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- Table of Contents
SEC22B / Vesicle-trafficking protein SEC22b · Western blot design guide
Design a Western Blot for SEC22B
Source-linked SEC22B Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-SEC22B 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
SEC22B 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 | ~24.7 kDa | |
| Observed band | ~23 kDa | |
| Gel | 12% (catalog A06284-2) | |
| 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 | 1 isoform(s) |
Section 1
Source-Linked SEC22B Western Blot Protocol Options
The A06284-2 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human U87, human Hela, human MCF-7, human SH-SY5Y, rat brain, rat liver, mouse brain, mouse liver (catalog A06284-2) |
| Gel % | 12% (catalog A06284-2) |
| Load | 30 ug; reducing conditions (catalog A06284-2) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog A06284-2) |
| Membrane | nitrocellulose membrane (catalog A06284-2) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog A06284-2) |
| Primary antibody | A06284-2 · 0.5 μg/mL (catalog A06284-2) |
| Primary incubation | overnight at 4°C (catalog A06284-2) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog A06284-2) |
| Secondary incubation | 1.5 hour at RT (catalog A06284-2) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog A06284-2) |
| Detection | ECL (catalog A06284-2) |
Section 2
What Is the Expected SEC22B Western Blot Band Size?
SEC22B is predicted at 24.7 kDa and observed at ~23 kDa; the cause of the difference is not established.
| Band at ~23 kDa | Empirical SEC22B band in reducing whole-cell lysates; confirm identity with controls |
| Band near 24.7 kDa | Near the sequence-predicted mass, but identity still needs confirmation |
| Weak or absent band in a soluble fraction | SEC22B is a membrane protein and may be enriched in the membrane fraction |
| No distinct modification-dependent doublet | Annotated acetylation and phosphorylation sites do not establish resolvable bands |
💡Expected SEC22B appearanceSEC22B is predicted at 24.7 kDa, while antibody QC detects ~23 kDa in reducing whole-cell lysates; the difference is unexplained by the supplied features, so confirm identity with depletion or an independent antibody.
| Predicted molecular mass | The 215-residue sequence predicts 24.7 kDa; the empirical band is ~23 kDa |
| Lys38 acetylation | The annotated site does not establish a resolvable size shift |
| Ser137, Ser164, Ser168, Ser174 and Ser177 phosphorylation | These annotated sites do not establish distinct band sizes |
| Thr140 phosphorylation | The annotated site does not establish a resolvable size shift |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | SEC22B is membrane-associated and may be poorly recovered | Check membrane recovery and solubilization |
| Band higher than expected | Band identity or migration is uncertain; annotated modifications do not establish a shift | Compare with SEC22B-depleted lysate and an independent antibody |
| Band lower than expected | The ~23 kDa band is empirically observed, but its difference from 24.7 kDa is unexplained | Confirm identity with SEC22B depletion or an independent antibody |
| Broad smear instead of sharp band | Possible incomplete solubilization of membrane-associated SEC22B | Check membrane extraction and sample preparation |
| Multiple bands | Only one isoform is listed; additional band identities are unestablished | Compare bands with SEC22B-depleted lysate |
| Weak or no signal | Low recovery of membrane-associated SEC22B | Examine a membrane-enriched fraction and verify transfer |
Experiment evidence
Western blot images showing SEC22B band
A06284-2 · SEC22B
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for SEC22B Western blot
🧪For positive controls for SEC22B in Western blot, you can use no HPA-supported tissue or cell line from the supplied evidence.
Positive control: No high/medium HPA tissue identified
Negative control: Suggested KO / knockdown lysate
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside the samples.
⚠️Feasibility: HPA provides no expression data, so neither a positive sample nor a clean negative tissue can be verified.
HPA tissue expression evidence for SEC22B
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 |
|---|---|---|---|---|
| No high/medium HPA tissues identified in the supplied evidence. | ||||
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| No lower-expression tissue rows available in the supplied evidence. | ||||
Section 3
Advanced SEC22B Western Blot Tips
Deeper troubleshooting and optimisation questions for SEC22B, answered from its protein features.
How should SEC22B 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 SEC22B isoforms explain multiple bands?
Isoforms · The supplied record lists one isoform and no alternative sequence. It therefore provides no isoform-based explanation for multiple bands.
Which SEC22B modifications should I consider when interpreting bands?
PTM · Using the supplied UniProt coordinates, SEC22B has N6-acetyllysine at 38; phosphoserine at 137, 164, 168, 174 and 177; and phosphothreonine at 140. These sites are candidates to investigate, but their presence alone does not establish a visible band shift.
Does this guide establish induction of SEC22B?
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 SEC22B Western blot?
Transfer · SEC22B is a 215-residue, single-pass membrane protein with a predicted mass of 24.7 kDa. Choose transfer conditions that retain a band near the observed ~23 kDa, then check the membrane and residual gel to assess transfer. The supplied features do not specify one transfer method.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A06284-2 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 the SEC22B band?
Quantitation · Quantify a consistently identified band near the observed ~23 kDa across samples. SEC22B is reported at ER, ER–Golgi intermediate compartment and cis-Golgi membranes, so keep sample preparation consistent when comparing its signal.
Why does SEC22B appear near 23 kDa instead of 24.7 kDa?
Interpretation · The supplied apparent band is ~23 kDa, while the predicted mass is 24.7 kDa. The listed features do not establish the cause of that difference. Compare samples against a molecular weight marker and confirm band identity before attributing the difference to a modification.
The record describes SEC22B as a component of SNARE complexes and lists acetylation and phosphorylation sites. It does not show that complexes or modifications produce extra Western blot bands. Treat additional bands as unassigned until their identity is checked.
The record describes SEC22B as a component of SNARE complexes and lists acetylation and phosphorylation sites. It does not show that complexes or modifications produce extra Western blot bands. Treat additional bands as unassigned until their identity is checked.
Boster reagents
SEC22B 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-SEC22B Antibody Picoband®
Cat # A06284-2
The catalog reports one anti-SEC22B antibody, A06284-2, with reported human, mouse, and rat reactivity. Its Western blot image shows a band near 23 kDa versus an expected 25 kDa in the specified cell and tissue lysates; evidence is limited to the supplied image.
Which to pick: A06284-2 is the only listed choice. Its Western blot image includes human U87, Hela, MCF-7, and SH-SY5Y cells plus rat and mouse brain and liver lysates, using 30 µg per lane and 0.5 µg/mL antibody.
Source: BosterBio SEC22B gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry O75396.
- Human Protein Atlas. SEC22B tissue expression.
- PMC8324795 — target-verified WB comparison
- PMC6685974 — target-verified WB comparison
- PMC10550925 — target-verified WB comparison