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NIBAN2 / Protein Niban 2 · IHC design guide
Design Immunohistochemistry for NIBAN2
Plan NIBAN2 staining in paraffin sections with the catalog antibody at 2–5 μg/ml (datasheet A09391-2). Assess cytoplasmic and membranous staining using colon or stomach glandular cells as positive references, while accounting for uncertain tissue staining reliability (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
NIBAN2 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 and membranous tissue staining (HPA tissue IHC) | |
| Staining pattern | Most tissues: cytoplasmic and membranous staining (HPA tissue IHC) | |
| Antigen retrieval | EDTA pH 8.0 HIER, heat-mediated (datasheet A09391-2) | |
| Positive control | Appendix+4 more · see all | |
| Negative control | Bone marrow+4 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep paraffin-section fixation consistent across samples. (standard IHC practice; not target-specific) | |
| Caveat | Tissue staining reliability is uncertain (HPA tissue IHC) | |
| Regulation | Expression regulation is not annotated (UniProt) | |
| Isoform / epitope | 2 isoforms; check epitope coverage (UniProt) |
Section 1
Recommended NIBAN2 IHC & IF Protocols
The catalog antibody protocol is paired with one published NIBAN2 tissue staining protocol (PMC12432436).
| Sample | Paraffin-embedded human placenta tissue; fixative not specified (datasheet A09391-2) |
| Fixation | Image fixative and duration unreported (datasheet A09391-2); 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 A09391-2); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A09391-2) |
| Primary antibody | Rabbit anti-NIBAN2, 2-5 μg/ml (datasheet A09391-2) |
| Primary incubation | Overnight at 4 °C (datasheet A09391-2) |
| Detection | Streptavidin-biotin complex (SABC), DAB chromogen (datasheet A09391-2) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | NIBAN2-positive staining in glandular cells of appendix (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic and membranous expression in most tissues. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated EDTA retrieval at pH 8.0 (datasheet A09391-2); the published protocol does not specify retrieval (PMC12432436).
Section 2
What Is the Expected NIBAN2 Staining Pattern?
NIBAN2 staining is expected in the cytoplasm and at cell membranes, including adherens junctions (UniProt Q96TA1; HPA tissue IHC). Strong examples include glandular cells of the colon, stomach, appendix, duodenum, endometrium and gallbladder, and urothelial cells of the urinary bladder (HPA: High). NIBAN2 has no transmembrane segment (UniProt Q96TA1 topology). Treat the tissue pattern as provisional: HPA rates its IHC reliability Uncertain, with medium agreement between staining and RNA expression (HPA tissue IHC).
| Cytoplasmic staining, with some membrane accentuation, in glandular or urothelial cells (HPA tissue IHC). | This fits the reported pattern; colon glandular cells and urinary bladder urothelial cells are examples rated High (HPA tissue IHC). Compare the distribution within the tissue, since HPA reports staining in most tissues but rates the IHC evidence Uncertain (HPA tissue IHC). |
| Predominantly nuclear staining in a paraffin tissue section. | This differs from the principal cytoplasmic and membranous tissue pattern (HPA tissue IHC). Review morphology, counterstain and controls for an artefact (general IHC practice). HPA reports additional nucleoplasmic signal in ICC-IF, so nuclear signal alone is not definitive proof of antibody cross-reactivity (HPA subcellular ICC-IF). |
| Strong signal mainly in cells recorded as Not detected, such as skeletal muscle myocytes (HPA tissue IHC). | This conflicts with that cell-type reference and merits a control check; cross-reactivity or endogenous detection activity are possibilities, not established diagnoses (HPA tissue IHC; general IHC practice). A Not detected entry applies to the named cells, not necessarily every cell in that tissue (HPA tissue IHC). |
| Uniform haze over cells and surrounding tissue, without a discernible cellular pattern. | This is less interpretable than compartment-resolved staining (HPA tissue IHC; general IHC practice). Examine a no-primary control and staining run for nonspecific background or endogenous detection activity before assigning NIBAN2 positivity (general IHC practice). |
| No signal in colon glandular cells included as a positive reference (HPA: High in colon glandular cells). | Check section integrity, retrieval, detection and the run controls before calling the specimen negative (general IHC practice). The HPA High rating makes these cells a useful reference, but HPA's overall IHC reliability is Uncertain (HPA tissue IHC). |
💡Expected NIBAN2 appearanceA convincing positive shows High cytoplasmic staining with possible membrane accentuation in the listed glandular or urothelial cells; isolated diffuse haze or strong signal in an HPA Not detected cell type warrants investigation (HPA tissue IHC; general IHC practice).
| Cell state and compartment (UniProt Q96TA1). | UniProt reports exclusively cytoplasmic localization in exponentially growing cells and membrane localization at confluency or during telophase; a tissue section need not show identical membrane emphasis in every cell (UniProt Q96TA1). |
| Tissue and cell-type reference (HPA tissue IHC). | HPA rates selected glandular and urothelial cells High, placental cytotrophoblasts Medium, and selected other cell types Not detected. Interpret the named cell type rather than treating an entire organ as uniformly positive or negative (HPA tissue IHC). |
| IHC evidence strength (HPA tissue IHC; HPA antibodies). | HPA calls the tissue IHC profile Uncertain, pending external verification; all four listed HPA antibodies have IHC status Uncertain. Use the pattern as an expectation, not a stand-alone specificity test (HPA tissue IHC; HPA antibodies). |
| Topology and processing (UniProt Q96TA1). | NIBAN2 has no transmembrane segment, signal peptide or annotated propeptide; these annotations provide no basis for predicting a shed extracellular staining pattern or a cleavage-dependent IHC result (UniProt Q96TA1 topology and processing). |
| Isoforms and phosphorylation (UniProt Q96TA1). | UniProt lists two isoforms and multiple modified residues. Antibody epitope and isoform coverage are not supplied here, so neither feature establishes which staining differences this IHC antibody can detect (UniProt Q96TA1). |
| IF/ICC: what localization is reported? (HPA subcellular ICC-IF). | HPA reports enhanced plasma membrane and cytosol localization, plus supported nucleoplasmic localization, in ICC-IF; that assay's compartment evidence should not be treated as an IHC validation result (HPA subcellular ICC-IF; HPA antibodies). |
| Situation | Likely cause | Next action |
|---|---|---|
| Known-positive glandular cells show no chromogenic signal (HPA: High in colon glandular cells). | Target-specific fixation effects are not established by the supplied assay evidence. Verify with a matched IHC source before attributing a result to fixation. | Confirm tissue morphology and run controls, then review the validated IHC workflow, including retrieval and detection, before interpreting a negative result (general IHC practice). |
| Strong nuclear-only staining dominates the section. | It departs from the principal tissue pattern, although ICC-IF reports additional nucleoplasmic localization (HPA tissue IHC; HPA subcellular ICC-IF). | Check the no-primary control, nuclear counterstain and cellular boundaries; require matching cytoplasmic or membranous tissue context before a confident IHC call (general IHC practice; HPA tissue IHC). |
| The whole section has brown haze that obscures cell borders. | Nonspecific background or endogenous detection activity may obscure compartment assignment (general IHC practice). | Review no-primary and detection controls, blocking and reagent handling, then assess whether cell-resolved cytoplasmic or membrane staining becomes visible (general IHC practice; HPA tissue IHC). |
| A nominally negative tissue contains a small stained population. | HPA Not detected entries refer to specified cells, such as bone marrow hematopoietic cells, and do not classify every cell in the organ (HPA tissue IHC). | Identify the stained cell population morphologically and compare that cell type with HPA's entry before labeling the tissue result discordant (HPA tissue IHC; general IHC practice). |
| Staining is strongest in an HPA Not detected cell type (HPA tissue IHC). | Cross-reactivity or endogenous detection activity is possible; HPA's IHC reliability is Uncertain, so the reference alone cannot resolve the cause (HPA tissue IHC; general IHC practice). | Compare positive-reference tissue and no-primary controls from the same run, and seek independent specificity evidence before accepting that signal as NIBAN2 (general IHC practice). |
| Some positive cells show little membrane accentuation. | UniProt describes cytoplasmic localization in growing cells and membrane localization with confluency or telophase; HPA describes both cytoplasmic and membranous tissue expression (UniProt Q96TA1; HPA tissue IHC). | Score cytoplasmic and membrane compartments separately, using the named positive cell types as context; do not require a membrane rim in every cell (HPA tissue IHC; general IHC practice). |
Experiment evidence
IHC & IF images showing NIBAN2 staining
A09391-2 · IHCIHC analysis of FAM129B/NIBAN2 using anti-FAM129B/NIBAN2 antibody (A09391-2). FAM129B/NIBAN2 was detected in a paraffin-embedded section of
A09391-2 · IHCIHC analysis of FAM129B/NIBAN2 using anti-FAM129B/NIBAN2 antibody (A09391-2). FAM129B/NIBAN2 was detected in a paraffin-embedded section of
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000136830-NIBAN2/tissue/cerebral+cortex#imid_6359272
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000136830-NIBAN2/tissue/colon#imid_6360134
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000136830-NIBAN2/tissue/liver#imid_6360135
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000136830-NIBAN2/tissue/kidney#imid_6360128
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA021284; green = NIBAN2, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA021284; green = NIBAN2, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for NIBAN2 IHC & IF
🧪Run colon first; its glandular cells should stain (HPA: High in colon glandular cells). Use bone marrow hematopoietic cells as the negative tissue (HPA: Not detected in bone marrow hematopoietic cells); on the colon slide, assess background in nonglandular areas, but do not assume those cells are NIBAN2-negative from the supplied HPA row.
Positive control tissue: Appendix (Glandular cells, HPA High)
Negative control tissue: Bone marrow (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show NIBAN2 in A-431, U-251MG, U2OS, NIH 3T3, with annotated localisation: Plasma membrane (enhanced), Cytosol (enhanced) (HPA subcellular).
Technical controls: Include a no-primary (secondary-only) slide, an isotype control matched to the rabbit primary antibody, and a NIBAN2 knockout sample as a biological negative (caption: rabbit anti-NIBAN2 antibody; standard IHC practice). For colon sections developed with SABC/DAB, control endogenous peroxidase and check endogenous biotin background (caption: SABC/DAB detection; standard IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected A09391-2 tissue-IHC caption does not report a fixative (selected-SKU caption: fixative not stated). The caption uses heat retrieval in EDTA at pH 8.0, but does not establish that retrieval is required (selected-SKU caption: heat-mediated EDTA retrieval). IF/ICC images exist for this protein, but the supplied evidence does not show that IF or frozen sections are easier than paraffin IHC; in colon, check endogenous peroxidase and biotin background with the caption’s SABC/DAB method (HPA: ICC-IF images; caption: SABC/DAB detection; standard IHC practice).
HPA tissue IHC evidence for NIBAN2
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Uncertain — Medium consistency between antibody staining and RNA expression data. Pending external verification.). 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 |
|---|---|---|---|---|
| Appendix | Glandular cells | High | Protein (IHC) | HPA → |
| Colon | Glandular cells | High | Protein (IHC) | HPA → |
| Duodenum | Glandular cells | High | Protein (IHC) | HPA → |
| Endometrium | Glandular cells | High | Protein (IHC) | HPA → |
| Gallbladder | Glandular cells | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Bone marrow | Hematopoietic cells | Not detected | Protein (IHC) | HPA → |
| Caudate | Glial cells | Not detected | Protein (IHC) | HPA → |
| Cerebellum | Cells in granular layer | Not detected | Protein (IHC) | HPA → |
| Hippocampus | Glial cells | Not detected | Protein (IHC) | HPA → |
| Lymph node | Germinal center cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced NIBAN2 IHC Tips
Use the documented paraffin-section workflow as the starting point, then assess NIBAN2 staining by cell type and compartment (caption A09391-2; HPA tissue IHC).
How should I optimize retrieval when NIBAN2 staining is weak in paraffin sections?
Start with heat-mediated antigen retrieval in EDTA at pH 8.0 for paraffin sections (datasheet A09391-2). The selected placenta image used this condition with 10% goat serum blocking and 2 μg/ml primary overnight at 4°C (caption A09391-2). Keep heating and cooling times consistent, and compare a positive section with a no-primary control while adjusting retrieval duration (standard IHC practice). Score cytoplasm and cell borders separately, matching the reported NIBAN2 compartments rather than accepting diffuse nuclear color (UniProt Q96TA1 localisation; HPA tissue IHC). If weak, adjust heating time before testing another buffer, documenting morphology and background (standard IHC practice).
Could fixation explain variable NIBAN2 staining between paraffin blocks?
Target-specific sensitivity to fixation type or duration is unknown because the selected NIBAN2 caption does not state a fixative (caption A09391-2: fixative not stated). Record fixative, exposure time, cold ischemia, section age, and processing history for each block before comparing staining intensity (standard IHC practice). Use similarly processed control blocks, and inspect preservation of cellular borders and nuclei before interpreting a weak result (standard IHC practice). Apply the documented EDTA pH 8.0 retrieval consistently while changing other variables one at a time (datasheet A09391-2; standard IHC practice). Neither tissue staining patterns nor phosphorylation annotations establish fixation sensitivity (HPA tissue IHC; UniProt Q96TA1 modified residues).
Should NIBAN2 appear in cytoplasm, at cell borders, or in nuclei?
Expect cytoplasmic and membranous staining in many tissues (HPA tissue IHC), consistent with cytosol and adherens junction annotation (UniProt Q96TA1 localisation). Distinguish cell-border signal from cytoplasmic DAB by examining intact adjacent cells at the same development time and with matched controls (standard IHC practice). NIBAN2 can be exclusively cytoplasmic during exponential growth and shift toward the membrane at confluency or telophase; use that observation to guide interpretation without assigning a fixed membrane fraction to tissue sections (UniProt Q96TA1 localisation). Treat isolated nuclear staining cautiously: additional nucleoplasmic localisation is reported, while the tissue IHC reliability rating remains uncertain (HPA subcellular; HPA tissue IHC reliability).
Can this stain distinguish NIBAN2 isoforms or phosphorylation states?
NIBAN2 has 2 annotated isoforms and a PH domain at residues 68–192 (UniProt Q96TA1 isoforms; domains). Because the catalog antibody's epitope is not mapped in the supplied evidence, do not assume equal recognition of both isoforms or a phosphorylation-independent readout (provided antibody evidence). Modified residues include sites near 568–606, so recognition of an epitope in that region would need separate testing (UniProt Q96TA1 modified residues; standard IHC validation practice). No transmembrane segment is annotated despite membrane localisation; cell-border staining therefore does not establish that the antibody recognizes an extracellular epitope (UniProt Q96TA1 topology; localisation).
How should I assess NIBAN2 by multiplex IF alongside this IHC workflow?
For a separate IF/ICC experiment, pair NIBAN2 with a validated marker of the expected cell type, such as a cytotrophoblast marker in placenta (HPA tissue IHC: medium in placental cytotrophoblasts; standard IF practice). Select a far-red fluorophore and include unstained and single-color controls to assess tissue autofluorescence and channel bleed-through (standard IF practice). Compare permeabilized and nonpermeabilized preparations because the catalog antibody's epitope accessibility is unspecified and NIBAN2 lacks a transmembrane segment (provided antibody evidence; UniProt Q96TA1 topology). Titrate permeabilisation when seeking cytosolic signal, and evaluate cell-border staining separately because both compartments are reported (UniProt Q96TA1 localisation; standard IF practice).
How can I separate NIBAN2 staining from chromogenic background?
Start with a no-primary section and a secondary-only control to locate nonspecific DAB deposition (standard IHC practice). The selected procedure uses biotinylated goat anti-rabbit IgG, an avidin-biotin complex, and DAB, so assess endogenous biotin and peroxidase contributions (caption A09391-2; standard IHC practice). Block endogenous peroxidase and consider an avidin-biotin blocking step if control sections retain color (standard IHC practice). Retitrate the primary around the documented 2 μg/ml and verify that the 10% goat serum block is applied uniformly (caption A09391-2; standard IHC practice). Shorten DAB development when diffuse color masks cytoplasm or cell borders (standard IHC practice).
How should I quantify NIBAN2 staining across paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION
Score NIBAN2 within prespecified cell types and compartments, because cytoplasmic and membranous expression is reported in most tissues (HPA tissue IHC). For chromogenic sections, record percent positive cells and intensity, then calculate an H-score on a 0–300 scale from percentages assigned 0–3 intensity (standard IHC scoring practice). Alternatively, report positive cell density per mm² of viable tissue, using the same segmentation and DAB threshold across batches (standard image-analysis practice). Normalize counts to evaluable target cells or viable tissue area, and report cytoplasmic and membrane scores separately (standard IHC practice; UniProt Q96TA1 localisation).
What distinguishes a credible NIBAN2-positive section from an artefact?
Look for staining in viable cytoplasm or along cell borders, consistent with NIBAN2 localisation, and confirm it with a processed control section (UniProt Q96TA1 localisation; standard IHC practice). Check cell identity: glandular cells in appendix and colon show high staining, whereas bone-marrow hematopoietic cells were not detected in the supplied atlas (HPA tissue IHC). Treat staining restricted to edges, folds, necrotic areas, or nuclei as uncertain until repeat sections establish its distribution; additional nucleoplasmic localisation has been reported (standard IHC practice; HPA subcellular). Use no-primary and peroxidase-block controls to investigate endogenous enzyme or detection background in the documented DAB workflow (caption A09391-2; standard IHC practice).
Boster reagents
Best NIBAN2 / Protein Niban 2 IHC Antibodies
The catalog antibody has real IHC data from human paraffin sections (A09391-2 image captions); its listed reactivity also includes mouse and rat (A09391-2 catalog). No IF data are supplied (A09391-2 catalog).
Real IHC data
Anti-FAM129B/NIBAN2 Antibody ®
Cat # A09391-2
A09391-2 was tested by IHC on paraffin sections of human placenta and human lung cancer tissue (A09391-2 image captions). Its listed applications include IHC, while no IF image or IF dilution is supplied (A09391-2 catalog).
Which to pick: Choose A09391-2 for human paraffin-section IHC: its captions show EDTA retrieval at pH 8.0 and primary antibody at 2 μg/ml (A09391-2 image captions); the fixative is unreported (A09391-2 image captions). No SKU in the payload is documented for IF/ICC (A09391-2 catalog). A09391-2 lists mouse and rat reactivity, but the supplied IHC images show human tissue only (A09391-2 catalog; A09391-2 image captions).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q96TA1 (NIBA2_HUMAN, Protein Niban 2).
- Human Protein Atlas. NIBAN2 tissue IHC expression (reliability: Uncertain).
- Human Protein Atlas. NIBAN2 subcellular location (ICC-IF): Mainly localized to the plasma membrane and cytosol. In addition localized to the nucleoplasm..
- Human Protein Atlas. NIBAN2 antibody validation summary (4 antibodies).
- NIBAN2 Stimulates Glioma Growth by Regulating the JAK2/STAT3/c-Myc Pathway. Cancer medicine 2025 — PMC12432436.
- NIBAN2/FLII/RREB1 Axis Drives Glioma Stem Cell Malignancy via TLR3 Pathway Activation. Advanced science (Weinheim, Baden-Wurttemberg, Germany) 2026 — PMC13159147.
- PubMed PMID:15164053 — UniProt-cited evidence.
- PubMed PMID:17974005 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for NIBAN2 (Protein Niban 2). Expected localisation Cytoplasmic and membranous tissue staining (HPA tissue IHC). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A09391-2). Validated IHC antibody: A09391-2 at 2-5 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Appendix (Glandular cells); negative Bone marrow. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human placenta tissue; fixative not specified (datasheet A09391-2); 2. Fixation: Image fixative and duration unreported (datasheet A09391-2); 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 A09391-2); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A09391-2); 8. Primary antibody: Rabbit anti-NIBAN2, 2-5 μg/ml (datasheet A09391-2); 9. Primary incubation: Overnight at 4 °C (datasheet A09391-2). Watch for: Fixation: Keep paraffin-section fixation consistent across samples. (standard IHC practice; not target-specific); Caveat: Tissue staining reliability is uncertain (HPA tissue IHC); Isoform / epitope: 2 isoforms; check epitope coverage (UniProt); Regulation: Expression regulation is not annotated (UniProt). Evidence: UniProt Q96TA1, HPA tissue IHC · Uncertain, HPA ICC-IF · Mainly localized to the plasma membrane and cytosol. In addition…, Datasheet A09391-2, PMC12432436. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/NIBAN2/immunohistochemistry-ihc-guide.