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- Table of Contents
AZGP1 / Zinc-alpha-2-glycoprotein · Western blot design guide
Design a Western Blot for AZGP1
Source-linked AZGP1 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-AZGP1 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
AZGP1 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 | ~34.3 kDa | |
| Gel | 12–15% (standard starting point) | |
| Positive control | Appendix (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 | Glycosylated + Cleaved | |
| Caveat | Glycosylation-state controls | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 1 isoform(s) |
Section 1
Source-Linked AZGP1 Western Blot Protocol Options
The M02718-1 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | HepG2 cell lysate (catalog M02718-1) |
| 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 | M02718-1; use the WB datasheet starting dilution (standard starting point) |
| 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 AZGP1 Western Blot Band Size?
AZGP1 has a predicted 34.3 kDa precursor; signal-peptide cleavage and N-linked glycosylation may affect migration, but no empirical band size is supplied.
| Band near 34.3 kDa | consistent with the predicted precursor mass; confirm identity with a control |
| Band above 34.3 kDa | N-linked glycans may increase apparent size |
| Band below 34.3 kDa | signal-peptide cleavage may contribute to a smaller mature form |
| Little or no band in whole-cell lysate | AZGP1 is secreted |
| Broad band or smear | variation in N-linked glycans may contribute |
💡Expected AZGP1 appearanceUniProt predicts a 34.3 kDa precursor, but signal-peptide cleavage and N-linked glycosylation may alter native migration; no empirical band size is supplied, so confirm band identity with controls.
| Predicted precursor mass | 34.3 kDa is the sequence-based reference, not a measured band |
| Signal peptide at residues 1–20 | cleavage can reduce the mature polypeptide mass |
| N-linked glycosylation at Asn109 | may increase apparent size or contribute to variable migration |
| N-linked glycosylation at Asn112 | may increase apparent size or contribute to variable migration |
| Complex N-linked glycosylation at Asn128 | may increase apparent size or contribute to variable migration |
| N-linked glycosylation at Asn259 | may increase apparent size or contribute to variable migration |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | AZGP1 is secreted | probe conditioned medium alongside lysate |
| Band higher than expected | N-linked glycosylation may affect migration | compare untreated and deglycosylated samples |
| Band lower than expected | signal-peptide cleavage may reduce polypeptide mass | compare lysate and conditioned medium and verify identity with AZGP1 depletion |
| Broad smear instead of sharp band | variable N-linked glycosylation is possible | compare untreated and deglycosylated samples |
| Multiple bands | different glycosylation states or unrelated antibody binding are possible | use AZGP1 depletion to identify specific bands |
| Weak or no signal | a secreted protein may be scarce in the sampled lysate | test conditioned medium and a positive control |
Experiment evidence
Western blot images showing AZGP1 band
M02718-1 · AZGP1
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for AZGP1 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for AZGP1 in Western blot, you can use appendix tissue, which HPA rates highly positive.
Positive control: Appendix (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: Because AZGP1 is secreted, conditioned medium may give a stronger signal than whole-cell lysate.
HPA tissue expression evidence for AZGP1
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 |
|---|---|---|---|---|
| Appendix | endocrine cells | High | Protein (IHC) | HPA → |
| Breast | glandular cells | High | Protein (IHC) | HPA → |
| Duodenum | paneth cells | High | Protein (IHC) | HPA → |
| Prostate | 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 → |
| Adrenal gland | glandular cells | Not detected | Protein (IHC) | HPA → |
| Bone marrow | hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Bronchus | respiratory epithelial cells | Not detected | Protein (IHC) | HPA → |
| Caudate | glial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced AZGP1 Western Blot Tips
Deeper troubleshooting and optimisation questions for AZGP1, answered from its protein features.
How should AZGP1 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 annotated isoforms explain multiple AZGP1 bands?
Isoforms · The supplied record lists one isoform and no alternative sequence. It therefore provides no annotated isoform basis for assigning multiple bands; verify band identity before attributing them to AZGP1.
Which glycosylation sites matter when interpreting AZGP1 bands?
PTM · UniProt lists N-linked sites at positions 109, 112, 128 and 259; positions 109 and 128 are annotated as complex. These are UniProt full-length coordinates, which may differ from numbering used for a processed protein or an antibody.
Does this guide establish induction of AZGP1?
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 AZGP1?
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 M02718-1 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 AZGP1 bands be compared for quantitation?
Quantitation · AZGP1 is secreted and has four annotated N-linked sites. Compare the same sample fraction and the same consistently identified band across samples. The supplied features do not identify an empirical band position or establish that every observed band represents the same AZGP1 form.
Why might AZGP1 migrate differently from its predicted 34.3 kDa?
Interpretation · AZGP1 has a signal peptide at residues 1–20 and four N-linked glycosylation sites. Processing and glycosylation can affect apparent mass, but these features alone do not establish a visible shift or a specific band position.
AZGP1 is secreted and has a signal peptide at residues 1–20. Account for that segment when comparing the 298-residue sequence with a processed protein; the listed features do not give an observed band mass.
Yes. UniProt annotates pyrrolidone carboxylic acid at position 21, immediately after the signal peptide. Position 21 uses full-length UniProt numbering; the annotation alone does not establish a visible band shift.
UniProt lists two disulfide bonds. Comparing reducing and nonreducing samples may help assess whether disulfide-dependent mobility contributes to an unexpected pattern. The record does not establish a particular band pattern or assign any unexpected band to AZGP1.
AZGP1 is secreted and has a signal peptide at residues 1–20. Account for that segment when comparing the 298-residue sequence with a processed protein; the listed features do not give an observed band mass.
Yes. UniProt annotates pyrrolidone carboxylic acid at position 21, immediately after the signal peptide. Position 21 uses full-length UniProt numbering; the annotation alone does not establish a visible band shift.
UniProt lists two disulfide bonds. Comparing reducing and nonreducing samples may help assess whether disulfide-dependent mobility contributes to an unexpected pattern. The record does not establish a particular band pattern or assign any unexpected band to AZGP1.
Boster reagents
AZGP1 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-ZA2G / AZGP1 Rabbit Monoclonal Antibody
Cat # M02718-1
The catalog reports a rabbit monoclonal anti-ZA2G/AZGP1 antibody with reported human reactivity. Its Western blot image shows AZGP1 analysis in HepG2 cell lysate. The supplied evidence does not establish performance in other sample types.
Which to pick: M02718-1 is the only listed AZGP1 antibody. It has a Western blot image from HepG2 cell lysate and reported human reactivity; check suitability separately for other samples.
Source: BosterBio AZGP1 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P25311.
- Human Protein Atlas. AZGP1 tissue expression.
- PMC4053402 — target-verified WB comparison
- PMC4307269 — target-verified WB comparison
- PMC10844717 — target-verified WB comparison