This website uses cookies to ensure you get the best experience on our website.
- Table of Contents
MACC1 / Metastasis-associated in colon cancer protein 1 · Western blot design guide
Design a Western Blot for MACC1
Source-linked MACC1 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-MACC1 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
MACC1 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 | ~96.6 kDa | |
| Gel | 8–10% (standard starting point) | |
| 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 | |
| Caveat | Phosphorylation-state controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 1 isoform(s) |
Section 1
Source-Linked MACC1 Western Blot Protocol Options
The A04732 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | mouse liver tissue lysate (catalog A04732) |
| Gel % | 8–10% (standard starting point) |
| Load | 20–30 µg total protein per lane; optimize for abundance (standard starting point) |
| Transfer | Standard semi-dry transfer; verify efficiency (standard starting point) |
| Membrane | 0.45 µm PVDF (standard starting point) |
| Blocking | 5% milk or 5% BSA in TBST (standard starting point) |
| Primary antibody | A04732 · (A) 1 and (B) 2 μg/mL (catalog A04732) |
| 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 MACC1 Western Blot Band Size?
MACC1 has a predicted mass of 96.6 kDa; Ser19 phosphorylation is annotated, but no migration effect or empirical band size is demonstrated.
| Single band near 96.6 kDa | consistent with the predicted full-length MACC1 mass; confirm identity with a specificity control |
| Band near 96.6 kDa in cytoplasmic fraction | consistent with MACC1's cytoplasmic localization |
| Band near 96.6 kDa in nuclear fraction | consistent with MACC1's nuclear localization |
| Stronger cytoplasmic than nuclear band | consistent with MACC1 being mainly cytoplasmic in non-metastasizing tumors |
💡Expected MACC1 appearanceUniProt predicts 96.6 kDa for MACC1; no empirical band size is supplied, so confirm a candidate band with antibody specificity controls.
| Predicted molecular mass | sets a 96.6 kDa reference for full-length MACC1 |
| 852-residue sequence | underlies the predicted full-length mass; no cleavage is annotated |
| Phosphoserine at Ser19 | has no demonstrated effect on apparent band size in the supplied evidence |
| Single annotated isoform | provides no evidence for isoform-dependent band sizes |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | MACC1 signal may be below detection in the sampled lysate | check antibody performance and compare cytoplasmic and nuclear fractions |
| Band higher than expected | the supplied features do not establish a size-increasing modification | test band identity with MACC1 depletion and an independent antibody |
| Band lower than expected | the supplied features do not establish a smaller mature form | check sample integrity and test band identity with MACC1 depletion |
| Multiple bands | a distinct isoform pattern is not established | compare bands after MACC1 depletion and with an independent antibody |
| Weak or no signal | MACC1 may be unevenly distributed between cytoplasm and nucleus | check both fractions and include a validated positive control |
| Fragments below expected size | sample degradation is possible; no specific MACC1 cleavage is annotated | prepare fresh lysate with protease inhibitors and confirm fragments with a specificity control |
Experiment evidence
Western blot images showing MACC1 band
A04732 · MACC1
A04732 · MACC1
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for MACC1 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for MACC1 in Western blot, you can use adrenal gland tissue, which HPA reports as highly expressed.
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 alongside the samples.
⚠️Feasibility: Whole-tissue lysate is suitable for this intracellular protein, and adipose tissue provides an HPA not-detected comparator.
HPA tissue expression evidence for MACC1
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 → |
| Appendix | endocrine cells | High | Protein (IHC) | HPA → |
| Bronchus | respiratory epithelial cells | High | Protein (IHC) | HPA → |
| Colon | endocrine cells | High | Protein (IHC) | HPA → |
| Duodenum | endocrine 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 → |
| Bone marrow | hematopoietic cells | Low | Protein (IHC) | HPA → |
| Caudate | glial cells | Low | Protein (IHC) | HPA → |
| Cerebellum | cells in granular layer | Low | Protein (IHC) | HPA → |
| Epididymis | glandular cells | Low | Protein (IHC) | HPA → |
Section 3
Advanced MACC1 Western Blot Tips
Deeper troubleshooting and optimisation questions for MACC1, answered from its protein features.
What band size should I expect for MACC1?
Band shift · The predicted mass is 96.6 kDa. No observed Western blot band size is supplied, so use 96.6 kDa as a reference rather than an exact apparent size.
Could MACC1 isoforms produce multiple bands?
Isoforms · The supplied record lists one isoform and no alternative sequence. It provides no basis for assigning additional bands to annotated MACC1 isoforms.
Could phosphorylation explain a shifted MACC1 band?
PTM · UniProt lists phosphoserine at position 19, using its sequence numbering. This establishes a modification site but does not show that phosphorylation causes a visible shift or explain any difference from the predicted 96.6 kDa mass.
Does this guide establish induction of MACC1?
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 MACC1?
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 A04732 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 I quantify MACC1 across samples?
Quantitation · Quantify the band assigned to MACC1 consistently across samples, using the same fraction and normalization approach. Because MACC1 occurs in cytoplasm and nucleus, a change in one fraction need not represent a change in total cellular MACC1.
Should I examine cytoplasmic and nuclear MACC1 separately?
Interpretation · MACC1 is reported in both cytoplasm and nucleus and is mainly cytoplasmic in non-metastasizing tumors. If localization matters, analyze matched cytoplasmic and nuclear fractions and compare each with its corresponding fraction controls.
Compare them with the 96.6 kDa predicted mass and assess band identity experimentally. The supplied features list phosphoserine 19 and one isoform, but neither establishes the cause of an additional band or a mass difference.
Compare them with the 96.6 kDa predicted mass and assess band identity experimentally. The supplied features list phosphoserine 19 and one isoform, but neither establishes the cause of an additional band or a mass difference.
Boster reagents
MACC1 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-MACC1 Antibody
Cat # A04732
The catalog reports A04732 anti-MACC1 antibody with stated Human, Mouse, and Rat reactivity. Its supplied Western blot image shows mouse liver tissue lysate tested at 1 and 2 μg/mL. No human or rat blot is provided.
Which to pick: A04732 is the only listed option. It has a Western blot image using mouse liver lysate; for human or rat samples, the supplied evidence provides stated reactivity but no blot example.
Source: BosterBio MACC1 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q6ZN28.
- Human Protein Atlas. MACC1 tissue expression.
- PMC8805864 — target-verified WB comparison
- PMC12288408 — target-verified WB comparison
- PMC9303487 — target-verified WB comparison