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RBBP7 / Histone-binding protein RBBP7 · Western blot design guide
Design a Western Blot for RBBP7
Source-linked RBBP7 Western blot protocol options, expected-band and isoform facts, troubleshooting for weak or shifted signal, and recommended anti-RBBP7 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
RBBP7 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 | ~47.8 kDa | |
| Gel | 12–15% (standard starting point) | |
| Positive control | Adipose tissue (IHC candidate; verify WB) +4 more | |
| Negative control | Suggested KO / knockdown lysate |
Important caveats
Reasons your observed band may differ from the expected size.
| PTM | Phosphorylated + Acetylated | |
| Caveat | Modification-state controls | |
| Gene-set association | MSigDB Hallmark membership | |
| Isoform | 2 isoform(s) |
Section 1
Source-Linked RBBP7 Western Blot Protocol Options
The A03708 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 A03708) |
| 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 | A03708; 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 RBBP7 Western Blot Band Size?
RBBP7 is predicted at 47.8 kDa; its isoforms and modification sites could affect migration, but no empirical band or visible effect is established.
| Band near 47.8 kDa | Consistent with RBBP7; confirm identity with antibody controls |
| Several bands near the predicted size | Isoforms 1 and 2 are possible contributors, but their migration is unknown |
| Slightly shifted band near 47.8 kDa | RBBP7 has phosphorylation and acetylation sites, but a visible shift is unproven |
| Stronger band in nuclear extract | Consistent with RBBP7's nuclear location |
💡Expected RBBP7 appearanceRBBP7 has a predicted mass of 47.8 kDa, but no empirical band size is supplied; use antibody and nuclear-enrichment controls to assess identity because its modification sites and isoforms do not establish migration.
| Predicted molecular mass | 47.8 kDa is the sequence-based reference size |
| Isoforms 1 and 2 | May differ in size, but their masses and migration are not supplied |
| Phosphorylation at Ser3, Thr10, Ser95 and Ser354 | Could affect migration, but no visible shift is established |
| Lysine acetylation at positions 4, 119 and 159 | Could affect migration, but no visible shift is established |
| Situation | Likely cause | Next action |
|---|---|---|
| No band in lysate | RBBP7 is nuclear, so recovery from the nuclear fraction may be low | Check nuclear extraction and a nuclear loading control |
| Band higher than expected | Its modification sites do not establish the cause of the shift | Check antibody specificity and compare with a validated RBBP7 control |
| Band lower than expected | Isoform masses are unknown, so band identity is uncertain | Confirm with an independent RBBP7 antibody or knockdown |
| Multiple bands | Isoforms 1 and 2 exist, but distinct migration is unproven | Compare bands after RBBP7 knockdown |
| Weak or no signal | Nuclear protein recovery may be low | Enrich the nuclear fraction and verify loading |
Experiment evidence
Western blot images showing RBBP7 band
A03708 · RBBP7
← swipe / scroll for more images →
Real WB data behind the band-size expectations in this section.
Sample controls for RBBP7 Western blot
🧪HPA-IHC candidate guidance (verify in WB): For positive controls for RBBP7 in Western blot, you can use adipose tissue lysate, which has high HPA expression.
Positive control: Adipose tissue (IHC candidate; verify WB)
Negative control: Suggested KO / knockdown lysate
Loading controls: Run GAPDH, β-actin, and a total-protein stain alongside the samples.
⚠️Feasibility: No HPA tissue is listed as not detected, so use siRNA knockdown or a KO line as the negative control.
HPA tissue expression evidence for RBBP7
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 |
|---|---|---|---|---|
| Adipose tissue | adipocytes | High | Protein (IHC) | HPA → |
| Adrenal gland | glandular cells | High | Protein (IHC) | HPA → |
| Appendix | glandular cells | High | Protein (IHC) | HPA → |
| Bone marrow | hematopoietic cells | High | Protein (IHC) | HPA → |
| Breast | adipocytes | High | Protein (IHC) | HPA → |
Lower expression tissues · IHC evidence, not confirmed WB-negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | adipocytes | High | Protein (IHC) | HPA → |
| Adrenal gland | glandular cells | High | Protein (IHC) | HPA → |
| Appendix | glandular cells | High | Protein (IHC) | HPA → |
| Bone marrow | hematopoietic cells | High | Protein (IHC) | HPA → |
| Breast | adipocytes | High | Protein (IHC) | HPA → |
Section 3
Advanced RBBP7 Western Blot Tips
Deeper troubleshooting and optimisation questions for RBBP7, answered from its protein features.
How should RBBP7 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 RBBP7 isoforms produce different bands?
Isoforms · UniProt lists isoforms 1 and 2. In isoform 2, the canonical N-terminal residues 1–6 are replaced by a longer sequence, so the isoforms may differ in mass and recognition by antibodies targeting that region. The entry does not establish whether they resolve as separate bands.
Detection depends on the epitope. Isoform 2 replaces canonical residues 1–6 with a longer sequence, so an antibody requiring those canonical residues may recognize the isoforms differently. Check the antibody’s mapped epitope before comparing isoform signals.
Detection depends on the epitope. Isoform 2 replaces canonical residues 1–6 with a longer sequence, so an antibody requiring those canonical residues may recognize the isoforms differently. Check the antibody’s mapped epitope before comparing isoform signals.
Which phosphorylation annotations matter when interpreting RBBP7 bands?
PTM · The supplied UniProt coordinates list phosphoserine at 3 and 95, phosphothreonine at 10, and a position 354 annotation written as ‘Phosphoserine N-acetylalanine Phosphoserine.’ Keep that position 354 wording as supplied rather than assigning it a single modification. None of these annotations establishes a visible band shift.
The supplied UniProt coordinates list N-acetylalanine at 2 and N6-acetyllysine at 4, 119, and 159; positions 4 and 159 are marked alternate. The position 354 annotation also contains ‘N-acetylalanine’ alongside ‘Phosphoserine’ in the supplied text. Check antibody recognition experimentally if its epitope includes an annotated position.
The supplied UniProt coordinates list N-acetylalanine at 2 and N6-acetyllysine at 4, 119, and 159; positions 4 and 159 are marked alternate. The position 354 annotation also contains ‘N-acetylalanine’ alongside ‘Phosphoserine’ in the supplied text. Check antibody recognition experimentally if its epitope includes an annotated position.
Does this guide establish induction of RBBP7?
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 RBBP7?
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 A03708 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.
What matters when quantifying RBBP7 across samples?
Quantitation · RBBP7 is annotated in the nucleus and has two isoforms. Compare equivalent sample fractions and use an antibody whose recognition of the isoforms is known. If multiple bands appear, establish their identities before combining their signals for quantitation.
Where should RBBP7 migrate relative to its predicted mass?
Interpretation · The supplied predicted mass is 47.8 kDa; no observed band position is available. Use 47.8 kDa as a reference, but do not attribute any difference in apparent mass to a listed modification without experimental evidence.
Compare them with the 47.8 kDa predicted mass and consider the annotated N-terminal isoform difference. The entry also lists phosphorylation and acetylation sites, but their presence alone cannot identify an unexpected band or establish a mobility shift. Use an independent identity check before assigning a band to RBBP7.
Compare them with the 47.8 kDa predicted mass and consider the annotated N-terminal isoform difference. The entry also lists phosphorylation and acetylation sites, but their presence alone cannot identify an unexpected band or establish a mobility shift. Use an independent identity check before assigning a band to RBBP7.
Boster reagents
RBBP7 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-Histone-binding protein RBBP7 RBBP7 Antibody
Cat # A03708
A03708 is the only listed anti-RBBP7 antibody. It is catalogued for human, mouse, and rat and has a Western blot image described as extracts from various cell lines. The supplied caption does not identify the cell lines or provide publication evidence.
Which to pick: A03708 is the only listed option and has a reported Western blot image. Check its listed species reactivity and the image details against your planned sample; the supplied caption does not identify the tested cell lines.
Source: BosterBio RBBP7 gene-info card — filtered to Western-blot-capable antibodies; each card shows that product's actual WB validation figure.
References
- UniProt Consortium. UniProt entry Q16576.
- Human Protein Atlas. RBBP7 tissue expression.
- PMC8270842 — target-verified WB comparison
- PMC10575721 — target-verified WB comparison
- PMC5775914 — target-verified WB comparison