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- Table of Contents
NCOA1 / Nuclear receptor coactivator 1 · Western blot design guide
Design a Western Blot for NCOA1
Real validated NCOA1 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-NCOA1 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
NCOA1 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 | ~156.8 kDa | |
| Observed band | ~157 kDa | |
| Gel | 8% (catalog M00856) | |
| Positive control | Adrenal gland (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 + Acetylated | |
| Caveat | Phosphorylation-state controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 3 isoform(s) |
Section 1
Real Curated NCOA1 Western Blot Protocols
The M00856 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human 293T, human HEL, human K562 (catalog M00856) |
| Gel % | 8% (catalog M00856) |
| Load | 30 ug; reducing conditions (catalog M00856) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog M00856) |
| Membrane | nitrocellulose membrane (catalog M00856) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog M00856) |
| Primary antibody | M00856 · 1: 500 (catalog M00856) |
| Primary incubation | overnight at 4°C (catalog M00856) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog M00856) |
| Secondary incubation | 1.5 hour at RT (catalog M00856) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog M00856) |
| Detection | ECL (catalog M00856) |
Section 2
What Is the Expected NCOA1 Western Blot Band Size?
NCOA1 is predicted at 156.8 kDa and observed near 157 kDa; the evidence does not establish a cause for the small difference.
| Band near 157 kDa | Consistent with the observed NCOA1 band; confirm identity with antibody controls |
| Several bands at different positions | NCOA1 isoforms 1, 2, and 3 could contribute; their migration is not established |
| Band shifted from 157 kDa | Phosphorylation could affect mobility; the shift and band identity need verification |
| Faint band in whole-cell lysate, stronger in nuclear extract | Consistent with NCOA1 nuclear localization |
💡Expected NCOA1 appearanceNCOA1 has a predicted mass of 156.8 kDa and an observed band near 157 kDa in reducing whole-cell blots; confirm band identity with appropriate antibody controls.
| UniProt predicted mass | Places the full-length reference near 156.8 kDa; the observed band is near 157 kDa |
| Splice isoform 1 | Its individual mass and migration are not supplied |
| Splice isoform 2 | Could differ in size from other isoforms; its mass and migration are not supplied |
| Splice isoform 3 | Could differ in size from other isoforms; its mass and migration are not supplied |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Nuclear NCOA1 may be poorly recovered during extraction | Check nuclear extraction and a nuclear marker |
| Band higher than expected | Identity and cause of altered migration are unestablished | Compare with the 157 kDa reference and verify with an independent NCOA1 antibody |
| Band lower than expected | An isoform or nonspecific band is possible; individual isoform masses are unknown | Check antibody specificity and compare with the 157 kDa band |
| Multiple bands | NCOA1 has three named splice isoforms, but distinct band positions are unconfirmed | Verify candidate bands with an independent antibody or NCOA1 depletion |
| Weak or no signal | Nuclear protein may be underrepresented in the loaded extract | Assess loading and compare whole-cell with nuclear extract |
| Fragments below expected size | Protein degradation is possible; fragment masses are not supplied | Repeat with fresh, protease-inhibited extract and verify band identity |
Experiment evidence
Western blot images showing NCOA1 band
M00856 · NCOA1
RP1056 · NCOA1
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for NCOA1 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for NCOA1 in Western blot, you can use adrenal gland tissue, which HPA scores as high for NCOA1.
Positive control: Adrenal gland (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: HPA provides a tissue negative control, while nuclear enrichment may help detect nuclear NCOA1.
HPA tissue expression evidence for NCOA1
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 → |
| Breast | glandular cells | High | Protein (IHC) | HPA → |
| Bronchus | basal cells | High | Protein (IHC) | HPA → |
| Cerebellum | cells in granular layer | High | Protein (IHC) | HPA → |
| Cerebral cortex | glial 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 → |
| Heart muscle | cardiomyocytes | Low | Protein (IHC) | HPA → |
| Liver | hepatocytes | Low | Protein (IHC) | HPA → |
| Skeletal muscle | myocytes | Low | Protein (IHC) | HPA → |
| Soft tissue | fibroblasts | Low | Protein (IHC) | HPA → |
Section 3
Advanced NCOA1 Western Blot Tips
Deeper troubleshooting and optimisation questions for NCOA1, answered from its protein features.
How should NCOA1 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 NCOA1 isoforms produce different bands?
Isoforms · UniProt lists isoforms 1, 2, and 3. In UniProt coordinates, isoform 3 lacks residue 1385, while isoform 2 replaces residues 1386–1441 with a shorter sequence. These changes could affect band position, but the features do not establish which isoform produces a given band.
Which NCOA1 phosphorylation sites matter when interpreting bands?
PTM · Listed sites in UniProt coordinates are Ser22, Ser372, Ser395, Ser517, Ser558, Ser569, Ser698, Ser1033, Thr1179, Ser1185, and Ser1372. A phosphatase comparison can test whether phosphorylation contributes to band differences; site presence alone does not establish a visible shift. Check numbering conventions before comparing antibody or paper site labels.
UniProt lists N-acetylserine at residue 2 and asymmetric dimethylarginine at residues 1073, 1091, 1124, and 1131, using its sequence coordinates. These features identify possible modified forms but do not establish separate visible bands.
UniProt lists N-acetylserine at residue 2 and asymmetric dimethylarginine at residues 1073, 1091, 1124, and 1131, using its sequence coordinates. These features identify possible modified forms but do not establish separate visible bands.
Does this guide establish induction of NCOA1?
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 NCOA1 Western blot?
Transfer · NCOA1 is predicted at 156.8 kDa, so use transfer conditions suitable for a large protein. Check the membrane around 157 kDa and inspect the gel after transfer to see whether protein remains. The supplied features do not specify a unique transfer method.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the M00856 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 NCOA1 abundance be quantified?
Quantitation · NCOA1 is listed as nuclear. For comparisons, use the same sample fraction across conditions and normalize the NCOA1 signal to a suitable loading measure from that fraction. If using nuclear extracts, compare their loading consistently.
Does a band near 157 kDa match predicted NCOA1 mass?
Interpretation · Yes. The observed band near 157 kDa closely matches the predicted 156.8 kDa. The listed modifications alone do not establish a visible shift or explain a mass difference.
Compare bands with the expected ~157 kDa position and consider the documented isoforms and modifications. Isoform 2 has a shorter replacement at UniProt residues 1386–1441; isoform 3 lacks residue 1385. The features cannot assign an unexpected band to an isoform or modification on position alone.
Compare bands with the expected ~157 kDa position and consider the documented isoforms and modifications. Isoform 2 has a shorter replacement at UniProt residues 1386–1441; isoform 3 lacks residue 1385. The features cannot assign an unexpected band to an isoform or modification on position alone.
Boster reagents
NCOA1 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-KAT13A / SRC1 Monoclonal Antibody
Cat # M00856
Real WB data
Anti-KAT13A/SRC1/NCOA1 Antibody Picoband®
Cat # RP1056
Both listed anti-NCOA1 antibodies have Western blot images reporting a band near the expected 157 kDa. M00856 was shown with human cell lysates; RP1056 was shown with human and rodent cell or tissue lysates. The supplied evidence does not establish specificity by independent validation.
Which to pick: For mouse or rat samples, consider RP1056, whose image includes mouse and rat lysates. For human samples, both have WB images: M00856 shows 293T, HEL, K562 and RT4; RP1056 shows MCF-7, PC-3, 293T and HeLa. Match your sample to the reported contexts.
Source: BosterBio NCOA1 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q15788.
- Human Protein Atlas. NCOA1 tissue expression.
- PMC12821625 — target-verified WB comparison