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- Table of Contents
SCG2 / Secretogranin-2 · Western blot design guide
Design a Western Blot for SCG2
Source-linked SCG2 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-SCG2 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
SCG2 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 | ~70.9 kDa | |
| Gel | 12–15% (standard starting point) | |
| 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 | Phosphorylated + Cleaved | |
| Caveat | Processing-state controls | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 1 isoform(s) |
Section 1
Source-Linked SCG2 Western Blot Protocol Options
The A04779 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | extracts of various cell lines, (catalog A04779) |
| 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 | Blocking buffer: 3% nonfat dry milk in TBST (catalog A04779) |
| Primary antibody | A04779 · 1:1000 (catalog A04779) |
| Primary incubation | Overnight at 4 °C (standard starting point) |
| Secondary antibody | Goat Anti-Rabbit IgG, 1:10000 (catalog A04779) |
| Secondary incubation | 1 h at room temperature (standard starting point) |
| Wash | 3 × 5 min in TBST (standard starting point) |
| Detection | ECL (catalog A04779) |
Section 2
What Is the Expected SCG2 Western Blot Band Size?
SCG2 has a predicted 70.9 kDa precursor; signal-peptide, propeptide, and basic-residue cleavage could yield smaller forms, but their migration has not been demonstrated here.
| Band near 70.9 kDa | May represent the full-length SCG2 precursor; identity requires confirmation |
| Band below 70.9 kDa | May reflect signal-peptide or propeptide removal |
| Fragments below the precursor band | May reflect cleavage at paired basic residues |
| Multiple bands | May reflect precursor and processed SCG2 forms |
| Little or no band in whole-cell lysate | SCG2 is secreted into neuroendocrine and endocrine granules |
💡Expected SCG2 appearanceUniProt predicts a 70.9 kDa SCG2 precursor; cleavage may produce smaller forms, but no empirical band size is supplied, so confirm band identity with appropriate controls.
| Predicted full-length precursor mass | Provides a 70.9 kDa sequence-based reference, not a measured migration |
| Signal peptide at residues 1–27 | Its removal makes the mature chain smaller than the precursor |
| Propeptide at residues 28–30 | Its removal further reduces the mature chain's size |
| Signal-peptide and propeptide processing | Together, these cleavages make the processed chain smaller than the 70.9 kDa precursor; its apparent mass is unknown |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | SCG2 is secreted and localized to secretory granules | Check conditioned medium or a granule-enriched sample alongside a positive control |
| Band lower than expected | Signal-peptide or propeptide removal may lower precursor size | Compare precursor and processed samples and verify antibody recognition |
| Multiple bands | Precursor and processed SCG2 forms may coexist | Compare samples with different processing states and verify each band's identity |
| Weak or no signal | SCG2 abundance may be low in the sampled cell fraction after secretion | Check a known SCG2-positive sample and conditioned medium |
| Fragments below expected size | Cleavage at paired basic residues may generate smaller SCG2 products | Check the antibody epitope and compare fragment patterns with a positive control |
Experiment evidence
Western blot images showing SCG2 band
A04779 · SCG2
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for SCG2 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for SCG2 in Western blot, you can use adrenal gland tissue, which HPA scores as high expression.
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 alongside the samples.
⚠️Feasibility: Because SCG2 is secreted, whole-cell lysates may give weak signal; consider conditioned medium.
HPA tissue expression evidence for SCG2
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 | medullary cells | High | Protein (IHC) | HPA → |
| Colon | endocrine cells | High | Protein (IHC) | HPA → |
| Duodenum | endocrine cells | High | Protein (IHC) | HPA → |
| Hypothalamus | neuronal projections | High | Protein (IHC) | HPA → |
| Pituitary gland | cells in anterior | 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 → |
| Appendix | endocrine cells | 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 → |
Section 3
Advanced SCG2 Western Blot Tips
Deeper troubleshooting and optimisation questions for SCG2, answered from its protein features.
How should SCG2 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.
Are alternative SCG2 isoforms listed?
Isoforms · The supplied features list one isoform and no alternative sequence. Do not assign distinct bands to annotated isoforms on this evidence alone; assess processing and antibody recognition when interpreting multiple bands.
Which SCG2 modifications should I consider when interpreting bands?
PTM · UniProt lists sulfotyrosine at position 151 and phosphoserines at 174, 268, 432, 532, 555, and 556. These are UniProt sequence coordinates; paper or antibody numbering may differ. Their presence does not establish a visible band shift or explain any difference from 70.9 kDa.
The supplied record includes a glycoprotein keyword but lists no glycosylation sites. It does not establish whether glycosylation causes a band shift. Avoid assigning a shifted band to a particular glycan from these features alone.
The supplied record includes a glycoprotein keyword but lists no glycosylation sites. It does not establish whether glycosylation causes a band shift. Avoid assigning a shifted band to a particular glycan from these features alone.
Does this guide establish induction of SCG2?
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 SCG2?
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 A04779 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 multiple SCG2 bands?
Quantitation · Quantify each reproducible band separately until its identity is established. SCG2 has annotated signal and propeptide regions and a cleavage keyword, so bands may represent different processed forms. Record the sample fraction and use the same band definition across samples.
Should SCG2 migrate at its predicted 70.9 kDa?
Interpretation · 70.9 kDa is the predicted mass, and no observed band is supplied. SCG2 has a signal peptide at residues 1–27 and a propeptide at 28–30, so processing can change the protein present in a sample. These features alone do not establish its apparent Western blot mass or a visible shift.
SCG2 is annotated with cleavage on pairs of basic residues, but specific cleavage products and their masses are not supplied. Its signal peptide spans residues 1–27 and its propeptide spans 28–30. Consider processing when assessing smaller bands, and confirm which SCG2 region the antibody recognizes.
SCG2 is secreted and localized to neuroendocrine and endocrine secretory granules. Consider both cellular and secreted material when selecting samples, and keep the sampled fraction consistent across comparisons.
SCG2 is annotated with cleavage on pairs of basic residues, but specific cleavage products and their masses are not supplied. Its signal peptide spans residues 1–27 and its propeptide spans 28–30. Consider processing when assessing smaller bands, and confirm which SCG2 region the antibody recognizes.
SCG2 is secreted and localized to neuroendocrine and endocrine secretory granules. Consider both cellular and secreted material when selecting samples, and keep the sampled fraction consistent across comparisons.
Boster reagents
SCG2 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-Secretogranin-2 SCG2 Antibody
Cat # A04779
The catalog reports one anti-SCG2 antibody, A04779, with reported Human and Rat reactivity. Its Western blot image is described as using various cell line extracts at 1:1000 dilution and 25 µg per lane; the caption does not identify the cell lines or their species.
Which to pick: A04779 is the only listed option and has a Western blot image. Check whether its reported Human or Rat reactivity fits your sample; the image caption does not establish which species were tested.
Source: BosterBio SCG2 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P13521.
- Human Protein Atlas. SCG2 tissue expression.
- PMC8436928 — target-verified WB comparison
- PMC13463045 — target-verified WB comparison
- PMC10551594 — target-verified WB comparison