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- Table of Contents
GAS2 / Growth arrest-specific protein 2 · Western blot design guide
Design a Western Blot for GAS2
Real validated GAS2 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-GAS2 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
GAS2 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 | ~34.9 kDa | |
| Observed band | ~35 kDa | |
| Gel | 5–20% (catalog A08589-1) | |
| Positive control | Kidney (IHC candidate; verify WB) +3 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 | 2 isoform(s) |
Section 1
Real Curated GAS2 Western Blot Protocols
The A08589-1 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | human Jurkat, human HepG2, rat liver (catalog A08589-1) |
| Gel % | 5–20% (catalog A08589-1) |
| Load | 30 ug; reducing conditions (catalog A08589-1) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog A08589-1) |
| Membrane | nitrocellulose membrane (catalog A08589-1) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog A08589-1) |
| Primary antibody | A08589-1 · 0.5 μg/mL (catalog A08589-1) |
| Primary incubation | overnight at 4°C (catalog A08589-1) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog A08589-1) |
| Secondary incubation | 1.5 hour at RT (catalog A08589-1) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog A08589-1) |
| Detection | ECL (catalog A08589-1) |
Section 2
What Is the Expected GAS2 Western Blot Band Size?
GAS2 is predicted at 34.9 kDa and observed near 35 kDa; this small difference is consistent with rounding.
| Band near 35 kDa | Matches the reported GAS2 band and its 34.9 kDa predicted mass |
| Additional band at another size | Could reflect isoform 1 or 2; their migration is not established |
| Doublet near 35 kDa | Could reflect phosphorylation at Ser183 or Ser187; a visible shift is not established |
| Weak band in a soluble fraction | May reflect GAS2 association with the cytoskeleton or peripheral membrane |
💡Expected GAS2 appearanceGAS2 is predicted at 34.9 kDa and observed near 35 kDa in whole-cell and tissue lysates; confirm band identity with an appropriate specificity control.
| Predicted molecular mass | 34.9 kDa is consistent with the observed band near 35 kDa |
| Isoforms 1 and 2 | May differ in size, but their individual masses and migration are unspecified |
| Phosphoserine at Ser183 | May affect migration; no visible shift is demonstrated |
| Phosphoserine at Ser187 | May affect migration; no visible shift is demonstrated |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | GAS2 may be poorly recovered from cytoskeletal or peripheral membrane material | Check extraction of insoluble material and include a positive lysate control |
| Band higher than expected | Phosphorylation may affect migration, but the cause is unproven | Compare phosphatase-treated and untreated samples and verify band identity |
| Band lower than expected | The identity of a smaller band is unestablished | Check sample integrity and verify specificity with GAS2 depletion |
| Multiple bands | Isoforms 1 and 2 or phosphorylation may contribute; distinct bands are unproven | Use GAS2 depletion to identify specific bands; compare phosphatase-treated samples if needed |
| Weak or no signal | Recovery may be limited by cytoskeletal or peripheral membrane association | Assess extraction and compare with a positive lysate control |
| Fragments below expected size | Sample proteolysis may produce smaller species | Handle samples with protease inhibitors and verify fragment identity with GAS2 depletion |
Experiment evidence
Western blot images showing GAS2 band
A08589-1 · GAS2
M08589 · GAS2
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for GAS2 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for GAS2 in Western blot, you can use kidney tissue lysate, which HPA rates high for GAS2 expression.
Positive control: Kidney (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: GAS2 is cytoskeletal, so tissue lysates are suitable; HPA reports no detection in adipose tissue.
HPA tissue expression evidence for GAS2
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 |
|---|---|---|---|---|
| Kidney | proximal tubules (cell body) | High | Protein (IHC) | HPA → |
| Duodenum | enterocytes - Microvilli | Medium | Protein (IHC) | HPA → |
| Liver | hepatocytes | Medium | Protein (IHC) | HPA → |
| Testis | elongated or late spermatids | Medium | 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 → |
| Appendix | endocrine 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 GAS2 Western Blot Tips
Deeper troubleshooting and optimisation questions for GAS2, answered from its protein features.
How should GAS2 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.
How might GAS2 isoforms affect band interpretation?
Isoforms · Isoform 2 replaces VLHKQ at canonical positions 137–141 with GFGGQ and lacks canonical residues 142–313. It may produce a smaller band than isoform 1. Check whether the antibody recognizes a region retained in isoform 2 before assigning a band.
Can phosphorylation explain a shifted GAS2 band?
PTM · UniProt lists phosphoserine at canonical positions 183 and 187. These sites fall within the region missing from isoform 2. Their presence does not establish a visible band shift; compare matched samples if testing a phosphorylation-dependent change. State the numbering convention when comparing antibody or paper annotations.
Does this guide establish induction of GAS2?
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 GAS2?
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 A08589-1 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 GAS2 bands be quantified?
Quantitation · Quantify the band assigned to the intended isoform consistently across samples and normalize for loading. Because GAS2 is associated with cytoskeletal and membrane compartments, keep extraction and fraction selection consistent when comparing abundance.
Should GAS2 migrate above its predicted mass?
Interpretation · The canonical protein is predicted at 34.9 kDa, and the observed band is about 35 kDa. These values agree closely. The listed modifications alone do not establish a visible shift.
Growth arrest is a listed GAS2 keyword, but the supplied features give no induction condition or expected change in abundance. Compare matched growth states with a loading control to determine whether signal changes in your samples.
GAS2 is listed in the cytoplasm and cytoskeleton, including stress fibers, and as a peripheral membrane protein. If a whole-cell lysate gives weak signal, examine the relevant fractions and document which fraction was loaded.
First compare bands with the approximately 35 kDa observed band and the shorter sequence of isoform 2. An antibody against canonical residues 142–313 would not detect isoform 2 through that region. The supplied features do not establish the identity of any additional band.
Growth arrest is a listed GAS2 keyword, but the supplied features give no induction condition or expected change in abundance. Compare matched growth states with a loading control to determine whether signal changes in your samples.
GAS2 is listed in the cytoplasm and cytoskeleton, including stress fibers, and as a peripheral membrane protein. If a whole-cell lysate gives weak signal, examine the relevant fractions and document which fraction was loaded.
First compare bands with the approximately 35 kDa observed band and the shorter sequence of isoform 2. An antibody against canonical residues 142–313 would not detect isoform 2 through that region. The supplied features do not establish the identity of any additional band.
Boster reagents
GAS2 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-GAS2 Antibody Picoband®
Cat # A08589-1
Real WB data
Anti-GAS2 Rabbit Monoclonal Antibody
Cat # M08589
Two the supplier anti-GAS2 antibodies have Western blot images. A08589-1 shows an approximately 35 kDa band in the listed human cell, rat liver, and mouse kidney and liver lysates. M08589 shows GAS2 expression in Jurkat lysate; its caption gives no band size or assay conditions.
Which to pick: Choose A08589-1 for its documented human, mouse, and rat lysate examples and reported 35 kDa band. M08589 is listed for human reactivity and has a Jurkat Western blot image, but the supplied caption provides less validation detail.
Source: BosterBio GAS2 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry O43903.
- Human Protein Atlas. GAS2 tissue expression.
- PMC3170402 — target-verified WB comparison
- PMC7008185 — target-verified WB comparison
- PMC6718038 — target-verified WB comparison