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- Table of Contents
SEC13 / Protein SEC13 homolog · Western blot design guide
Design a Western Blot for SEC13
Real validated SEC13 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-SEC13 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
SEC13 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 | ~35.5 kDa | |
| Observed band | ~36 kDa | |
| Gel | 5–20% (catalog A04346-1) | |
| Positive control | Breast (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 | 4 isoform(s) |
Section 1
Real Curated SEC13 Western Blot Protocols
The A04346-1 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human MCF-7, human MDA-MB-453, human HepG2 (catalog A04346-1) |
| Gel % | 5–20% (catalog A04346-1) |
| Load | 30 ug; reducing conditions (catalog A04346-1) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog A04346-1) |
| Membrane | nitrocellulose membrane (catalog A04346-1) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog A04346-1) |
| Primary antibody | A04346-1 · 0.5 μg/mL (catalog A04346-1) |
| Primary incubation | overnight at 4°C (catalog A04346-1) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog A04346-1) |
| Secondary incubation | 1.5 hour at RT (catalog A04346-1) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog A04346-1) |
| Detection | ECL (catalog A04346-1) |
Section 2
What Is the Expected SEC13 Western Blot Band Size?
SEC13 is predicted at 35.5 kDa and observed at ~36 kDa; the supplied evidence does not establish a cause for the small difference.
| Band at ~36 kDa | Empirical SEC13 band, close to the 35.5 kDa predicted mass |
| Additional band above ~36 kDa | Could represent an SEC13 isoform; its migration is unestablished |
| Additional band below ~36 kDa | Could represent an SEC13 isoform; its migration is unestablished |
| Several bands of different sizes | Could reflect isoforms 1, 2, 3, and 4; distinct bands are unconfirmed |
💡Expected SEC13 appearanceSEC13 has a predicted mass of 35.5 kDa and an empirical band at ~36 kDa; confirm band identity with appropriate controls, since the supplied features do not establish distinct isoform migration.
| Predicted SEC13 mass | 35.5 kDa, close to the empirical ~36 kDa band |
| Splice isoform 1 | Its individual apparent size is not supplied |
| Splice isoform 2 | May differ in size from other isoforms; its migration is not supplied |
| Splice isoform 3 | May differ in size from other isoforms; its migration is not supplied |
| Splice isoform 4 | May differ in size from other isoforms; its migration is not supplied |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | SEC13 may be poorly recovered from its membrane-associated or nuclear pore locations | Check extraction and a positive-control lysate |
| Band higher than expected | An isoform is possible, but its migration is unknown | Compare with the ~36 kDa reference band and verify identity with a second antibody |
| Band lower than expected | An isoform is possible, but its migration is unknown | Check band identity with a second antibody or SEC13 depletion |
| Multiple bands | Four SEC13 isoforms are annotated, but distinct migration is unconfirmed | Compare isoform expression and verify bands with SEC13 depletion |
| Weak or no signal | Recovery from SEC13's membrane-associated or nuclear pore locations may be limited | Check lysate preparation, loading, and a positive control |
Experiment evidence
Western blot images showing SEC13 band
A04346-1 · SEC13
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for SEC13 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for SEC13 in Western blot, you can use breast tissue, which HPA scores High.
Positive control: Breast (IHC candidate; verify WB)
Negative control: Adipose tissue (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside SEC13.
⚠️Feasibility: HPA reports SEC13 as not detected in adipose tissue, but confirm that it is negative by Western blot.
HPA tissue expression evidence for SEC13
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 |
|---|---|---|---|---|
| Breast | glandular cells | High | Protein (IHC) | HPA → |
| Cervix | squamous epithelial cells | High | Protein (IHC) | HPA → |
| Esophagus | squamous epithelial cells | High | Protein (IHC) | HPA → |
| Prostate | glandular cells | High | Protein (IHC) | HPA → |
| Tonsil | squamous epithelial 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 → |
| Cerebellum | cells in granular layer | Not detected | Protein (IHC) | HPA → |
| Hippocampus | glial cells | Not detected | Protein (IHC) | HPA → |
| Kidney | cells in glomeruli | Not detected | Protein (IHC) | HPA → |
| Liver | cholangiocytes | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced SEC13 Western Blot Tips
Deeper troubleshooting and optimisation questions for SEC13, answered from its protein features.
How should SEC13 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 SEC13 isoforms produce different bands?
Isoforms · Four isoforms are listed. Relative to the canonical sequence, isoform 2 lacks residues 1–14; isoform 3 replaces the initial methionine with a longer sequence; and isoform 4 replaces it with MGKM. These differences could affect apparent size, but the features do not establish which isoforms appear on a blot.
Which SEC13 modifications are listed?
PTM · UniProt lists N-acetylvaline at position 2 and phosphoserine at positions 184 and 309. These are UniProt coordinates for the supplied sequence; isoform or antibody numbering may differ. The features do not identify a modifying enzyme or an inducing condition.
No. Phosphoserine at UniProt positions 184 and 309 documents modification sites, but does not show that phosphorylation causes a resolvable mobility shift. Do not assign a shifted band to phosphorylation from position data alone.
No. Phosphoserine at UniProt positions 184 and 309 documents modification sites, but does not show that phosphorylation causes a resolvable mobility shift. Do not assign a shifted band to phosphorylation from position data alone.
Does this guide establish induction of SEC13?
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 SEC13?
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 A04346-1 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 SEC13 bands be quantified across fractions?
Quantitation · Compare matched fractions and normalize within each fraction. Because SEC13 is associated with vesicle and endoplasmic reticulum membranes and the nuclear pore, a change in one fraction alone does not establish a change in total cellular SEC13.
Why is SEC13 observed near 36 kDa?
Interpretation · The observed band near 36 kDa is close to the predicted 35.5 kDa for canonical SEC13. The supplied features do not establish a meaningful mass shift.
SEC13 is listed on the cytoplasmic side of COPII-coated vesicle and endoplasmic reticulum membranes as a peripheral membrane protein; it is also a component of a nuclear pore subcomplex. Include the relevant cellular fraction when preparing samples, and keep fractionation consistent across comparisons.
Check whether their sizes are compatible with the listed isoform sequence differences, then assess band specificity experimentally. The listed acetylation and phosphorylation sites alone cannot identify an extra band or explain its mobility.
SEC13 is listed on the cytoplasmic side of COPII-coated vesicle and endoplasmic reticulum membranes as a peripheral membrane protein; it is also a component of a nuclear pore subcomplex. Include the relevant cellular fraction when preparing samples, and keep fractionation consistent across comparisons.
Check whether their sizes are compatible with the listed isoform sequence differences, then assess band specificity experimentally. The listed acetylation and phosphorylation sites alone cannot identify an extra band or explain its mobility.
Boster reagents
SEC13 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-SEC13L1/SEC13 Antibody Picoband®
Cat # A04346-1
The catalog reports one anti-SEC13 antibody for Western blotting, A04346-1. Its WB image reports a band near the expected 36 kDa in human cell, rat tissue, and mouse tissue lysates. The supplied evidence is a product validation image; no publication evidence is provided.
Which to pick: A04346-1 is the only listed option. It reports human, mouse, and rat reactivity, with a WB image using human cell lines and rat and mouse brain and small intestine lysates. Match your sample and conditions to those shown.
Source: BosterBio SEC13 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P55735.
- Human Protein Atlas. SEC13 tissue expression.
- PMC8887674 — target-verified WB comparison
- PMC8223750 — target-verified WB comparison
- PMC2922642 — target-verified WB comparison