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- Table of Contents
FCGR2B / Low affinity immunoglobulin gamma Fc region receptor II-b · Western blot design guide
Design a Western Blot for FCGR2B
Source-linked FCGR2B Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-FCGR2B 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
FCGR2B 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 kDa | |
| Gel | 12–15% (standard starting point) | |
| Positive control | Placenta (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 | Peptide-blocking control | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 5 isoform(s) |
Section 1
Source-Linked FCGR2B Western Blot Protocol Options
The A01690-1 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | SH-SY5Y cells (catalog A01690-1) |
| Gel % | 12–15% (standard starting point) |
| Load | 20–30 µg total protein per lane; optimize for abundance (standard starting point) |
| Transfer | Short semi-dry transfer; verify retention (standard starting point) |
| Membrane | 0.45 µm PVDF (standard starting point) |
| Blocking | 5% milk or 5% BSA in TBST (standard starting point) |
| Primary antibody | A01690-1 · 1:1000 (catalog A01690-1) |
| 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 FCGR2B Western Blot Band Size?
FCGR2B has a predicted 34 kDa precursor; signal-peptide cleavage, N-linked glycosylation and isoforms may affect migration, but no band position is demonstrated.
| Band near 34 kDa | consistent with the predicted precursor mass; identity requires controls |
| Band above 34 kDa | may reflect occupancy of the three N-linked glycosylation sites |
| Band below 34 kDa | may reflect signal-peptide removal |
| Several bands at different positions | could reflect isoforms or glycosylation states; band identities are unconfirmed |
| Broad smear | could reflect heterogeneous N-linked glycosylation |
💡Expected FCGR2B appearanceUniProt predicts a 34 kDa precursor; signal-peptide cleavage, N-linked glycosylation and isoforms may affect migration, but no empirical band size is supplied, so confirm candidate bands with appropriate identity controls.
| UniProt predicted precursor mass | provides a 34 kDa reference, not a measured band position |
| N-linked glycosylation at Asn106 | may increase apparent size if the site is occupied |
| N-linked glycosylation at Asn180 | may increase apparent size if the site is occupied |
| N-linked glycosylation at Asn187 | may increase apparent size if the site is occupied |
| Signal peptide at residues 1–42 | cleavage can make the mature protein smaller than the precursor |
| Isoforms IIB1, IIB2, IIB3, 4 and 5 | may differ in size; their masses and migration are not supplied |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | poor recovery of this membrane protein | check membrane extraction and use a positive-control lysate |
| Band higher than expected | possible N-linked glycosylation | compare untreated and deglycosylated samples and verify band identity |
| Band lower than expected | possible signal-peptide cleavage | check epitope location and verify band identity |
| Broad smear instead of sharp band | possible variation in N-linked glycosylation | compare untreated and deglycosylated samples |
| Multiple bands | possible isoforms or glycosylation states | compare deglycosylated samples and confirm bands with an independent antibody |
| Weak or no signal | possible poor membrane-protein recovery | check extraction and antibody performance with a positive control |
Experiment evidence
Western blot images showing FCGR2B band
A01690-1 · FCGR2B
A01690-1 · FCGR2B
A01690 · FCGR2B
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for FCGR2B Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for FCGR2B in Western blot, you can use placenta lysate.
Positive control: Placenta (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: As a membrane protein, FCGR2B may be easier to detect in a membrane-enriched fraction.
HPA tissue expression evidence for FCGR2B
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 |
|---|---|---|---|---|
| Placenta | endothelial cells | High | Protein (IHC) | HPA → |
| Bone marrow | hematopoietic cells | Medium | Protein (IHC) | HPA → |
| Lymph node | non-germinal center cells | Medium | Protein (IHC) | HPA → |
| Spleen | cells in white pulp | Medium | Protein (IHC) | HPA → |
| Tonsil | non-germinal center cells | 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 → |
| Breast | adipocytes | Not detected | Protein (IHC) | HPA → |
| Caudate | glial cells | Not detected | Protein (IHC) | HPA → |
| Cerebellum | cells in granular layer | Not detected | Protein (IHC) | HPA → |
| Cerebral cortex | endothelial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced FCGR2B Western Blot Tips
Deeper troubleshooting and optimisation questions for FCGR2B, answered from its protein features.
How should FCGR2B 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.
Which FCGR2B isoforms could produce different bands?
Isoforms · Five isoforms are listed. Relative to the canonical UniProt sequence, IIB3 lacks residues 39..45; isoforms 4 and 5 lack residue 46; IIB2 and 5 lack residues 254..272. Check whether the antibody epitope is retained in each isoform before assigning bands.
How should FCGR2B phosphorylation be assessed?
PTM · UniProt lists phosphotyrosine at position 292, modified by SRC-type Tyr-kinases. Use a site-specific phospho antibody if available and compare its signal with total FCGR2B. Confirm the antibody’s numbering convention before equating its site label with UniProt Tyr292.
Does this guide establish induction of FCGR2B?
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 FCGR2B Western blot?
Transfer · FCGR2B is a single-pass cell-membrane protein with a predicted mass of 34 kDa. Verify recovery and transfer with a total-protein stain, then optimize the method for the observed band. These features do not specify wet or semi-dry transfer.
How should blocking be optimized?
Blocking · Standard workflow guidance: follow the A01690-1 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 FCGR2B be quantified?
Quantitation · Standard workflow guidance: quantify only a validated target band within the linear exposure range. Use consistent sample preparation and loading, retain biological replicates, and avoid interpreting saturation or loading differences as regulation.
Why might FCGR2B migrate differently from its predicted 34 kDa?
Interpretation · The 34 kDa prediction includes the 1..42 signal peptide. FCGR2B also has N-linked glycosylation sites at UniProt positions 106, 180 and 187. Processing or glycosylation could affect mobility, but these features alone do not predict an apparent mass or establish a visible shift.
FCGR2B binds the Fc region of IgG1, IgG3 and IgG4 in immune complexes. If background is suspected, compare a secondary-only control and optimize blocking empirically. The binding feature does not establish that FCGR2B binds detection antibodies under blot conditions.
Normalize FCGR2B signal to total protein and compare the same band or band group across samples. If measuring Tyr292 phosphorylation, report phospho signal relative to total FCGR2B. The listed isoforms and glycosylation sites make band selection important for consistent quantitation.
Check whether bands correspond to listed isoforms, then test whether deglycosylation changes their mobility. The 1..42 signal peptide is another processing consideration. Use an antibody against a second retained epitope to help assess identity; none of these features alone assigns an observed band.
FCGR2B binds the Fc region of IgG1, IgG3 and IgG4 in immune complexes. If background is suspected, compare a secondary-only control and optimize blocking empirically. The binding feature does not establish that FCGR2B binds detection antibodies under blot conditions.
Normalize FCGR2B signal to total protein and compare the same band or band group across samples. If measuring Tyr292 phosphorylation, report phospho signal relative to total FCGR2B. The listed isoforms and glycosylation sites make band selection important for consistent quantitation.
Check whether bands correspond to listed isoforms, then test whether deglycosylation changes their mobility. The 1..42 signal peptide is another processing consideration. Use an antibody against a second retained epitope to help assess identity; none of these features alone assigns an observed band.
Boster reagents
FCGR2B 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-CD32 FCGR2B Antibody
Cat # A01690-1
Real WB data
Anti-CD32-B FCGR2B Antibody
Cat # A01690
Both listed anti-FCGR2B antibodies have Western blot images: A01690-1 with SH-SY5Y cells and A01690 with human placenta extract, each at 1:1000. The supplied images show these sample contexts; they do not demonstrate performance across every listed species.
Which to pick: Choose A01690-1 if you need listed human, mouse, or rat reactivity; its shown blot uses SH-SY5Y cells. Choose A01690 for listed human reactivity and a human placenta blot with 25 µg per lane. Both have WB images.
Source: BosterBio FCGR2B gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry P31994.
- Human Protein Atlas. FCGR2B tissue expression.
- PMC12177957 — target-verified WB comparison
- PMC11034135 — target-verified WB comparison
- PMC12931491 — target-verified WB comparison