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SERPINA7 · Western blot design guide
Design a Western Blot for SERPINA7
Real validated SERPINA7 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-SERPINA7 WB antibodies. Everything you need to plan the experiment before you commit precious samples.
Last reviewed: May 2026 · Scientific review: Boster Bio technical team
SERPINA7 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 | ~46.3 kDa | |
| Observed band | ~50 kDa | |
| Gel | 10–12% | |
| Positive control | Adipose tissue+4 more · see all | |
| Negative control | Adipose tissue+4 more · see all |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Glycosylated + Cleaved | |
| Caveat | N-glycosylation increases mass | |
| Regulation | IFN-γ-induced | |
| Isoform | 1 isoform(s) |
Section 1
Real Curated SERPINA7 Western Blot Protocols
Literature-validated Western blot parameters for SERPINA7 — gel percentage, transfer, blocking, antibody incubation and detection, extracted from published methods.
| Sample / lysate | human HepG2 . After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-Thyroxine Binding Globulin/SERPINA7 antigen affinity purified polyclonal antibody (Catalog # A02766-1) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for Thyroxine Binding Globulin/SERPINA7 at approximately 50 kDa. The expected band size for Thyroxine Binding Globulin/SERPINA7 is at 46 kDa |
| Gel % | 10–12% |
| Load | 30 ug |
| Transfer | nitrocellulose membrane, 150 mA, 50–90 min |
| Membrane | nitrocellulose |
| Blocking | 5% non-fat milk / TBS, 1.5 h RT |
| Primary antibody | 0.5 µg/mL |
| Primary incubation | overnight at 4 °C |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 |
| Wash | TBS-0.1% Tween, 3 × 5 min |
| Detection | ECL |
| Exposure / imaging | Tanon 5200 |
| Observed band | 50 kDa |
Section 2
What Is the Expected SERPINA7 Western Blot Band Size?
SERPINA7/TBG has a 46.3 kDa protein backbone but runs at ~50 kDa on blots because signal-peptide cleavage is outweighed by glycosylation at 5 N-linked sites.
| predominant band at ~50 kDa | the mature, signal-peptide-cleaved, fully N-glycosylated TBG monomer — the expected native band |
| diffuse or smeared band around 48-52 kDa rather than one sharp line | heterogeneous glycan occupancy/branching across the 5 N-linked sites (Asn36, Asn99, Asn116, Asn165, Asn253) broadens the apparent mass |
| sharper band nearer 44-46 kDa | reduced glycan mass, e.g. after deglycosylation treatment or in a non-glycosylated recombinant standard, closer to the unmodified backbone |
| little to no band in whole-cell or tissue lysate | TBG is a secreted plasma protein, so intracellular lysates capture only the pre-secretion pool; serum/plasma is the better sample |
| single clean band without a doublet or extra bands | only one isoform is annotated for SERPINA7, so multiple closely spaced bands are not expected from isoform variation |
💡Expected SERPINA7 appearanceExpect a single band at approximately 50 kDa, reflecting the 46.3 kDa signal-peptide-cleaved TBG backbone carrying added mass from its 5 N-linked glycosylation sites, rather than a sharp 46.3 kDa band.
| Predicted mass (UniProt) | gives the unmodified 46.3 kDa polypeptide backbone for the 415-aa precursor as the starting reference point |
| Signal peptide (residues 1-20) | cleaved during secretion, so the mature circulating protein starts from a slightly smaller backbone than the full-length 46.3 kDa precursor before glycosylation is added |
| N-glycosylation at Asn36, Asn99, Asn116, Asn165 and Asn253 | five complex N-glycans add substantial mass, shifting the mature band up to the ~50 kDa observed on blots and broadening it into a smear when glycoform occupancy varies |
| Secreted subcellular localization | protein is exported from the cell, so whole-cell lysates give weak signal compared with serum or plasma |
| Non-glycosylated recombinant or bacterial standard | lacks the native glycan load and runs closer to the unmodified ~44-46 kDa backbone mass, lower than native plasma TBG |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | TBG is secreted into plasma rather than retained intracellularly | use serum or plasma instead of whole-cell lysate, or confirm only the small pre-secretion pool is being probed |
| Band higher than expected | full or partial occupancy across the 5 N-linked glycosylation sites adds more mass than the 46.3 kDa backbone alone | treat a duplicate sample with PNGase F and check for a shift down toward ~44-46 kDa to confirm glycosylation |
| Band lower than expected | sample is non-glycosylated recombinant protein or has been enzymatically deglycosylated | run a native plasma or serum sample in parallel as a fully glycosylated comparator |
| Broad smear instead of sharp band | heterogeneous glycan branching and occupancy across the 5 N-glycosylation sites | resolve on a lower-percentage or gradient gel and allow a longer run/transfer time |
| Fragments below expected size | loss of the 20-residue signal peptide during normal maturation, or proteolytic degradation of the sample | add protease inhibitors during sample preparation and compare against a fresh, properly stored plasma sample |
Experiment evidence
Western blot images showing SERPINA7 band
A02766-1 · SERPINA7
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for SERPINA7 Western blot
🧪For positive controls for SERPINA7 in Western blot, you can use human serum or plasma, since SERPINA7 is a secreted protein and none of the sampled HPA tissues (adipose tissue, adrenal gland, appendix, bone marrow) showed detectable expression, making biofluid rather than tissue lysate the most reliable positive source.
Positive control: Serum/plasma
Negative control: Adipose tissue
Loading controls: Run GAPDH and β-actin as loading controls alongside a total-protein stain (stain-free gel, Ponceau S, or REVERT) to confirm equal loading.
⚠️Feasibility: As a secreted plasma protein with no detectable signal in the sampled HPA tissue lysates, SERPINA7/TBG is best detected using serum, plasma, or conditioned medium rather than standard whole-cell tissue lysate for a reliable positive control.
HPA tissue expression evidence for SERPINA7
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.
Positive expression · recommended positive controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | adipocytes | Not detected | Protein (IHC) | HPA → |
| Adrenal gland | glandular cells | Not detected | Protein (IHC) | HPA → |
| Appendix | glandular cells | Not detected | Protein (IHC) | HPA → |
| Bone marrow | hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Breast | adipocytes | Not detected | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | adipocytes | Not detected | Protein (IHC) | HPA → |
| Adrenal gland | glandular cells | Not detected | Protein (IHC) | HPA → |
| Appendix | glandular cells | Not detected | Protein (IHC) | HPA → |
| Bone marrow | hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Breast | adipocytes | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced SERPINA7 Western Blot Tips
Deeper troubleshooting and optimisation questions for SERPINA7, answered from its protein features.
Why does SERPINA7 run higher than its predicted 46.3 kDa mass?
The 46.3 kDa prediction is for the unmodified sequence, but SERPINA7 is secreted after removal of a 20-residue signal peptide and carries 5 N-glycosylation sites. Added glycan mass typically shifts the apparent molecular weight upward, so an observed band around 50 kDa on SDS-PAGE is consistent with the mature glycosylated protein, not a loading or antibody issue.
Does glycosylation state affect SERPINA7 band mobility?
SERPINA7 carries 5 predicted N-glycosylation sites and no other listed modifications. Differences in glycan occupancy or sialylation between plasma samples can shift the apparent band by a few kDa, so treat minor mobility variation between samples as normal glycoform heterogeneity rather than a specificity or antibody problem.
Is SERPINA7 expression hormonally regulated?
SERPINA7 (thyroxine-binding globulin) is a secreted glycoprotein, and its circulating levels and glycosylation are known to shift with estrogen status. When comparing samples across hormonal conditions, expect changes in signal intensity and slight migration differences that reflect glycoform and abundance changes rather than antibody performance issues.
What blocking buffer works best for SERPINA7 detection?
Because SERPINA7 is a glycoprotein, avoid glycoprotein-rich milk-based blocking if using a lectin-based or glycan-sensitive detection step, since milk components can cross-react. A standard BSA-based block is a safer default for this secreted glycoprotein and reduces background without masking glycan-dependent epitopes some antibodies recognize.
What transfer method to use for SERPINA7 Western blot?
At roughly 50 kDa, SERPINA7 transfers efficiently with standard wet or semi-dry transfer protocols. Because it is a glycosylated secreted protein, PVDF membrane is preferred over nitrocellulose for better retention of glycoproteins, and standard transfer times for mid-size proteins are sufficient without needing extended transfer conditions.
How should SERPINA7 signal be normalized across serum samples?
SERPINA7 is a secreted plasma protein, so intracellular loading controls like actin or GAPDH are not appropriate for serum or plasma samples. Use total protein normalization, such as stain-free imaging or Ponceau staining, instead, since housekeeping proteins are absent from serum and cannot serve as a valid reference.
What causes extra bands in a SERPINA7 blot?
SERPINA7 has no disulfide bonds and no known oligomeric assembly, so it should appear as a single monomeric band near 50 kDa under both reducing and non-reducing conditions. Higher molecular weight bands likely indicate nonspecific aggregation, while lower bands can reflect proteolytic degradation of this secreted plasma protein during sample handling.
Boster reagents
Best SERPINA7 Western Blot Antibodies
BosterBio's SERPINA7 antibodies are among the best-performing WB antibodies on the market — well cited, thoroughly validated, and orthogonally cross-validated against negative tissues and complementary methods.
Real WB data
Anti-Thyroxine Binding Globulin/SERPINA7 Antibody Picoband®
Cat # A02766-1
The recommended anti-SERPINA7 (Thyroxine Binding Globulin) antibody is a top-performing, widely cited reagent with thorough Western blot validation, including orthogonal confirmation against negative tissue and complementary detection approaches, giving confidence in its specificity and reproducibility for reliable SERPINA7 detection.
Which to pick: Only one Boster SERPINA7 antibody is catalogued, A02766-1, and it includes an actual Western blot validation image (5-20% SDS-PAGE, 70V), so it is both the recommended and the only available choice for WB work on this target.
Source: BosterBio SERPINA7 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P05543.
- Human Protein Atlas. SERPINA7 tissue expression.