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- Table of Contents
CHGB / Secretogranin-1 · Western blot design guide
Design a Western Blot for CHGB
Source-linked CHGB Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-CHGB 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
CHGB 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 | ~78.3 kDa | |
| Observed band | ≈125 kDa | |
| Gel | 8% (catalog M04073) | |
| Positive control | Adrenal gland (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 + Phosphorylated | |
| Caveat | Band identity controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 1 isoform(s) |
Section 1
Source-Linked CHGB Western Blot Protocol Options
The M04073 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human SH-SY5Y, human U251, human A549, human U2OS, rat PC-12, mouse Neuro-2a (catalog M04073) |
| Gel % | 8% (catalog M04073) |
| Load | 30 ug; reducing conditions (catalog M04073) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog M04073) |
| Membrane | nitrocellulose membrane (catalog M04073) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog M04073) |
| Primary antibody | M04073 · 1:500 (catalog M04073) |
| Primary incubation | overnight at 4°C (catalog M04073) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog M04073) |
| Secondary incubation | 1.5 hour at RT (catalog M04073) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog M04073) |
| Detection | ECL (catalog M04073) |
Section 2
What Is the Expected CHGB Western Blot Band Size?
CHGB has a predicted 78.3 kDa precursor and an empirical band near 125 kDa; the cause of the difference is not established.
| Band near 125 kDa | Empirical CHGB-reactive band in reducing whole-cell lysates; confirm identity with controls |
| Band near 78.3 kDa | Possible full-length precursor near its predicted mass |
| Band slightly below the precursor | Possible signal-peptide-cleaved CHGB |
| Diffuse higher-mass signal | Possible variation in chondroitin sulfate modification at Ser93 or Ser239; migration is unproven |
| Weak or absent lysate band | CHGB is secreted and localized to secretory granules |
💡Expected CHGB appearanceUniProt predicts a 78.3 kDa precursor, while antibody QC reports a band near 125 kDa in reducing whole-cell lysates; the cause of the difference is unestablished, so confirm band identity with controls.
| Predicted precursor mass | 78.3 kDa by sequence; the reported CHGB-reactive band is near 125 kDa |
| O-linked chondroitin sulfate site at Ser93 | Modification could affect apparent migration; its contribution is unquantified |
| O-linked chondroitin sulfate site at Ser239 | Modification could affect apparent migration; its contribution is unquantified |
| Signal peptide at residues 1–20 | Cleavage yields a shorter mature protein than the precursor; its apparent mass is unestablished |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | CHGB is secreted and associated with secretory granules | Check a CHGB-expressing positive control and an appropriate secretory-granule or conditioned-medium sample |
| Band higher than expected | The reported band is near 125 kDa, but the reason it exceeds 78.3 kDa is unestablished | Confirm identity by CHGB depletion or an antibody to another epitope |
| Band lower than expected | Signal-peptide cleavage or processing at paired basic residues may produce smaller forms | Check antibody epitope coverage and compare with a CHGB-positive control |
| Broad smear instead of sharp band | Variable chondroitin sulfate modification at Ser93 or Ser239 is possible | Compare treated and untreated samples and confirm CHGB identity with an independent antibody |
| Multiple bands | CHGB processing at paired basic residues is possible | Check antibody epitope coverage and compare bands with a CHGB-depleted sample |
| Weak or no signal | CHGB may be scarce in the sampled lysate because it is secreted | Test a CHGB-positive control and enrich secretory-granule or conditioned-medium material |
Experiment evidence
Western blot images showing CHGB band
M04073 · CHGB
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for CHGB Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for CHGB in Western blot, you can use adrenal gland tissue, which HPA rates highly positive.
Positive control: Adrenal gland (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: CHGB is secreted, so signal in tissue lysate may be lower after secretion.
HPA tissue expression evidence for CHGB
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 |
|---|---|---|---|---|
| Adrenal gland | glandular cells | High | Protein (IHC) | HPA → |
| Appendix | endocrine cells | High | Protein (IHC) | HPA → |
| Cerebellum | synaptic glomeruli - core | High | Protein (IHC) | HPA → |
| Colon | endocrine cells | High | Protein (IHC) | HPA → |
| Duodenum | endocrine 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 → |
| Bone marrow | hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Breast | adipocytes | 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 CHGB Western Blot Tips
Deeper troubleshooting and optimisation questions for CHGB, answered from its protein features.
How should CHGB 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 CHGB isoforms explain multiple bands?
Isoforms · Only one isoform is supplied, with no alternative sequence. The supplied features therefore do not support assigning distinct bands to annotated isoforms. Check antibody recognition and the protein's listed modifications and cleavage-related features before interpreting additional bands.
Which glycosylation sites matter when interpreting CHGB bands?
PTM · UniProt lists O-linked chondroitin sulfate at Ser93 and Ser239. Use these UniProt sequence coordinates when comparing site-specific results; antibody or paper numbering may differ. The annotations alone do not predict a particular band shift.
UniProt lists 24 phosphorylated residues, including Ser130, Ser225, Ser367, Ser377 and Ser380 annotated as phosphorylated by FAM20C. Use UniProt sequence coordinates when choosing site-specific assays; other numbering conventions may differ. A listed site does not establish that phosphorylation caused a visible band shift.
UniProt lists 24 phosphorylated residues, including Ser130, Ser225, Ser367, Ser377 and Ser380 annotated as phosphorylated by FAM20C. Use UniProt sequence coordinates when choosing site-specific assays; other numbering conventions may differ. A listed site does not establish that phosphorylation caused a visible band shift.
Does this guide establish induction of CHGB?
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 CHGB?
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 M04073 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.
What should be controlled when quantifying CHGB Western blots?
Quantitation · Define which CHGB-reactive band or bands you will quantify and apply that choice consistently. The approximately 125 kDa observed band differs from the 78.3 kDa predicted mass, and the supplied features include glycosylation and other modifications. Band position alone cannot establish which molecular form each signal represents.
Why can CHGB appear near 125 kDa instead of 78.3 kDa?
Interpretation · The observed band is approximately 125 kDa, while the predicted mass is 78.3 kDa. CHGB has O-linked chondroitin sulfate sites at Ser93 and Ser239 and many other annotated modifications. These features warrant checking modified CHGB, but their presence alone does not establish the cause or size of the apparent mass difference.
CHGB has a 1–20 signal peptide and a keyword for cleavage on pairs of basic residues. Processing is therefore relevant when investigating smaller bands, but no cleavage products or cleavage positions are supplied. Do not assign a smaller band to a specific fragment from these features alone.
CHGB is annotated as secreted and located in neuroendocrine and endocrine secretory granules. Consider both cellular material and secreted material when deciding where to look for a CHGB signal, according to the sample system being studied.
CHGB has a 1–20 signal peptide and a keyword for cleavage on pairs of basic residues. Processing is therefore relevant when investigating smaller bands, but no cleavage products or cleavage positions are supplied. Do not assign a smaller band to a specific fragment from these features alone.
CHGB is annotated as secreted and located in neuroendocrine and endocrine secretory granules. Consider both cellular material and secreted material when deciding where to look for a CHGB signal, according to the sample system being studied.
Boster reagents
CHGB 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-Chromogranin B CHGB Monoclonal Antibody
Cat # M04073
The catalog reports one anti-CHGB antibody, M04073, with a Western blot image from human SH-SY5Y, U251, A549 and U2OS, rat PC-12, and mouse Neuro-2a lysates. The reported band is approximately 125 kDa versus an expected 78 kDa. No publication evidence is supplied.
Which to pick: M04073 is the only listed option. Its stated reactivity is human, mouse, and rat; its WB caption documents those specific cell lysates under reducing conditions at 1:500. Account for the difference between the observed 125 kDa band and expected 78 kDa size.
Source: BosterBio CHGB gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P05060.
- Human Protein Atlas. CHGB tissue expression.
- PMC7526459 — target-verified WB comparison
- PMC6238609 — target-verified WB comparison
- PMC2996542 — target-verified WB comparison