which has high HPA expression.
Positive control: Cerebellum (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 CREBBP
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 |
|---|---|---|---|---|
| Cerebellum | cells in granular layer | High | Protein (IHC) | HPA → |
| Cervix | squamous epithelial cells | High | Protein (IHC) | HPA → |
| Colon | glandular cells | High | Protein (IHC) | HPA → |
| Endometrium | cells in endometrial stroma | High | Protein (IHC) | HPA → |
| Esophagus | squamous epithelial 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 → |
| Bone marrow | hematopoietic cells | Low | Protein (IHC) | HPA → |
| Hippocampus | glial cells | Low | Protein (IHC) | HPA → |
| Skeletal muscle | myocytes | Low | Protein (IHC) | HPA → |
| Adrenal gland | glandular cells | Medium | Protein (IHC) | HPA → |
Section 3
Advanced CREBBP Western Blot Tips
Deeper troubleshooting and optimisation questions for CREBBP, answered from its protein features.
Where should the main CREBBP band appear?
Band shift · The canonical CREBBP sequence is 2,442 residues with a predicted mass of 265.4 kDa. Use that as a reference when identifying the band; no empirical apparent mass is supplied, and the listed features do not establish a visible shift.
Could CREBBP isoforms produce different bands?
Isoforms · UniProt lists two isoforms. In isoform 2, residues 406–444 of the canonical sequence are replaced by one alanine. This sequence difference could affect band position, but the supplied features do not show whether the isoforms resolve on a blot.
Which CREBBP modifications matter when interpreting band shifts?
PTM · UniProt lists phosphorylation, acetylation, and arginine methylation, including a cluster of acetylated lysines at positions 1583–1597. These sites identify possible sources of molecular variation, but their presence alone does not demonstrate a visible shift or explain any observed mass difference. Coordinates here use the supplied UniProt sequence numbering.
UniProt assigns phosphoserine 124 to ATM and phosphoserines 1382 and 1386 to IKKA. For a phosphorylation comparison, probe the relevant sites alongside total CREBBP and compare matched samples. These assignments do not establish that a particular treatment increases phosphorylation or changes band position.
UniProt assigns phosphoserine 124 to ATM and phosphoserines 1382 and 1386 to IKKA. For a phosphorylation comparison, probe the relevant sites alongside total CREBBP and compare matched samples. These assignments do not establish that a particular treatment increases phosphorylation or changes band position.
Does this guide establish induction of CREBBP?
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 CREBBP Western blot?
Transfer · CREBBP's predicted mass is 265.4 kDa, so use a transfer setup suitable for a protein of that size and check transfer near 265 kDa with a total-protein stain. The supplied features do not specify a membrane, buffer, or transfer time.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A00205-1 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 CREBBP abundance be quantified across fractions?
Quantitation · UniProt lists CREBBP in both the nucleus and cytoplasm and notes recruitment to nuclear bodies by SS18L1/CREST. Quantify the same sample fraction across conditions and report which fraction was measured; a change in one fraction need not represent a change in total cellular CREBBP.
How should unexpected CREBBP bands be interpreted?
Interpretation · Compare them with the 265.4 kDa prediction and consider the documented isoform 2 sequence change and modified residues. The supplied record lists no signal peptide, propeptide, or glycosylation sites. Band position alone cannot assign an unexpected band to an isoform or modification.
Boster reagents
CREBBP 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-CBP CREBBP Antibody
Cat # A00205-1
The listed anti-CREBBP antibody is a polyclonal reagent with a Western blot image described as analysis of various cells at 1:1000 dilution. The supplied caption does not identify the cells or demonstrate validation in each listed species.
Which to pick: A00205-1 is the only listed option. It reports Human, Mouse, and Rat reactivity and includes a Western blot image; check suitability for your sample, since the caption identifies the samples only as various cells.
Source: BosterBio CREBBP gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q92793.
- Human Protein Atlas. CREBBP tissue expression.
- PMC8210551 — target-verified WB comparison
- PMC5386396 — target-verified WB comparison
- PMC7362711 — target-verified WB comparison