This website uses cookies to ensure you get the best experience on our website.
- Table of Contents
NOB1 / RNA-binding protein NOB1 · IHC design guide
Design Immunohistochemistry for NOB1
Plan NOB1 staining in paraffin sections around the mainly nuclear tissue pattern (HPA tissue IHC). Use cell populations with documented medium or undetected staining as comparisons (HPA tissue IHC), and start with the catalog antibody at 2–5 μg/ml (datasheet A03504-1).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
NOB1 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 | Mainly nuclear in tissue sections (HPA tissue IHC) | |
| Staining pattern | Mainly nuclear staining across tissue cell types (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A03504-1) | |
| Positive control | Adrenal gland+4 more · see all | |
| Negative control | Adipose tissue+4 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A03504-1) | |
| Caveat | Some cell populations have no detectable staining (HPA tissue IHC) | |
| Regulation | No staining-linked regulator is reported (UniProt) | |
| Isoform / epitope | No reported isoforms; chain spans residues 2–412 (UniProt) |
Section 1
Recommended NOB1 IHC & IF Protocols
The catalog antibody’s IHC-P protocol is followed by 3 published NOB1 IHC protocols with usable preparation and staining details.
| Sample | Paraffin-embedded mouse brain tissue; fixative not specified (datasheet A03504-1) |
| Fixation | Image fixative and duration unreported (datasheet A03504-1); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat retrieval: EDTA pH 8.0 (datasheet A03504-1); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A03504-1) |
| Primary antibody | Rabbit anti-NOB1, 2-5 μg/ml (datasheet A03504-1) |
| Primary incubation | Overnight at 4 °C (datasheet A03504-1) |
| Detection | HRP-conjugated secondary, DAB chromogen (datasheet A03504-1) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | NOB1-positive staining in glandular cells of adrenal gland (HPA tissue IHC: Medium). HPA tissue profile: Mainly nuclear expression in all tissues. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA pH 8.0 retrieval (datasheet A03504-1); sodium citrate pH 6.0 is a published alternative (PMC4921844).
Section 2
What Is the Expected NOB1 Staining Pattern?
NOB1 is annotated as nuclear and has no transmembrane segment (UniProt Q9ULX3). In paraffin tissue IHC, expect mainly nuclear staining in the cell populations where HPA detects it, including glandular, respiratory epithelial, hematopoietic and neuronal cells (HPA tissue IHC). HPA rates the tissue staining Approved, with medium consistency between staining and RNA expression; interpret individual results with that limit in mind (HPA tissue IHC).
| Nuclear staining in adrenal or breast glandular cells, bronchial respiratory epithelial cells, bone marrow hematopoietic cells, or cortical neurons. | This matches HPA's mainly nuclear tissue pattern; each listed cell population has Medium staining (HPA tissue IHC). Judge staining in the identified cells, since HPA's observations are cell-specific rather than a promise that every cell in a section is positive (HPA tissue IHC). |
| Cytoplasmic-only or membrane-like staining dominates an otherwise interpretable paraffin section. | That differs from the mainly nuclear IHC pattern (HPA tissue IHC; UniProt Q9ULX3). Check morphology, counterstain and controls before calling it NOB1; HPA reports cytosol and focal adhesion sites in ICC-IF, a separate assay context (HPA subcellular ICC-IF). |
| Strong signal appears in cells listed as undetected, such as adipocytes or lung alveolar cells. | HPA reports NOB1 as Not detected in those specific cell types (HPA tissue IHC). An unexpected result warrants checking cell identification, nonspecific antibody binding and endogenous chromogen-related activity with controls (general IHC practice); it does not establish cross-reactivity by itself. |
| Brown signal spreads across tissue, empty spaces or many cell compartments without a clear nuclear pattern. | Treat this as background until controls distinguish specific staining from detection-system or antibody background (general IHC practice). HPA's mainly nuclear pattern provides a localisation reference, but its Approved rating has only medium consistency with RNA data (HPA tissue IHC). |
| No nuclear signal appears in a well-preserved section of an HPA Medium-positive cell population. | This is discordant with the reported IHC pattern for that population (HPA tissue IHC). Check the positive control and IHC workflow before assigning biological absence (general IHC practice); HPA's tissue observations do not establish how fixation or retrieval affects NOB1. |
💡Expected NOB1 appearanceCall a positive paraffin IHC result when identifiable HPA Medium-positive cells show mainly nuclear signal; dominant extranuclear staining or staining restricted to HPA-undetected cells needs control review (HPA tissue IHC; general IHC practice).
| IHC localisation and validation | HPA describes mainly nuclear expression across tissues and rates tissue IHC Approved, with medium staining-to-RNA consistency (HPA tissue IHC). Use the nuclear pattern as the primary slide reference while retaining that reliability caveat. |
| Choice of tissue and cell population | HPA reports Medium signal in several glandular, respiratory epithelial, hematopoietic and neuronal populations; adipocytes and lung alveolar cells are Not detected (HPA tissue IHC). Select controls by the reported cell population, rather than treating a whole organ as uniformly positive or negative. |
| Antibody-specific evidence | HPA049430 is IHC Approved; the other listed antibodies have no IHC status in this payload (HPA antibodies). HPA's tissue-level Approved rating does not turn an antibody without an IHC status into an IHC-validated reagent (HPA tissue IHC; HPA antibodies). |
| NOB1 topology and processing | UniProt places NOB1 in the nucleus, reports no transmembrane segment or signal peptide, and lists a chain spanning residues 2–412 (UniProt Q9ULX3). These annotations support intracellular interpretation; they do not identify this antibody's epitope or establish a retrieval condition. |
| What should IF/ICC show? | HPA's ICC-IF summary reports mainly cytosolic localisation, with focal adhesion sites additional; both locations are Approved (HPA subcellular ICC-IF). That differs from the mainly nuclear tissue IHC profile (HPA tissue IHC). HPA marks ICC Uncertain for HPA041522 and HPA067411 (HPA antibodies), so assess IF/ICC on its own guide page. |
| Retrieval and fixation evidence | No target-specific retrieval condition or fixation-sensitivity finding is supplied by UniProt or HPA here. Retrieval optimisation may be considered as general IHC practice, but HPA tissue staining levels cannot predict a fixation effect on NOB1. |
| Situation | Likely cause | Next action |
|---|---|---|
| No nuclear signal in a candidate positive section. | The chosen cells may be outside HPA's Medium-positive populations, or an IHC workflow step may have failed (HPA tissue IHC; general IHC practice). | Confirm the cell population and run a documented HPA Medium-positive population as a control (HPA tissue IHC); check detection reagents and the antibody's IHC instructions (general IHC practice). |
| Positive control stains, but the study sample is blank. | Expression may differ by cell population; HPA reports both Medium and Not detected populations (HPA tissue IHC). A blank sample alone cannot identify a fixation effect. | Score the relevant cells against their HPA cell-specific reference and confirm tissue morphology and control performance before calling the sample negative (HPA tissue IHC; general IHC practice). |
| Signal is mainly cytoplasmic in paraffin IHC. | That conflicts with HPA's mainly nuclear tissue profile, although cytosol is reported in ICC-IF (HPA tissue IHC; HPA subcellular ICC-IF). | Recheck cellular boundaries and counterstain, then compare with the IHC positive and reagent controls (general IHC practice); do not substitute the ICC-IF location for the IHC expectation. |
| Strong staining appears in adipocytes or alveolar cells. | Those cell types are Not detected in HPA tissue IHC; nonspecific binding or endogenous detection activity are possible explanations (HPA tissue IHC; general IHC practice). | Confirm cell identity and review appropriate negative detection controls alongside a nuclear positive control (general IHC practice; HPA tissue IHC). |
| The section has widespread diffuse brown background. | Incomplete blocking, excess antibody or endogenous chromogen-related activity can contribute to IHC background (general IHC practice); the appearance is unlike HPA's mainly nuclear pattern (HPA tissue IHC). | Review blocking, antibody dilution, washes and detection controls according to the assay instructions (general IHC practice). Do not score diffuse deposit as NOB1 localisation. |
| Results change after retrieval or fixation conditions change. | Processing differences can affect IHC performance in general (general IHC practice); no NOB1-specific fixation or retrieval effect is established by the supplied UniProt and HPA evidence. | Compare sections with the same positive control and documented processing conditions, and optimise the general IHC workflow if needed (general IHC practice). Avoid attributing the change specifically to NOB1 epitope masking. |
Experiment evidence
IHC & IF images showing NOB1 staining
A03504-1 · IHCIHC analysis of NOB1 using anti-NOB1 antibody (A03504-1). NOB1 was detected in a paraffin-embedded section of mouse brain tissue. Heat media
A03504-1 · IHCIHC analysis of NOB1 using anti-NOB1 antibody (A03504-1). NOB1 was detected in a paraffin-embedded section of rat brain tissue. Heat mediate
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000141101-NOB1/tissue/cerebral+cortex#imid_14589028
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000141101-NOB1/tissue/colon#imid_14600249
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000141101-NOB1/tissue/liver#imid_14600258
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000141101-NOB1/tissue/kidney#imid_14600267
HPA · ICC-IF A-549Human Protein Atlas immunofluorescence, A-549, antibody HPA041522; green = NOB1, blue = nucleus, red = microtubules.
HPA · ICC-IF A-549Human Protein Atlas immunofluorescence, A-549, antibody HPA041522; green = NOB1, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for NOB1 IHC & IF
🧪Run adrenal gland first and score its glandular cells for NOB1 staining (HPA: adrenal gland glandular cells, Medium). Use adipose tissue adipocytes as the negative comparison (HPA: adipose tissue adipocytes, Not detected); on the adrenal slide, compare staining with adjacent nonglandular cells as an internal background reference, without assuming those cells lack NOB1 (HPA: adrenal gland glandular cells, Medium).
Positive control tissue: Adrenal gland (Glandular cells, HPA Medium)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show NOB1 in A-549, Hep-G2, U2OS, CACO-2, with annotated localisation: Cytosol (approved) (HPA subcellular).
Technical controls: Include no-primary (secondary-only) and species-matched rabbit IgG controls, alongside NOB1-knockout material if available or peptide competition if the immunizing peptide is available (caption: rabbit anti-NOB1 antibody; standard IHC practice). Quench endogenous peroxidase and check background before interpreting DAB staining in the adrenal section (caption: peroxidase detection with DAB; standard chromogenic IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected SKU A03504-1 tissue-IHC caption does not state a fixative (caption: fixative not stated). The caption reports heat retrieval in EDTA at pH 8.0 for a paraffin mouse brain section; whether adrenal gland staining depends on that retrieval condition is unreported (caption: mouse brain IHC). HPA shows cytosolic and focal adhesion ICC-IF localization, while UniProt lists the nucleus, so assess localization carefully; the supplied evidence does not establish that frozen sections or IF are easier than paraffin IHC (HPA: subcellular ICC-IF; UniProt: nucleus).
HPA tissue IHC evidence for NOB1
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — Medium 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 |
|---|---|---|---|---|
| Adrenal gland | Glandular cells | Medium | Protein (IHC) | HPA → |
| Appendix | Glandular cells | Medium | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | Medium | Protein (IHC) | HPA → |
| Breast | Glandular cells | Medium | Protein (IHC) | HPA → |
| Bronchus | Respiratory epithelial cells | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Adipose tissue | Adipocytes | Not detected | Protein (IHC) | HPA → |
| Lung | Alveolar cells | Not detected | Protein (IHC) | HPA → |
| Lymph node | Germinal center cells | Not detected | Protein (IHC) | HPA → |
| Parathyroid gland | Glandular cells | Not detected | Protein (IHC) | HPA → |
| Prostate | Glandular cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced NOB1 IHC Tips
Troubleshoot NOB1 chromogenic IHC in paraffin sections by checking retrieval, staining compartment, controls, and how positive cells are scored.
Which retrieval conditions should I try first for weak NOB1 staining?
Start with heat-mediated retrieval in EDTA at pH 8.0 (datasheet A03504-1). That condition produced NOB1 staining in a paraffin-embedded mouse brain section, although the caption does not report the fixative (datasheet A03504-1). Keep section thickness, heating, cooling, and detection consistent while comparing retrieval runs, so changes in signal have an interpretable cause (standard IHC practice). If staining remains weak, test an alternative retrieval buffer only as a fallback, alongside the EDTA condition and a matched positive section (standard IHC practice). Excessive heating can damage morphology, so judge nuclear detail and background together with signal intensity (standard IHC practice).
Could fixation explain weak or uneven NOB1 IHC staining?
Target-specific fixation sensitivity for NOB1 is unknown from the supplied evidence: the selected paraffin-section caption does not state a fixative (datasheet A03504-1). Record the fixative, fixation duration, processing schedule, and section age before comparing staining across blocks (standard IHC practice). If samples differ, stain matched sections in one run using EDTA at pH 8.0 and the same detection settings (datasheet A03504-1; standard IHC practice). Inspect morphology and edge-to-center variation before changing antibody concentration, because processing differences can confound chromogenic comparisons (standard IHC practice). Do not attribute a fixation effect to NOB1 without a controlled comparison (standard IHC practice).
Should I score nuclear or cytoplasmic NOB1 staining in tissue?
Score nuclear and cytoplasmic signal separately: tissue IHC reports mainly nuclear expression, while subcellular IF reports approved cytosolic and focal-adhesion localization (HPA tissue IHC; HPA subcellular). UniProt also lists the nucleus, so neither compartment alone resolves the difference between assays (UniProt Q9ULX3). Review each pattern against cell morphology, a no-primary control, and staining in a documented positive tissue compartment (standard IHC practice; HPA tissue IHC). Report the fraction of cells staining in each compartment and keep the scoring rule fixed across specimens (standard IHC practice). Treat an isolated compartment shift as a finding requiring validation, rather than proof of NOB1 redistribution (standard IHC practice).
Could epitope choice or NOB1 processing explain discordant staining?
The record lists 0 annotated isoforms, a PINc domain at residues 5–108, and no transmembrane segment (UniProt Q9ULX3). It also records phosphorylation at residues 184, 201, 325, and 352, but supplies no antibody epitope map or evidence that these modifications alter staining (UniProt Q9ULX3; datasheet A03504-1). Ask for the immunogen or epitope information before assigning a staining discrepancy to processing or phosphorylation (standard IHC practice). Compare independently validated antibodies, if available, on adjacent sections with identical retrieval and detection (standard IHC practice). Record whether agreement concerns cell identity, staining compartment, and intensity, since agreement on intensity alone cannot establish epitope specificity (standard IHC practice).
How can I compare NOB1 IF with the chromogenic tissue result?
Use the IF result as a separate localization check: tissue IHC is mainly nuclear, whereas subcellular IF lists cytosol and focal adhesions (HPA tissue IHC; HPA subcellular). Multiplex NOB1 with a marker for the expected cell type, such as a neuronal-cell marker when assessing cerebral cortex neurons (HPA tissue IHC; standard IF practice). Choose spectrally separated fluorophores and consider a far-red channel if tissue autofluorescence obscures shorter wavelengths (standard IF practice). Apply permeabilisation appropriate for intracellular nuclear and cytosolic epitopes; NOB1 has no transmembrane segment, and its precise antibody epitope is unspecified (UniProt Q9ULX3; datasheet A03504-1). Check single-label and no-primary controls before comparing compartments across modalities (standard IF practice).
How do I distinguish diffuse DAB background from NOB1 signal?
The selected IHC caption used 10% goat serum blocking, 2 µg/mL primary antibody overnight at 4°C, and DAB development (datasheet A03504-1). Reproduce those conditions first, then inspect a no-primary section for secondary-reagent staining and tissue pigment (standard IHC practice). Include a peroxidase-blocking step and verify its effectiveness, because endogenous enzyme activity can produce chromogen independently of NOB1 (standard IHC practice). If diffuse staining persists, compare a primary-antibody titration while keeping retrieval, incubation, wash conditions, and DAB development constant (standard IHC practice). Count only cellular staining distinguishable from background and supported by the expected morphology (standard IHC practice; HPA tissue IHC).
What is a defensible way to quantify NOB1 IHC? ⚠ ANSWER MARKED FOR VERIFICATION
Define the analysis region and score nuclear and cytoplasmic staining separately, because tissue IHC and subcellular IF report different predominant compartments (HPA tissue IHC; HPA subcellular). For cellular staining, record the percentage of positive cells and, if intensity is reliable, an H-score from 0–300 using fixed intensity bins (standard IHC practice). Normalize cell counts to the number of eligible cells; use positive cells per mm² only when area-based density answers the biological question (standard IHC practice). Apply one threshold, counterstain standard, and exclusion rule across sections (standard IHC practice). Report cell type and compartment alongside the score so mixed populations do not conceal a localized change (standard IHC practice).
When should an apparent NOB1-positive area be treated as artefact?
First ask whether staining follows identifiable cells and whether its compartment is plausible: tissue IHC is mainly nuclear, while IF also supports cytosolic and focal-adhesion localization (HPA tissue IHC; HPA subcellular). Compare candidate positive cells with documented medium-staining neuronal cells in cerebral cortex or cerebellar molecular-layer cells, while recognizing that these references do not validate every specimen (HPA tissue IHC). Discount staining concentrated at section edges, in necrotic areas, or in no-primary controls (standard IHC practice). Check peroxidase-blocked controls when DAB signal could reflect endogenous enzyme activity (standard IHC practice). Confirm an unexpected cell type or compartment with an independent specificity control before interpreting it biologically (standard IHC practice).
Boster reagents
Best NOB1 / RNA-binding protein NOB1 IHC Antibodies
A03504-1 has IHC images from paraffin-embedded mouse and rat brain and human liver sections (catalog image captions). No IF/ICC data are supplied (catalog applications and images).
Real IHC data
Anti-NOB1 Antibody ®
Cat # A03504-1
A03504-1 is the only listed antibody; its IHC images show paraffin-embedded mouse and rat brain and human liver sections (catalog image captions). Its listed applications include IHC, while IF/ICC is absent (catalog applications).
Which to pick: Choose A03504-1 for paraffin-section IHC: its own captions document those three tissues, and the fixative is unreported (catalog image captions). No listed SKU has IF/ICC validation, so there is no supported IF/ICC pick (catalog applications and images). A03504-1 lists human, mouse and rat reactivity and a rabbit host; clonality is unreported (catalog reactivity, host and clone fields).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q9ULX3 (NOB1_HUMAN, RNA-binding protein NOB1).
- Human Protein Atlas. NOB1 tissue IHC expression (reliability: Approved).
- Human Protein Atlas. NOB1 subcellular location (ICC-IF): Mainly localized to the cytosol. In addition localized to the focal adhesion sites..
- Human Protein Atlas. NOB1 antibody validation summary (3 antibodies).
- Downregulated miR-646 in clear cell renal carcinoma correlated with tumour metastasis by targeting the nin one binding protein (NOB1). British journal of cancer 2014 — PMC4453839.
- Long non-coding RNA SNHG1 regulates NOB1 expression by sponging miR-326 and promotes tumorigenesis in osteosarcoma. International journal of oncology 2018 — PMC5743365.
- High Expression of RIOK2 and NOB1 Predict Human Non-small Cell Lung Cancer Outcomes. Scientific reports 2016 — PMC4921844.
- MicroRNA-326 functions as a tumor suppressor in glioma by targeting the Nin one binding protein (NOB1). PloS one 2013 — PMC3711818.
- PubMed PMID:16172919 — UniProt-cited evidence.
- PubMed PMID:14702039 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for NOB1 (RNA-binding protein NOB1). Expected localisation Mainly nuclear in tissue sections (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A03504-1). Validated IHC antibody: A03504-1 at 2-5 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Adrenal gland (Glandular cells); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded mouse brain tissue; fixative not specified (datasheet A03504-1); 2. Fixation: Image fixative and duration unreported (datasheet A03504-1); verify before use.; 3. Sectioning: 4–5 µm sections on charged slides (standard); 4. Deparaffinisation: Xylene, graded ethanol series to water (standard); 5. Antigen retrieval: Heat retrieval: EDTA pH 8.0 (datasheet A03504-1); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A03504-1); 8. Primary antibody: Rabbit anti-NOB1, 2-5 μg/ml (datasheet A03504-1); 9. Primary incubation: Overnight at 4 °C (datasheet A03504-1). Watch for: Fixation: Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A03504-1); Caveat: Some cell populations have no detectable staining (HPA tissue IHC); Isoform / epitope: No reported isoforms; chain spans residues 2–412 (UniProt); Regulation: No staining-linked regulator is reported (UniProt). Evidence: UniProt Q9ULX3, HPA tissue IHC · Approved, HPA ICC-IF · Mainly localized to the cytosol. In addition localized to the focal…, Datasheet A03504-1, PMC4453839, PMC4921844. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/NOB1/immunohistochemistry-ihc-guide.