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SEC62 / Translocation protein SEC62 · Western blot design guide
Design a Western Blot for SEC62
Source-linked SEC62 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-SEC62 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
SEC62 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 | ~45.9 kDa | |
| Gel | 12–15% (standard starting point) | |
| Positive control | Bone marrow (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 SEC62 Western Blot Protocol Options
The A05906 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | rat brain tissue lysate (catalog A05906) |
| Gel % | 12–15% (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 | A05906 · (A) 0.5 and (B) 1 μg/mL (catalog A05906) |
| 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 SEC62 Western Blot Band Size?
SEC62 is predicted at 45.9 kDa; membrane association and phosphorylation may affect detection or migration, but no empirical band position or migration effect is established.
| Band near 45.9 kDa | Consistent with the UniProt predicted mass; confirm SEC62 identity with controls |
| Weak or absent band in whole-cell lysate | SEC62 may be poorly recovered because it is an ER membrane protein |
| Band above 45.9 kDa | SEC62 identity needs confirmation; the listed phosphorylation sites do not establish a visible shift |
| Doublet near 45.9 kDa | Distinct phosphorylation states are possible, but their migration is not established |
💡Expected SEC62 appearanceSEC62 has a predicted mass of 45.9 kDa and no supplied empirical band size; use antibody specificity and sample controls to identify a candidate band.
| UniProt predicted mass | Provides a 45.9 kDa reference, not a measured band position |
| Phosphothreonine at residue 158 | Could alter migration, but no visible shift is established |
| Phosphoserine at residue 335 | Could alter migration, but no visible shift is established |
| Phosphoserine at residue 341 | Could alter migration, but no visible shift is established |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | Membrane-associated SEC62 may be poorly extracted | Check membrane protein recovery and use a positive lysate control |
| Band higher than expected | The supplied features do not establish the cause | Confirm band identity with an independent antibody or SEC62 depletion |
| Band lower than expected | The supplied features do not establish the cause | Check sample integrity and confirm band identity |
| Multiple bands | Phosphorylation is documented, but distinct migrating forms are unproven | Compare with a phosphatase-treated sample and verify band identity |
| Weak or no signal | Recovery of ER membrane SEC62 may be low | Check extraction, loading, and a positive control |
Experiment evidence
Western blot images showing SEC62 band
A05906 · SEC62
M05906 · SEC62
M05906 · SEC62
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for SEC62 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for SEC62 in Western blot, you can use bone marrow tissue, which HPA rates High.
Positive control: Bone marrow (IHC candidate; verify WB)
Negative control: Adipose tissue (IHC candidate; verify WB)
Loading controls: Run GAPDH, β-actin, and a total-protein stain such as Ponceau alongside the samples.
⚠️Feasibility: SEC62 is an ER membrane protein, so effective membrane solubilization may be needed for a clear signal.
HPA tissue expression evidence for SEC62
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 |
|---|---|---|---|---|
| Bone marrow | hematopoietic cells | High | Protein (IHC) | HPA → |
| Cerebellum | Purkinje cells | High | Protein (IHC) | HPA → |
| Cervix | glandular cells | High | Protein (IHC) | HPA → |
| Epididymis | glandular cells | High | Protein (IHC) | HPA → |
| Salivary gland | glandular 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 → |
| Skeletal muscle | myocytes | Not detected | Protein (IHC) | HPA → |
| Urinary bladder | urothelial cells | Not detected | Protein (IHC) | HPA → |
| Heart muscle | cardiomyocytes | Low | Protein (IHC) | HPA → |
| Hippocampus | neuronal cells | Low | Protein (IHC) | HPA → |
Section 3
Advanced SEC62 Western Blot Tips
Deeper troubleshooting and optimisation questions for SEC62, answered from its protein features.
How should SEC62 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 SEC62 isoforms explain multiple bands?
Isoforms · The supplied record lists one isoform and no alternative sequence. It does not support assigning additional bands to SEC62 isoforms; verify their identity before interpreting them.
Which SEC62 phosphorylation sites should I consider?
PTM · UniProt lists Thr158, Ser335, Ser341, Ser353, Ser356 and Thr375 as phosphorylated residues. These are UniProt coordinates; antibody or paper numbering may differ. Their presence does not establish that phosphorylation causes a visible band shift.
Does this guide establish induction of SEC62?
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 SEC62 Western blot?
Transfer · SEC62 is a 45.9 kDa, multi-pass ER membrane protein. Use a transfer procedure validated for a membrane protein of this size and check transfer efficiency. The supplied features do not specify wet versus semi-dry transfer or particular settings.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A05906 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 SEC62 bands?
Quantitation · Quantify a verified SEC62 band under comparable sample preparation and detection conditions. Because SEC62 is an ER membrane protein, keep membrane protein recovery consistent across samples. The supplied features provide no observed band position, so confirm band identity before measuring it.
Should SEC62 run at its predicted 45.9 kDa?
Interpretation · 45.9 kDa is the predicted mass; no observed band position is supplied. Use it as a starting point for band identification, but do not assume an apparent mass difference is explained by the listed features.
Confirm band identity before assigning a cause. The record lists one isoform, no glycosylation sites and six phosphorylation sites. Comparing samples with and without phosphatase treatment could test whether phosphorylation contributes to a pattern, but the listed sites alone do not predict a visible shift.
Confirm band identity before assigning a cause. The record lists one isoform, no glycosylation sites and six phosphorylation sites. Comparing samples with and without phosphatase treatment could test whether phosphorylation contributes to a pattern, but the listed sites alone do not predict a visible shift.
Boster reagents
SEC62 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-Translocation protein SEC62 SEC62 Antibody
Cat # A05906
Real WB data
Anti-SEC62 Rabbit Monoclonal Antibody
Cat # M05906
The catalog reports two anti-SEC62 antibodies with Western blot images. A05906 was shown with rat brain lysate at 0.5 and 1 μg/mL; M05906 was shown with five human cell lines and two mouse tissues, detecting a band near 50 kDa. These examples do not establish performance in other samples.
Which to pick: Both list Human, Mouse, and Rat reactivity. Choose A05906 for the documented rat brain example; choose M05906 for the documented human cell line or mouse lung and spleen examples. Both have WB images, but the supplied M05906 caption provides more protocol detail.
Source: BosterBio SEC62 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q99442.
- Human Protein Atlas. SEC62 tissue expression.
- PMC3878975 — target-verified WB comparison
- PMC5354881 — target-verified WB comparison
- PMC4995743 — target-verified WB comparison