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- Table of Contents
BBS1 / BBSome complex member BBS1 · Western blot design guide
Design a Western Blot for BBS1
Source-linked BBS1 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-BBS1 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
BBS1 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 | ~65.1 kDa | |
| Observed band | ~65 kDa | |
| Gel | 5–20% (catalog A03072-2) | |
| 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 | Acetylated | |
| Caveat | — | |
| Gene-set association | MSigDB C7 membership | |
| Isoform | 3 isoform(s) |
Section 1
Source-Linked BBS1 Western Blot Protocol Options
The A03072-2 protocol combines labelled catalog values with standard starting conditions. Published comparisons retain their own sample, reagent and detection scope.
| Sample / lysate | rat kidney, mouse kidney (catalog A03072-2) |
| Gel % | 5–20% (catalog A03072-2) |
| Load | 30 ug; reducing conditions (catalog A03072-2) |
| Transfer | a nitrocellulose membrane at 150 mA for 50-90 minutes (catalog A03072-2) |
| Membrane | nitrocellulose membrane (catalog A03072-2) |
| Blocking | 5% non-fat milk/TBS for 1.5 hour at RT (catalog A03072-2) |
| Primary antibody | A03072-2 · 0.5 μg/mL (catalog A03072-2) |
| Primary incubation | overnight at 4°C (catalog A03072-2) |
| Secondary antibody | goat anti-rabbit IgG-HRP, 1:5000 (catalog A03072-2) |
| Secondary incubation | 1.5 hour at RT (catalog A03072-2) |
| Wash | TBS-0.1%Tween 3 times with 5 minutes each (catalog A03072-2) |
| Detection | ECL (catalog A03072-2) |
Section 2
What Is the Expected BBS1 Western Blot Band Size?
BBS1 is predicted at 65.1 kDa and observed at ~65 kDa; no meaningful difference between these values requires a feature-based explanation.
| Band at ~65 kDa | Matches the reported BBS1 band and its predicted 65.1 kDa mass |
| Additional discrete bands | Could reflect isoforms 1, 2, or 3; confirm their identity |
| Weak ~65 kDa band in whole-cell lysate | BBS1 localization to cilia and centrosomal structures may limit its signal |
| Stronger ~65 kDa band in a cilium-enriched sample | Consistent with BBS1 localization at the cilium membrane |
💡Expected BBS1 appearanceBBS1 is predicted at 65.1 kDa, and a reducing lysate blot reports a ~65 kDa band; confirm identity with appropriate antibody and depletion controls.
| Predicted BBS1 mass | 65.1 kDa, consistent with the reported ~65 kDa band |
| Isoform 1 | May differ in size from other isoforms; its individual mass is unknown |
| Isoform 2 | May differ in size from other isoforms; its individual mass is unknown |
| Isoform 3 | May differ in size from other isoforms; its individual mass is unknown |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | BBS1 may be scarce in the sampled cells or diluted across the whole-cell lysate | Check a positive lysate and consider cilium or centrosome enrichment |
| Band higher than expected | Its identity and reason for slower migration are unestablished | Compare with the ~65 kDa band and test signal loss after BBS1 depletion |
| Band lower than expected | An alternative isoform is possible, but its mass is unknown | Check isoform expression and test signal loss after BBS1 depletion |
| Multiple bands | BBS1 has isoforms 1, 2, and 3, but distinct bands are unconfirmed | Assess isoform expression and use BBS1 depletion to identify specific bands |
| Weak or no signal | The sample may contain little BBS1 at its ciliary or centrosomal locations | Check a positive lysate and enrich the relevant compartment if practical |
Experiment evidence
Western blot images showing BBS1 band
A03072-2 · BBS1
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for BBS1 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for BBS1 in Western blot, you can use adrenal gland tissue, an HPA positive sample with medium 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: BBS1 is ciliary and centrosomal, with medium expression in the listed positive tissues, so lysate signal may be modest.
HPA tissue expression evidence for BBS1
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 | Medium | Protein (IHC) | HPA → |
| Appendix | glandular cells | Medium | Protein (IHC) | HPA → |
| Bronchus | respiratory epithelial cells | Medium | Protein (IHC) | HPA → |
| Epididymis | glandular cells | Medium | Protein (IHC) | HPA → |
| Gallbladder | glandular 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 → |
| Bone marrow | hematopoietic cells | 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 → |
Section 3
Advanced BBS1 Western Blot Tips
Deeper troubleshooting and optimisation questions for BBS1, answered from its protein features.
How should BBS1 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 BBS1 isoforms produce different bands?
Isoforms · Yes. Three isoforms are listed. Isoform 3 replaces residues 1–16 with a longer sequence; isoform 2 lacks residues 242–370 and replaces residues 567–593. These sequence changes may alter apparent band positions, but the features do not establish where each isoform migrates.
Does BBS1 acetylation explain a visible band shift?
PTM · UniProt lists N-acetylalanine at position 2 in the canonical sequence. This modification alone does not establish a visible shift or explain a difference between apparent and predicted mass. The position uses canonical UniProt numbering; isoform 3 replaces the region containing it.
Does this guide establish induction of BBS1?
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 BBS1?
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 A03072-2 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 BBS1 bands be compared during quantitation?
Quantitation · Quantify the same defined band across samples. BBS1 has three isoforms, including an isoform missing residues 242–370, so bands at different positions need separate identification before their signals are combined. Its listed locations include the cilium membrane, cytoplasm and centrosome-associated structures; keep sample preparation consistent.
Does a ~65 kDa band match predicted BBS1 mass?
Interpretation · Yes. The observed ~65 kDa band is close to the 65.1 kDa predicted mass. This agreement supports its assignment but does not establish band identity on its own.
Consider isoform 2, which lacks residues 242–370 and has a replacement at residues 567–593. The listed features do not establish that an observed lower band is isoform 2. Its assignment depends on whether the antibody recognizes sequence retained in that isoform.
BBS1 is listed at the cilium membrane, in the cytoplasm, and at centrosome-associated structures. Differences in how samples collect these compartments could change BBS1 signal. Compare like preparations when interpreting changes in band intensity.
Consider isoform 2, which lacks residues 242–370 and has a replacement at residues 567–593. The listed features do not establish that an observed lower band is isoform 2. Its assignment depends on whether the antibody recognizes sequence retained in that isoform.
BBS1 is listed at the cilium membrane, in the cytoplasm, and at centrosome-associated structures. Differences in how samples collect these compartments could change BBS1 signal. Compare like preparations when interpreting changes in band intensity.
Boster reagents
BBS1 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-BBS1 Antibody Picoband®
Cat # A03072-2
The catalog reports one anti-BBS1 antibody for Western blot: A03072-2, a rabbit polyclonal with reported human, mouse, and rat reactivity. Its supplied WB image shows an approximately 65 kDa band in rat and mouse kidney lysates; no human WB example is provided.
Which to pick: A03072-2 is the only listed option. Its WB example documents rat and mouse kidney lysates under reducing conditions, with primary antibody at 0.5 μg/mL. Human reactivity is listed, but the supplied image does not test a human sample.
Source: BosterBio BBS1 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q8NFJ9.
- Human Protein Atlas. BBS1 tissue expression.
- PMC6507319 — target-verified WB comparison
- PMC10074118 — target-verified WB comparison
- PMC13520375 — target-verified WB comparison