RNF7 / RING-box protein 2 · IHC design guide

Design Immunohistochemistry for RNF7

Plan chromogenic RNF7 IHC in paraffin sections using the IHC-validated antibody M05620 at 1:25 (datasheet M05620). Compare staining in kidney tubule cells, bone marrow hematopoietic cells and splenic red pulp cells, while accounting for the reported low consistency between antibody staining and RNA expression (HPA tissue IHC).

Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use

Immunohistochemistry protocol sheet for RNF7 (IHC for RNF7): expected localisation Cytoplasmic staining in several tissues (HPA tissue IHC), antibody M05620, validated IHC image, and IHC protocol steps — Printable **RNF7** IHC protocol sheet — expected localisation Cytoplasmic staining in several tissues (HPA tissue IHC), antibody M05620, controls and protocol steps. Open the full RNF7 IHC guide →

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RNF7 Immunohistochemistry Experimental Design Guide

Expected localisation, validated protocols, controls and antibodies — the at-a-glance facts below, then the full design guide.

Must know before staining

	Expected localisation	Cytoplasmic staining in several tissues (HPA tissue IHC)
	Staining pattern	Cytoplasmic staining across several tissue cell types (HPA tissue IHC)
	Antigen retrieval	Citrate pH 6 HIER, heat-mediated (datasheet M05620)
	Positive control ⓘ	Bone marrow+4 more · see all
	Negative control ⓘ	Bronchus+4 more · see all

Important caveats

Reasons your staining may differ from the expected pattern.

	Fixation	Use formaldehyde-fixed paraffin sections (selected-SKU IHC image M05620); keep fixation consistent (standard IHC practice; not target-specific)
	Caveat	Antibody staining has low consistency with RNA expression (HPA tissue IHC)
	Regulation	Expression varies by tissue (UniProt)
	Isoform / epitope	Four isoforms; epitope coverage is unspecified (UniProt; datasheet M05620)

Section 1

Recommended RNF7 IHC & IF Protocols

The catalog antibody’s IHC-P protocol is paired with one published RNF7 protocol for paraffin sections (PMC9107170).

Recommended immunohistochemistry (IHC-P) protocol parameters
Sample	Formaldehyde-fixed, paraffin-embedded human kidney tissue (datasheet M05620)
Fixation	Image formalin-fixed; duration unreported (datasheet M05620); verify before use.
Sectioning	4–5 µm sections on charged slides (standard)
Deparaffinisation	Xylene, graded ethanol series to water (standard)
Antigen retrieval	Heat retrieval: Citrate pH 6 (datasheet M05620); 20 min, 95–100 °C (standard)
Peroxidase block	3% H2O2, 10 min, room temperature (standard)
Blocking	10% normal serum of the secondary host, 30 min, room temperature (standard)
Primary antibody	Rabbit anti-RNF7, 1:25 (datasheet M05620)
Primary incubation	1 hours at 37°C (datasheet M05620)
Detection	HRP-polymer secondary, DAB chromogen 5–10 min (standard)
Counterstain	Hematoxylin, blue, dehydrate and mount (standard)
Expected result	RNF7-positive staining in hematopoietic cells of bone marrow (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression in several tissues. No signal in the no-primary control.

💡Decision noteStart with heat-mediated citrate pH 6 retrieval (datasheet M05620); the published paraffin-section protocol also uses citric acid buffer at pH 6.0 (PMC9107170).

Section 2

What Is the Expected RNF7 Staining Pattern?

In paraffin-section IHC, expect RNF7 staining mainly in the cytoplasm of cells within several tissues, particularly bone-marrow hematopoietic cells, kidney tubular cells and splenic red-pulp cells (HPA tissue IHC: High). RNF7 has no transmembrane segment and is annotated in both cytoplasm and nucleus (UniProt Q9UBF6). Treat the tissue pattern as a guide: the HPA IHC antibody is Approved, but its staining has low consistency with RNA expression (HPA tissue IHC).

What am I looking at on my slide?
Clear cytoplasmic staining in hematopoietic cells, kidney tubular cells or splenic red-pulp cells.	This matches the reported high-staining cell populations and cytoplasmic tissue profile (HPA tissue IHC: High; cytoplasmic expression). Judge the named cells rather than the overall darkness of a mixed tissue section (general IHC practice).
Staining appears predominantly at cell borders or outside cells, with little identifiable cytoplasmic signal.	A membrane-only or extracellular pattern does not match the reported tissue-IHC profile (HPA tissue IHC: cytoplasmic expression). Review morphology and detection controls before calling it RNF7-specific (general IHC practice).
Strong staining occurs mainly in a cell population reported as not detected, such as lung alveolar cells.	This conflicts with that HPA tissue observation (HPA tissue IHC: alveolar cells Not detected). Consider antibody cross-reactivity or endogenous detection activity, then check cell identity and controls (general IHC practice).
Color spreads across the section or obscures cell boundaries and the counterstain.	Diffuse background prevents a reliable compartment or cell-type call (general IHC practice). It is not the discrete cytoplasmic pattern reported across several tissues (HPA tissue IHC: cytoplasmic expression).
No signal appears in hematopoietic cells of bone marrow or tubular cells of kidney.	An assay failure is possible because both are reported high-staining populations (HPA tissue IHC: High). Check tissue preservation, staining controls and detection performance before interpreting absence as biological loss (general IHC practice).

💡Expected RNF7 appearanceCall an IHC result positive when identifiable bone-marrow hematopoietic cells, kidney tubular cells or splenic red-pulp cells show clear cytoplasmic staining at the reported high level (HPA tissue IHC: High; cytoplasmic expression); diffuse, border-only or wrong-cell staining needs control-based review (general IHC practice).

How each factor affects the staining
IHC tissue pattern	The strongest listed staining is in bone-marrow hematopoietic cells, kidney tubular cells and splenic red-pulp cells (HPA tissue IHC: High). Several listed glandular and neuronal populations stain at medium levels (HPA tissue IHC: Medium), so intensity depends on the cells examined.
Antibody evidence	The listed antibody, HPA036995, is Approved for IHC; the tissue record also reports low consistency between antibody staining and RNA expression (HPA antibodies: IHC Approved; HPA tissue IHC: reliability description). Treat an unexpected result as requiring controls.
Subcellular interpretation	UniProt places RNF7 in cytoplasm and nucleus, while HPA tissue IHC describes cytoplasmic expression (UniProt Q9UBF6: subcellular location; HPA tissue IHC: profile). Nuclear staining alone therefore needs assessment against the IHC tissue pattern; it is not settled by the location annotation.
Protein form and epitope	RNF7 has 4 listed isoforms and a chain spanning residues 2–113, with no signal peptide, propeptide or transmembrane segment (UniProt Q9UBF6: isoforms, processing, topology). Without an antibody epitope map, these facts cannot predict isoform coverage or staining strength.
Modified residues	UniProt lists N-acetylalanine at residue 2 and CK2-dependent phosphothreonine at residue 10 (UniProt Q9UBF6: modified residues). Their effects on this antibody's IHC signal are unreported; do not infer a retrieval requirement or fixation sensitivity from them.
IF/ICC Q: Should the IHC cytoplasmic pattern also be expected in IF/ICC?	A: HPA ICC-IF reports mainly nucleoplasmic localization with additional cytosolic signal (HPA subcellular: supported locations). Interpret IF/ICC using its own localization evidence; the paraffin-section IHC profile describes cytoplasmic staining in tissues (HPA tissue IHC).

Why is my staining missing, weak or wrong?
Situation	Likely cause	Next action
A high-staining reference population is blank.	The run may have failed, or the selected section may lack the expected cells (general IHC practice); bone-marrow hematopoietic and kidney tubular cells are reported High (HPA tissue IHC).	Confirm cell identity and section quality, then review the run's positive control, primary-antibody step and detection reagents (general IHC practice). Do not score biological absence until the run works.
A reported negative cell population stains strongly.	Cross-reactivity or endogenous detection activity may contribute (general IHC practice); lung alveolar cells and heart cardiomyocytes are reported Not detected (HPA tissue IHC).	Check a no-primary control and the cells actually stained; assess endogenous enzyme blocking for chromogenic detection (general IHC practice). Compare the result with HPA at the cell-type level.
The whole section has diffuse brown background.	Excess detection signal or inadequate blocking can obscure cell-specific staining (general IHC practice). Diffuse signal cannot establish the cytoplasmic profile reported for RNF7 (HPA tissue IHC).	Review no-primary controls, blocking, reagent concentration, washes and chromogen development time; adjust one general IHC workflow variable at a time (general IHC practice).
Signal is confined to cell borders or extracellular material.	This location conflicts with the reported cytoplasmic tissue pattern and RNF7's lack of a transmembrane segment (HPA tissue IHC: profile; UniProt Q9UBF6: topology).	Recheck morphology and background controls, then seek independent antibody evidence before assigning RNF7 specificity (general IHC practice). Do not score border-only color as cytoplasmic positivity.
Nuclear staining dominates a paraffin-section IHC result.	RNF7 has a nuclear annotation and nucleoplasmic ICC-IF signal (UniProt Q9UBF6; HPA subcellular), but the supplied tissue-IHC profile describes cytoplasmic expression (HPA tissue IHC).	Record nuclear and cytoplasmic scores separately, check controls, and compare like-for-like IHC tissue images before deciding whether the pattern is credible (general IHC practice).
Cardiomyocytes stain despite the HPA IHC negative listing.	UniProt reports RNF7 expression in heart, while HPA reports cardiomyocytes Not detected by tissue IHC (UniProt Q9UBF6: tissue specificity; HPA tissue IHC: heart muscle). These observations use different evidence types.	Confirm the stained cells and run controls, then report the discrepancy as assay-specific evidence rather than treating either source as a guaranteed IHC result (general IHC practice).

Source: UniProt Q9UBF6 Human Protein Atlas tissue Human Protein Atlas subcellular

Experiment evidence

IHC & IF images showing RNF7 staining

M05620 staining RNF7 in human kidney tissue sections by Immunohistochemistry (IHC-P -paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 3% BSA for 0. 5 hour at room temperature; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody (1/25) for 1 hours at 37°C. A undiluted biotinylated goat polyvalent antibody was used as the secondary antibody.

M05620 · IHCM05620 staining RNF7 in human kidney tissue sections by Immunohistochemistry (IHC-P -paraformaldehyde-fixed, paraffin-embedded sections). Ti

HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000114125-RNF7/tissue/cerebral+cortex#imid_8978330

HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000114125-RNF7/tissue/colon#imid_8965368

HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000114125-RNF7/tissue/liver#imid_8965353

HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000114125-RNF7/tissue/kidney#imid_8965362

HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA036995; green = RNF7, blue = nucleus, red = microtubules.

← swipe / scroll for more images →

Real IHC / IF data behind the staining-pattern expectations in this section.

Sample controls for RNF7 IHC & IF

🧪Run kidney first and require staining in tubular cells (HPA: High in kidney tubular cells); use bronchus respiratory epithelial cells as the negative tissue (HPA: Not detected). On the kidney slide, intact counterstained cells without specific signal can show local background, but the supplied HPA rows do not establish a kidney cell type as RNF7-negative (HPA: kidney tubular cells High).

Positive control tissue: Bone marrow (Hematopoietic cells, HPA High)

Negative control tissue: Bronchus (HPA Not detected)

ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show RNF7 in A-431, U-251MG, U2OS, KOLF2.1J, with annotated localisation: Nucleoplasm (supported) (HPA subcellular).

Technical controls: Include a no-primary secondary-only control, a host-species- and isotype-matched control for a monoclonal primary or matched nonimmune IgG for a polyclonal primary, and RNF7-knockout material as a biological negative if available (standard IHC practice). Block endogenous peroxidase and check endogenous biotin background in kidney when using the caption’s biotinylated secondary antibody (catalog caption: biotinylated secondary; standard IHC practice).

⚠️Feasibility: No target-specific fixation window or fixation effect is reported in the supplied evidence, and the selected M05620 tissue-IHC caption’s fixative is unreported (M05620 caption: fixative unreported). The caption uses heat retrieval in citrate buffer at pH 6, but does not establish whether RNF7 staining depends on retrieval (M05620 caption: citrate heat retrieval). HPA has ICC-IF images in A-431, U-251MG, U2OS and KOLF2.1J, but the supplied evidence does not show that frozen sections or IF are easier than paraffin IHC; kidney staining with a biotinylated detection system warrants an endogenous biotin check (HPA: ICC-IF image cell lines; M05620 caption: biotinylated secondary; standard IHC practice).

HPA tissue IHC evidence for RNF7

Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — Low consistency between antibody staining and RNA expression data.). Rows are taken directly from the HPA tissue chart — click any row's HPA link to view the source.

Positive expression · recommended positive controls

Tissue	Cell type	Level	Evidence	Source
Bone marrow	Hematopoietic cells	High	Protein (IHC)	HPA →
Kidney	Cells in tubules	High	Protein (IHC)	HPA →
Spleen	Cells in red pulp	High	Protein (IHC)	HPA →
Adrenal gland	Glandular cells	Medium	Protein (IHC)	HPA →
Appendix	Glandular cells	Medium	Protein (IHC)	HPA →

Undetected expression · recommended negative controls

Tissue	Cell type	Level	Evidence	Source
Bronchus	Respiratory epithelial cells	Not detected	Protein (IHC)	HPA →
Esophagus	Squamous epithelial cells	Not detected	Protein (IHC)	HPA →
Heart muscle	Cardiomyocytes	Not detected	Protein (IHC)	HPA →
Hippocampus	Glial cells	Not detected	Protein (IHC)	HPA →
Lung	Alveolar cells	Not detected	Protein (IHC)	HPA →

Section 3

Advanced RNF7 IHC Tips

Use matched controls and cell-level scoring to troubleshoot RNF7 staining in paraffin sections; interpret patterns alongside documented tissue and subcellular evidence.

Which retrieval condition should I try first for weak RNF7 staining in paraffin sections?

Start with heat-mediated antigen retrieval in citrate buffer at pH 6 (datasheet M05620). The selected IHC image used this retrieval before a 1:25 primary antibody incubation for 1 hour at 37°C (M05620 tissue-IHC caption). If staining remains weak, compare retrieval duration on adjacent sections while holding antibody dilution, detection and chromogen development constant; include a no-primary control to reveal background from the detection system (standard IHC practice). Record whether the change improves staining in kidney tubular cells, a reported high-staining population, without increasing diffuse staining elsewhere (HPA: High in kidney tubular cells).

How should I assess whether fixation is masking RNF7 staining?

Target-specific fixation sensitivity is unknown from the supplied evidence, so treat fixation as an experimental variable rather than an established RNF7 effect. Compare sections processed with documented fixation conditions, using the same citrate retrieval at pH 6, primary antibody dilution and chromogen development for each section (datasheet M05620; standard IHC practice). Check tissue preservation and the distribution of staining across the section before changing retrieval intensity, since uneven processing can complicate interpretation (standard IHC practice). Keep a consistently processed positive control in each run and report the fixation conditions alongside the staining result; a loss of signal alone cannot identify its cause.

Should RNF7 stain nuclei, cytoplasm, or both in IHC?

Assess nuclear and cytoplasmic staining separately: RNF7 is annotated in both compartments (UniProt Q9UBF6: cytoplasm and nucleus), while cell imaging places it mainly in the nucleoplasm with additional cytosolic localisation (HPA subcellular: supported). Tissue IHC reports cytoplasmic expression across several tissues, so cytoplasmic staining is also plausible in sections (HPA tissue IHC: cytoplasmic expression). Examine intact cells at high magnification with a counterstain and record which compartment contains chromogen, rather than scoring all colored area as equivalent (standard IHC practice). Diffuse staining that ignores cell boundaries needs comparison with the no-primary control and adjacent well-preserved tissue before assignment to RNF7.

Could RNF7 isoforms or epitope accessibility explain discordant staining?

RNF7 has 4 annotated isoforms, so determine which sequence the catalog antibody recognizes before comparing results across specimens (UniProt Q9UBF6: isoforms 1–4). Its annotated modifications include N-terminal acetylation at residue 2 and CK2 phosphorylation at residue 10; their effect on this antibody's binding is not established (UniProt Q9UBF6: modified residues). Ask for epitope information, then check whether that sequence is shared by the isoforms relevant to the experiment (standard IHC practice). If epitope information is unavailable, report the staining as antibody-detected RNF7 signal and avoid attributing differences to a particular isoform or modification.

How can IF help check the cell type and compartment behind an IHC signal?

Use IF as a separate validation experiment: pair RNF7 with a marker for the cell population identified on the IHC section, such as a tubular cell marker when assessing kidney tubules (HPA: High in kidney tubular cells; standard IF practice). Choose spectrally distinct fluorophores and inspect unstained tissue in each channel so autofluorescence does not masquerade as colocalisation (standard IF practice). RNF7 lacks a transmembrane segment and is reported in the nucleoplasm and cytosol, so permeabilise for intracellular epitope access; the antibody's precise epitope location is unspecified (UniProt Q9UBF6 topology; HPA subcellular). Compare compartment patterns with the chromogenic section, without transferring IHC retrieval or incubation settings directly to IF.

What should I change when RNF7 chromogen appears throughout the section?

Start with a no-primary control and inspect whether staining follows tissue edges, damaged areas or all cell types; those patterns warrant a background check before RNF7 scoring (standard IHC practice). The selected image used 3% BSA for 0.5 hour at room temperature and a 1:25 primary antibody dilution, providing documented starting conditions for this antibody (M05620 tissue-IHC caption). Check the peroxidase block and shorten chromogen development if the control also colors tissue (standard chromogenic IHC practice). Compare the result with the reported cytoplasmic tissue pattern and expected high staining in kidney tubular cells, while remembering that neither pattern alone proves antibody specificity (HPA tissue IHC).

How should I score RNF7 staining across samples? ⚠ ANSWER MARKED FOR VERIFICATION

Define the cell population and compartment before scoring; kidney tubular cells are a reported high-staining population, and tissue IHC describes predominantly cytoplasmic expression (HPA tissue IHC). For comparable regions, record the percentage of positive target cells and an H-score based on staining intensity, then report the denominator as all evaluable cells of that population (standard IHC scoring practice). If cells are sparse, positive-cell density per mm² can supplement the percentage; exclude folds, necrosis and section edges using the same rules in every sample (standard IHC practice). Keep retrieval, exposure to chromogen and scoring thresholds consistent, and present nuclear and cytoplasmic scores separately when both are observed.

When is an apparent RNF7-positive cell likely to be artefactual?

Treat a positive cell as more credible when staining respects cell boundaries and a plausible RNF7 compartment: UniProt lists nucleus and cytoplasm, and cell imaging supports nucleoplasm plus cytosol (UniProt Q9UBF6 localisation; HPA subcellular). Check cell identity rather than assuming every colored cell is expected; kidney tubular cells and bone marrow hematopoietic cells have reported high tissue staining (HPA tissue IHC). Signal confined to section edges, necrotic regions or the no-primary control needs investigation for processing or endogenous enzyme background (standard chromogenic IHC practice). HPA labels its tissue staining Approved but reports low consistency with RNA expression, so interpret isolated positives with matched controls and morphology (HPA tissue IHC reliability).

Boster reagents

Best RNF7 / RING-box protein 2 IHC Antibodies

RNF7 IHC has a human kidney paraffin-section image (M05620 image caption); human and mouse reactivity is listed (M05620 catalog). IF/ICC is listed for a second antibody without an image (M05620-1 catalog).

Real IHC data

Anti-RNF7 Antibody (N-term)

Cat # M05620

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M05620 is listed for IHC-P in human and mouse, with an IHC image from formaldehyde-fixed, paraffin-embedded human kidney sections (M05620 catalog; M05620 image caption). M05620-1 is listed for IHC and IF/ICC in human, mouse and rat, but has no IHC or IF image in the payload (M05620-1 catalog).

Which to pick: For paraffin-section tissue IHC, choose M05620 because its own image shows staining in formaldehyde-fixed human kidney sections (M05620 image caption); it is a rabbit polyclonal antibody (M05620 catalog). For IF/ICC, consider the rabbit monoclonal M05620-1, which lists those applications but provides no IF image; its IHC fixative is unreported (M05620-1 catalog). For rat samples, M05620-1 is the listed option; both SKUs list human and mouse reactivity (M05620-1 catalog; M05620 catalog).

Each figure is that product's own IHC / IF validation image from its datasheet.

References

UniProt Consortium. UniProt entry Q9UBF6 (RBX2_HUMAN, RING-box protein 2).
Human Protein Atlas. RNF7 tissue IHC expression (reliability: Approved).
Human Protein Atlas. RNF7 subcellular location (ICC-IF): Mainly localized to the nucleoplasm. In addition localized to the cytosol..
Human Protein Atlas. RNF7 antibody validation summary (1 antibodies).
RNF7 Facilitated the Tumorigenesis of Pancreatic Cancer by Activating PI3K/Akt Signaling Pathway. Oxidative medicine and cellular longevity 2023 — PMC9833898.
RNF7 inhibits apoptosis and sunitinib sensitivity and promotes glycolysis in renal cell carcinoma via the SOCS1/JAK/STAT3 feedback loop. Cellular & molecular biology letters 2022 — PMC9107170.
Cell type mapping of inflammatory muscle diseases highlights selective myofiber vulnerability in inclusion body myositis. Nature aging 2024 — PMC11257986.
RNF7-Mediated ROS Targets Malignant Phenotype and Radiotherapy Sensitivity in Glioma With Different IDH1 Genotypes. Molecular carcinogenesis 2025 — PMC11890431.
PubMed PMID:10512750 — UniProt-cited evidence.
PubMed PMID:10230407 — UniProt-cited evidence.
PubMed PMID:10082581 — UniProt-cited evidence.