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MAP1LC3B / Microtubule-associated protein 1 light chain 3 beta · Western blot design guide
Design a Western Blot for MAP1LC3B
Real validated MAP1LC3B Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-MAP1LC3B 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
MAP1LC3B 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 | ~14.7 kDa | |
| Gel | 15% (standard starting point) | |
| Positive control | Caudate (IHC candidate; verify WB) +4 more | |
| Negative control | Appendix (IHC candidate; verify WB) |
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 | 1 isoform(s) |
Section 1
Real Curated MAP1LC3B Western Blot Protocols
The A01524 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human brain tissue lysate (catalog A01524) |
| Gel % | 15% (standard starting point) |
| Load | 20–30 µg total protein per lane; optimize for abundance (standard starting point) |
| Transfer | Short semi-dry transfer; verify retention (standard starting point) |
| Membrane | 0.2 µm PVDF (standard starting point) |
| Blocking | 5% milk or 5% BSA in TBST (standard starting point) |
| Primary antibody | A01524 · 1 μg/ml (catalog A01524) |
| 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 MAP1LC3B Western Blot Band Size?
MAP1LC3B is predicted at 14.7 kDa; propeptide cleavage and lipid anchoring are annotated, but their effects on observed migration are not established.
| Band near 14.7 kDa | Compatible with the predicted full-length protein; confirm identity with controls |
| Band slightly below 14.7 kDa | Could reflect removal of the 121–125 propeptide |
| Close doublet near 14.7 kDa | Could include precursor and processed protein; band identities need confirmation |
| Band enriched in a membrane fraction | Compatible with the annotated lipid-anchored, autophagosome-associated protein |
💡Expected MAP1LC3B appearanceThe full-length protein is predicted at 14.7 kDa; cleavage of residues 121–125 or lipid anchoring may affect its band, but migration has not been measured here, so confirm identity with controls.
| Predicted full-length mass | Provides a 14.7 kDa reference, not a measured band position |
| 125-residue precursor | Defines the full-length form used for the predicted mass |
| Propeptide at residues 121–125 | Its removal reduces the protein's mass; the apparent shift is unknown |
| Lipid anchor | May affect migration; no band position or shift is supplied |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Membrane-associated protein may be poorly recovered | Check membrane extraction and a positive control |
| Band lower than expected | Processing of the 121–125 propeptide is possible | Compare with a validated MAP1LC3B control |
| Multiple bands | Precursor and processed or lipid-anchored forms are possible, but their separation is unestablished | Use a validated positive control and compare sample fractions |
| Weak or no signal | Membrane-associated protein may be underrepresented in the preparation | Check extraction, loading, and antibody performance with a positive control |
| Fragments below expected size | Sample degradation or cleavage beyond the annotated propeptide may have occurred | Repeat with protease inhibition and compare with a fresh positive control |
Experiment evidence
Western blot images showing MAP1LC3B band
A01524 · MAP1LC3B
A01524T50 · MAP1LC3B
A01524T50 · MAP1LC3B
A01524-2 · MAP1LC3B
M01524 · MAP1LC3B
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for MAP1LC3B Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for MAP1LC3B in Western blot, you can use caudate tissue, which HPA rates as high expression.
Positive control: Caudate (IHC candidate; verify WB)
Negative control: Appendix (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain such as Ponceau alongside the samples.
⚠️Feasibility: HPA reports appendix as not detected, providing a feasible tissue negative control.
HPA tissue expression evidence for MAP1LC3B
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 |
|---|---|---|---|---|
| Caudate | neuronal cells | High | Protein (IHC) | HPA → |
| Thyroid gland | glandular cells | High | Protein (IHC) | HPA → |
| Adrenal gland | glandular cells | Medium | Protein (IHC) | HPA → |
| Breast | glandular cells | Medium | Protein (IHC) | HPA → |
| Bronchus | respiratory epithelial cells | Medium | Protein (IHC) | HPA → |
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Appendix | glandular cells | Not detected | Protein (IHC) | HPA → |
| Colon | endothelial cells | Not detected | Protein (IHC) | HPA → |
| Esophagus | squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
| Fallopian tube | glandular cells | Not detected | Protein (IHC) | HPA → |
| Heart muscle | cardiomyocytes | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced MAP1LC3B Western Blot Tips
Deeper troubleshooting and optimisation questions for MAP1LC3B, answered from its protein features.
How should MAP1LC3B 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.
Should I expect multiple MAP1LC3B isoforms?
Isoforms · The supplied UniProt record lists one isoform and no alternative sequence. Multiple bands should therefore not be assigned to MAP1LC3B splice isoforms on this evidence alone.
Do annotated modifications prove a band shift?
PTM · The linked UniProt record describes protein features. A modification annotation alone does not demonstrate a visible shift; retain any condition or experimental qualifier attached to it.
How should autophagy induction affect MAP1LC3B analysis?
Induction · MAP1LC3B is associated with autophagy and the autophagosome membrane, and UniProt lists a lipid anchor. Compare matched samples with consistent collection times and consider both soluble and membrane-associated forms. A change in one band alone does not establish increased autophagic flux.
What transfer method to use for MAP1LC3B Western blot?
Transfer · MAP1LC3B has a predicted mass of 14.7 kDa, so check transfer conditions for retention of small proteins. Confirm transfer with a low-molecular-weight marker and avoid conditions that let the target pass through the membrane.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A01524 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 MAP1LC3B bands be quantified?
Quantitation · Quantify each reproducible band separately using the same exposure and a defined loading normalization method. The record identifies MAP1LC3B with autophagy, an autophagosome membrane location, and a lipid anchor; a single band intensity cannot by itself measure autophagic flux.
Why might MAP1LC3B migrate differently from its predicted mass?
Interpretation · The predicted mass is 14.7 kDa for the 125-residue sequence. UniProt lists a propeptide at residues 121..125 and a lipid anchor, either of which could affect the form being measured. These features alone do not establish a visible shift or explain a particular band position.
UniProt marks residues 121..125 as a propeptide. An antibody recognizing this segment may detect the precursor differently from a processed form. Check the antibody epitope before assigning bands; the listed coordinates use UniProt numbering.
Consider the propeptide at UniProt residues 121..125, lipid anchoring, and association with MAP1A or MAP1B heavy chains when evaluating band identity. None of these features proves that a specific extra band is MAP1LC3B. Check antibody specificity and compare band behavior across samples before assigning it.
UniProt marks residues 121..125 as a propeptide. An antibody recognizing this segment may detect the precursor differently from a processed form. Check the antibody epitope before assigning bands; the listed coordinates use UniProt numbering.
Consider the propeptide at UniProt residues 121..125, lipid anchoring, and association with MAP1A or MAP1B heavy chains when evaluating band identity. None of these features proves that a specific extra band is MAP1LC3B. Check antibody specificity and compare band behavior across samples before assigning it.
Boster reagents
MAP1LC3B 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-MAP1LC3B Antibody
Cat # A01524
Real WB data
Anti-LC3 (T50) MAP1LC3B Antibody
Cat # A01524T50
Real WB data
Anti-LC3B/MAP1LC3B Antibody
Cat # A01524-2
Real WB data
Anti-LC3B MAP1LC3B Rabbit Monoclonal Antibody
Cat # M01524
Four the supplier anti-MAP1LC3B antibodies list human, mouse, and rat reactivity and have WB images. Their captions document specific lysates and conditions; these examples do not establish performance in every sample type or independent validation.
Which to pick: All four have WB images. Choose A01524 for a human brain example, A01524T50 for cell lysate examples, or A01524-2 or M01524 for documented human cell and rat and mouse brain lanes. M01524 is monoclonal.
Source: BosterBio MAP1LC3B gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q9GZQ8.
- Human Protein Atlas. MAP1LC3B tissue expression.
- PMC4264866 — target-verified WB comparison
- PMC9225222 — target-verified WB comparison
- PMC6430800 — target-verified WB comparison