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- Table of Contents
MAP1B / Microtubule-associated protein 1B · Western blot design guide
Design a Western Blot for MAP1B
Real validated MAP1B Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-MAP1B 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
MAP1B 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 | ~270.6 kDa | |
| Observed band | ~310 kDa | |
| Gel | 8% (catalog A01760-1) | |
| Positive control | Cerebellum (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 Hallmark membership | |
| Isoform | 1 isoform(s) |
Section 1
Real Curated MAP1B Western Blot Protocols
The A01760-1 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human A549, human U251, human SH-SY5Y (catalog A01760-1) |
| Gel % | 8% (catalog A01760-1) |
| Load | 30 ug; reducing conditions (catalog A01760-1) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog A01760-1) |
| Membrane | nitrocellulose membrane (catalog A01760-1) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog A01760-1) |
| Primary antibody | A01760-1 · 0.5 μg/mL (catalog A01760-1) |
| Primary incubation | overnight at 4°C (catalog A01760-1) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog A01760-1) |
| Secondary incubation | 1.5 hour at RT (catalog A01760-1) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog A01760-1) |
| Detection | ECL (catalog A01760-1) |
Section 2
What Is the Expected MAP1B Western Blot Band Size?
MAP1B is predicted at 270.6 kDa, while antibody QC reports ~310 kDa; the cause of this difference is not established.
| Band near 310 kDa | Empirical MAP1B band reported in whole-cell and brain lysates; confirm identity with controls |
| Band near 271 kDa | Near the calculated 270.6 kDa mass of full-length MAP1B; identity requires confirmation |
| Lower band | Could represent proteolytic processing, including the MAP1B-derived LC1 light chain; fragment mass is unspecified |
| Doublet near the main band | Could reflect different phosphorylation states; distinct migration is not established |
💡Expected MAP1B appearanceFull-length MAP1B has a predicted mass of 270.6 kDa, while antibody QC reports a band near 310 kDa; the cause of that difference is unestablished, so confirm band identity with appropriate controls.
| Full-length predicted mass | Calculated mass is 270.6 kDa; the reported empirical band is near 310 kDa |
| Phosphoserines at residues 336, 339, and 343 | Phosphorylation may affect migration, but no size shift is established |
| Phosphothreonine at residue 527 | Phosphorylation may affect migration, but no size shift is established |
| Proteolytic production of LC1 | Produces a MAP1B-derived light chain; its apparent mass is not supplied |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | MAP1B is associated with the cytoskeleton, and recovery may vary with extraction | Check an appropriate positive lysate and inspect the insoluble fraction |
| Band higher than expected | The reported ~310 kDa band exceeds the 270.6 kDa calculated mass; its migration difference is unexplained | Compare with a validated positive lysate and verify band identity |
| Band lower than expected | MAP1B can undergo proteolytic processing to yield LC1 | Check antibody epitope coverage and compare with a full-length MAP1B control |
| Multiple bands | Proteolytic products or different phosphorylation states are possible, but band identities are unestablished | Compare antibody epitopes and test phosphatase treatment with suitable controls |
| Weak or no signal | Transfer or extraction of large cytoskeletal MAP1B may be inefficient | Check transfer efficiency and include a positive lysate |
| Fragments below expected size | LC1 production or sample proteolysis may yield smaller immunoreactive species | Check epitope location and repeat with protease inhibitors |
Experiment evidence
Western blot images showing MAP1B band
A01760-1 · MAP1B
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for MAP1B Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for MAP1B in Western blot, you can use cerebellum lysate, which HPA reports as highly positive.
Positive control: Cerebellum (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: Tissue lysates are suitable for cytoskeletal MAP1B, and HPA reports adipose tissue as not detected.
HPA tissue expression evidence for MAP1B
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 | GLUC cells - cytoplasm/membrane | High | Protein (IHC) | HPA → |
| Cerebral cortex | neuronal cells | High | Protein (IHC) | HPA → |
| Colon | peripheral nerve/ganglion | High | Protein (IHC) | HPA → |
| Placenta | decidual cells | High | Protein (IHC) | HPA → |
| Rectum | peripheral nerve/ganglion | 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 → |
| Appendix | endocrine cells | Not detected | Protein (IHC) | HPA → |
| Bone marrow | hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Breast | adipocytes | Not detected | Protein (IHC) | HPA → |
| Caudate | glial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced MAP1B Western Blot Tips
Deeper troubleshooting and optimisation questions for MAP1B, answered from its protein features.
How should MAP1B 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.
Are alternative isoforms expected from these features?
Isoforms · The supplied record lists one isoform and no alternative sequence. It therefore provides no basis for assigning an extra band to a named alternative isoform. Consider the documented LC1 cleavage product when assessing smaller bands.
Which modifications matter when comparing MAP1B bands?
PTM · UniProt lists numerous phosphorylation sites, including Ser336, Ser339, Ser343, Tyr1410 and Tyr1796, plus N-acetylalanine at position 2 and S-nitrosocysteine at position 2464. These are UniProt sequence coordinates; antibody or paper numbering may differ. Compare modification-specific signals only when the antibody targets a listed site, and do not infer a visible shift from site annotation alone.
Does this guide establish induction of MAP1B?
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 MAP1B Western blot?
Transfer · MAP1B is a large protein with a predicted mass of 270.6 kDa and a reported apparent band near 310 kDa. Choose a transfer setup validated for proteins in that size range, and check transfer at the high molecular weight region before interpreting a weak or absent band.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A01760-1 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 MAP1B band intensity be quantified?
Quantitation · Quantify the same defined band region across samples and report whether the measurement represents the ~310 kDa apparent band or another verified MAP1B species. Because MAP1B has many annotated phosphorylation sites and a documented LC1 cleavage product, do not combine distinct bands without establishing what each represents.
Why might MAP1B appear near 310 kDa?
Interpretation · The supplied apparent band is ~310 kDa, while the predicted mass is 270.6 kDa. MAP1B has many annotated modifications, but their presence alone does not establish the cause of this difference. Confirm band identity before assigning a mechanism.
MAP1B is reported to produce light chain LC1 by cleavage, which can associate with MAP1B heavy chains. A smaller band could therefore be investigated as a cleavage product; its size alone does not establish its identity. Check whether the antibody recognizes the region retained in LC1.
MAP1B is reported to produce light chain LC1 by cleavage, which can associate with MAP1B heavy chains. A smaller band could therefore be investigated as a cleavage product; its size alone does not establish its identity. Check whether the antibody recognizes the region retained in LC1.
Boster reagents
MAP1B 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-MAP1B Antibody Picoband®
Cat # A01760-1
The catalog reports one anti-MAP1B antibody for Western blotting, A01760-1, with stated human, mouse, and rat reactivity. Its WB image includes human cell, rat brain and cell, and mouse cell lysates; the reported band is approximately 310 kDa versus 271 kDa expected.
Which to pick: A01760-1 is the only listed option and has a WB image across the named human, rat, and mouse samples. Check whether its reported approximately 310 kDa band fits your assay before choosing it.
Source: BosterBio MAP1B gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P46821.
- Human Protein Atlas. MAP1B tissue expression.
- PMC10661827 — target-verified WB comparison
- PMC4211189 — target-verified WB comparison
- PMC4948024 — target-verified WB comparison