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- Table of Contents
ADRB2 / Beta-2 adrenergic receptor · Western blot design guide
Design a Western Blot for ADRB2
Real validated ADRB2 Western blot protocols, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-ADRB2 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
ADRB2 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 | ~46.5 kDa | |
| Observed band | 47 kDa | |
| Gel | 12–15% (standard starting point) | |
| Negative control | Suggested KO / knockdown lysate |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Glycosylated + Phosphorylated | |
| Caveat | Modification-state controls | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 1 isoform(s) |
Section 1
Real Curated ADRB2 Western Blot Protocols
The PB9365 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | Target-positive lysate and matched negative control (standard starting point) |
| 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 | PB9365; 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 ADRB2 Western Blot Band Size?
ADRB2 is predicted at 46.5 kDa and observed at 47 kDa; the cause of the small difference is not established.
| Band near 47 kDa | Matches the empirical ADRB2 band; confirm identity with antibody controls. |
| Band above 47 kDa | N-linked glycosylation at Asn6 or Asn15 could affect migration; identity needs confirmation. |
| Doublet near 47 kDa | Different glycosylation states are possible, but distinct bands are not established. |
| Lower band after deglycosylation | Loss of N-linked glycans could change migration; the size change is not established. |
💡Expected ADRB2 appearanceADRB2 has a predicted mass of 46.5 kDa and an empirical band at 47 kDa; confirm band identity with ordinary antibody controls because the supplied features do not establish the cause of migration.
| Predicted molecular mass | Places the 413-residue receptor at 46.5 kDa; the observed band is 47 kDa. |
| Asn6 N-linked glycosylation | Could alter apparent size; no shift is quantified. |
| Asn15 N-linked glycosylation | Could alter apparent size; no shift is quantified. |
| N-linked glycosylation at Asn6 and Asn15 | Could influence native migration; the contribution to the 47 kDa band is unknown. |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | A membrane receptor may be poorly recovered from the lysate. | Check membrane extraction and include a positive control. |
| Band higher than expected | N-linked glycosylation could affect migration. | Compare treated and untreated samples and confirm band identity. |
| Band lower than expected | Glycosylation state could differ, but the cause is unestablished. | Check antibody specificity and compare glycosylation controls. |
| Broad smear instead of sharp band | Heterogeneous N-linked glycosylation is possible. | Compare treated and untreated samples and check loading. |
| Multiple bands | Different glycosylation states are possible; distinct species are not established. | Confirm specificity with an independent antibody or target depletion. |
| Weak or no signal | Membrane protein recovery or antibody detection may be inadequate. | Check extraction, loading, and a positive control. |
Experiment evidence
Western blot images showing ADRB2 band
PB9365 · ADRB2
M00072 · ADRB2
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for ADRB2 Western blot
🧪For positive controls for ADRB2 in Western blot, you can use an HPA-IHC candidate positive sample once one is identified; none is supplied here.
Positive control: No high/medium HPA tissue identified
Negative control: Suggested KO / knockdown lysate
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside.
⚠️Feasibility: ADRB2 is a multi-pass membrane protein, and the supplied HPA data cannot establish tissue controls.
HPA tissue expression evidence for ADRB2
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 |
|---|---|---|---|---|
| No high/medium HPA tissues identified in the supplied evidence. | ||||
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| No lower-expression tissue rows available in the supplied evidence. | ||||
Section 3
Advanced ADRB2 Western Blot Tips
Deeper troubleshooting and optimisation questions for ADRB2, answered from its protein features.
How should ADRB2 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.
Do listed isoforms explain multiple ADRB2 bands?
Isoforms · UniProt lists one isoform and no alternative sequence. Do not assign additional bands to a documented ADRB2 isoform on this evidence alone.
Could glycosylation affect ADRB2 migration?
PTM · UniProt lists N-linked glycosylation at Asn6 and Asn15. Compare samples with consistent preparation when assessing migration; these sites alone do not establish how far a band will shift. Positions here use UniProt numbering, which may differ from paper or antibody numbering.
UniProt lists Tyr141, Ser246, PKA sites Ser261, Ser262, Ser345 and Ser346, and BARK sites Ser355 and Ser356. Record the conditions compared and use UniProt coordinates when matching site-specific antibodies; paper or antibody numbering may differ. Listed sites alone do not establish a visible band shift.
UniProt lists Tyr141, Ser246, PKA sites Ser261, Ser262, Ser345 and Ser346, and BARK sites Ser355 and Ser356. Record the conditions compared and use UniProt coordinates when matching site-specific antibodies; paper or antibody numbering may differ. Listed sites alone do not establish a visible band shift.
Does this guide establish induction of ADRB2?
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.
What transfer method to use for ADRB2 Western blot?
Transfer · ADRB2 is a multi-pass membrane protein. Choose and validate a transfer method for membrane proteins by checking whether ADRB2 remains in the gel and whether the expected region reaches the membrane. The supplied features do not specify a uniquely optimal method.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the PB9365 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 ADRB2 bands be quantified?
Quantitation · ADRB2 is found at the cell membrane and Golgi apparatus. Keep membrane recovery and sample preparation consistent across conditions, and quantify the same defined band region within a linear signal range. Interpret changes cautiously if the band pattern or recovery differs.
Why is the ADRB2 band near 47 kDa?
Interpretation · The observed band is 47 kDa, close to the predicted 46.5 kDa. ADRB2 has listed modifications, but their presence alone does not establish a visible shift or explain the apparent mass.
Check the 47 kDa observed region first. ADRB2 has N-linked sites at Asn6 and Asn15, phosphorylation sites, and 4-hydroxyproline at Pro382 and Pro395, but these features alone cannot identify an extra band. The supplied record lists one isoform, so confirm any additional band independently before assigning its cause.
Check the 47 kDa observed region first. ADRB2 has N-linked sites at Asn6 and Asn15, phosphorylation sites, and 4-hydroxyproline at Pro382 and Pro395, but these features alone cannot identify an extra band. The supplied record lists one isoform, so confirm any additional band independently before assigning its cause.
Boster reagents
ADRB2 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-beta 2 Adrenergic Receptor/ADRB2 Antibody Picoband®
Cat # PB9365
Real WB data
Anti-beta 2 Adrenergic Receptor ADRB2 Rabbit Monoclonal Antibody
Cat # M00072
Both listed anti-ADRB2 antibodies have WB images. PB9365 shows a 47 kDa band in rat brain tissue lysate at 0.5 µg/mL; M00072 shows ADRB2 analysis in A431 cell lysate. The supplied captions document these sample contexts only.
Which to pick: For rat brain tissue, PB9365 has a matching WB image. For A431 cell lysate, M00072 has a WB image. PB9365 lists human and rat reactivity; M00072 lists human, mouse, and rat reactivity.
Source: BosterBio ADRB2 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P07550.
- Human Protein Atlas. ADRB2 tissue expression.
- PMC11617210 — target-verified WB comparison
- PMC12343932 — target-verified WB comparison
- PMC12219006 — target-verified WB comparison