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SREBF2 / Sterol regulatory element-binding protein 2 · IHC design guide
Design Immunohistochemistry for SREBF2
Plan paraffin-section SREBF2 IHC around the nuclear staining reported in most tissues (HPA tissue IHC). Colon glandular cells show high staining (HPA tissue IHC), while sterol-dependent processing can affect nuclear signal (UniProt).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
SREBF2 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 | Nuclear staining in most tissues (HPA tissue IHC); ER/Golgi precursor (UniProt) | |
| Staining pattern | Nuclear staining across most tissue cell types (HPA tissue IHC) | |
| Antigen retrieval | Tris-EDTA pH 9.0 HIER, 95–98 °C, 20 min (rule: nuclear antigen) | |
| Positive control | Appendix+4 more · see all | |
| Negative control | Cerebellum+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 A01678) | |
| Caveat | Sterol status can change processing and nuclear signal (UniProt) | |
| Regulation | Low sterols promote nuclear SREBF2 (UniProt) | |
| Isoform / epitope | 2 isoforms; processed 1–484 lacks C-terminal epitopes (UniProt) |
Section 1
Recommended SREBF2 IHC & IF Protocols
The catalog antibody’s IHC-P protocol is accompanied by published paraffin-section methods for colon adenocarcinoma and cervical tumors (PMC10723182; PMC12992596).
| Sample | Tissue sections; selected-image fixative not specified (standard IHC workflow) |
| Fixation | Image fixative and duration unreported (datasheet A01678); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat-induced epitope retrieval in Tris-EDTA buffer, pH 9.0, 20 min at 95–98 °C (standard rule: nuclear antigen) |
| 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-SREBF2, 5 μg/mL (datasheet A01678) |
| Primary incubation | Overnight at 4 °C (standard) |
| Detection | HRP-polymer secondary, DAB chromogen 5–10 min (standard) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | SREBF2-positive staining in lymphoid tissue of appendix (HPA tissue IHC: High). HPA tissue profile: Nuclear expression in most tissues. No signal in the no-primary control. |
💡Decision noteStart with Tris-EDTA pH 9.0 at 95–98 °C for 20 min (page retrieval rule: nuclear antigen); compare the published 10 min retrieval if needed (PMC12992596).
Section 2
What Is the Expected SREBF2 Staining Pattern?
SREBF2 occupies ER and Golgi membranes and can enter nuclei after processing (UniProt Q12772: localization, processing). HPA reports nuclear staining in most tissues, including high staining in colon glandular cells (HPA: tissue IHC). Its IHC profile is Approved, with low consistency between antibody staining and RNA expression (HPA: reliability).
| High nuclear stain in colon glands (HPA: tissue IHC). | Expected pattern; HPA reports nuclear expression in most tissues (HPA: tissue IHC). |
| Stain confined to extracellular spaces (general IHC practice). | Suspect artefact; SREBF2 localizes inside cells (UniProt Q12772: localization). |
| Strong stain in pancreatic exocrine cells (HPA: not detected). | Check cross-reactivity or endogenous detection activity (general IHC practice). |
| Diffuse stain across tissue and blank areas (general IHC practice). | Background is likely; compare a no-primary control (general IHC practice). |
| No stain in colon glands (HPA: High). | Review the assay before scoring these cells negative (general IHC practice). |
💡Expected SREBF2 appearanceCall colon glands positive when nuclear staining is high (HPA: tissue IHC); blank-area DAB suggests background (general IHC practice).
| Sterol-dependent processing (UniProt Q12772). | Low sterol releases a nuclear form; high sterol retains the precursor in ER (UniProt: processing). |
| Membrane topology (UniProt Q12772). | The precursor has 2 transmembrane spans; the processed chain is residues 1–484 (UniProt: topology). |
| IHC evidence (HPA: antibodies). | HPA031962 is IHC Approved; tissue staining has low RNA consistency (HPA: antibodies, reliability). |
| IF/ICC: where is signal? | Nucleoplasm and vesicles; ER also seen (HPA: supported ICC-IF localization). |
| Situation | Likely cause | Next action |
|---|---|---|
| No nuclear DAB in colon glands (HPA: High). | Assay failure is possible (general IHC practice). | Check positive control, retrieval and detection steps (general IHC practice). |
| Signal in a no-primary section (general IHC practice). | Endogenous activity or reagent background (general IHC practice). | Block endogenous activity and repeat the control (general IHC practice). |
| Diffuse tissue haze (general IHC practice). | Blocking or washing may be inadequate (general IHC practice). | Adjust blocking, antibody dilution and washes (general IHC practice). |
| Strong prostate gland stain (HPA: not detected). | Cross-reactivity or detection background is possible (general IHC practice). | Compare no-primary control and verify antibody specificity (general IHC practice). |
| Cytoplasmic stain without nuclear stain (HPA: nuclear in most tissues). | ER precursor may contribute (UniProt Q12772: localization, processing). | Check counterstain and score compartments separately (general IHC practice). |
Experiment evidence
IHC & IF images showing SREBF2 staining
A01678 · IHCImmunohistochemistry of SREBF2 in human spleen tissue with SREBF2 antibody at 5 μg/mL.
A01678 · IFImmunofluorescence of SREBF2 in human spleen tissue with SREBF2 antibody at 20 μg/mL.
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000198911-SREBF2/tissue/cerebral+cortex#imid_8026455
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000198911-SREBF2/tissue/colon#imid_8026373
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000198911-SREBF2/tissue/liver#imid_8026374
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000198911-SREBF2/tissue/kidney#imid_8026383
HPA · ICC-IF HEK293Human Protein Atlas immunofluorescence, HEK293, antibody HPA031963; green = SREBF2, blue = nucleus, red = microtubules.
HPA · ICC-IF HEK293Human Protein Atlas immunofluorescence, HEK293, antibody HPA031963; green = SREBF2, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for SREBF2 IHC & IF
🧪Run colon first: glandular cells should stain (HPA: High in colon glandular cells). Use heart muscle as the negative tissue; cardiomyocytes are not detected (HPA: Not detected in heart muscle cardiomyocytes). On the colon slide, morphologically unstained neighboring cells should remain at background, but their SREBF2 status is unreported.
Positive control tissue: Appendix (Lymphoid tissue, HPA High)
Negative control tissue: Cerebellum (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show SREBF2 in HEK293, SK-MEL-30, U2OS, with annotated localisation: Nucleoplasm (supported), Vesicles (supported) (HPA subcellular).
Technical controls: Include no-primary (secondary-only) and host-species- and clonality-matched isotype controls, plus a SREBF2 knockout specimen or an immunizing-peptide block if available (standard IHC practice). Quench endogenous peroxidase in colon before chromogenic detection; assess endogenous biotin if using avidin–biotin detection (standard IHC practice).
⚠️Feasibility: A target-specific fixation window and fixation effect are unreported, and the selected A01678 spleen IHC caption does not state a fixative (selected-SKU caption). Retrieval dependency is unreported; optimize antigen retrieval on paraffin sections (standard IHC practice). The supplied evidence does not establish whether frozen sections or IF are easier; SREBF2 can occupy nuclear, vesicular and ER compartments, so score compartment-specific staining rather than treating every pattern as equivalent (HPA: supported nucleoplasm, vesicles and ER; UniProt Q12772 processing).
HPA tissue IHC evidence for SREBF2
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 |
|---|---|---|---|---|
| Appendix | Lymphoid tissue | High | Protein (IHC) | HPA → |
| Bone marrow | Hematopoietic cells | High | Protein (IHC) | HPA → |
| Colon | Glandular cells | High | Protein (IHC) | HPA → |
| Duodenum | Glandular cells | High | Protein (IHC) | HPA → |
| Esophagus | Squamous epithelial cells | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Cerebellum | Cells in granular layer | Not detected | Protein (IHC) | HPA → |
| Heart muscle | Cardiomyocytes | Not detected | Protein (IHC) | HPA → |
| Liver | Cholangiocytes | Not detected | Protein (IHC) | HPA → |
| Pancreas | Exocrine glandular cells | Not detected | Protein (IHC) | HPA → |
| Parathyroid gland | Glandular cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced SREBF2 IHC Tips
Troubleshoot chromogenic SREBF2 IHC by checking retrieval, compartment-specific staining and cell-level controls before interpreting signal as sterol-dependent processing.
How should I retrieve SREBF2 in paraffin sections with weak nuclear staining?
Use Tris-EDTA pH 9.0 heat-induced epitope retrieval at 95–98 °C for 20 min (page retrieval setting). Let sections cool in the retrieval solution, then compare a matched positive tissue section with a no-primary control under the same staining conditions (standard IHC practice). If staining remains weak, test a shorter or longer heating time on adjacent sections while keeping antibody concentration and detection constant; excessive heating can damage morphology (standard IHC practice). Score nuclear and cytoplasmic signal separately because SREBF2 has a processed nuclear form and a membrane-associated precursor (UniProt Q12772 processing and subcellular location).
Could fixation explain absent SREBF2 staining in my paraffin sections?
Target-specific fixation sensitivity is unknown: the selected SREBF2 spleen IHC caption reports 5 µg/mL antibody but does not state the fixative (A01678 tissue-IHC caption). Record the actual fixative, fixation duration and processing history for each block before comparing staining across samples (standard IHC practice). Run adjacent sections through the same retrieval and chromogenic detection procedure, changing only the retrieval condition if an established block stains poorly (standard IHC practice). Interpret a negative result against a processed positive control and intact tissue morphology; neither alone establishes that fixation removed the SREBF2 epitope (standard IHC practice).
Why do I see cytoplasmic SREBF2 when I expected nuclear staining?
SREBF2 can be present at the endoplasmic reticulum, Golgi and COPII-coated vesicle membranes, while its processed transcription-factor form enters the nucleus (UniProt Q12772 subcellular location and processing). Assess nuclear staining against the counterstain and record cytoplasmic staining separately within the same cell population (standard IHC practice). Membrane-associated signal can be biologically plausible, but diffuse chromogen over every compartment warrants a no-primary control and inspection for precipitate or section folds (UniProt Q12772 topology; standard IHC practice). HPA reports nuclear expression in most tissues, so a wholly cytoplasmic pattern should prompt review of controls, retrieval and antibody epitope before biological interpretation (HPA tissue IHC).
Can the antibody distinguish precursor SREBF2 from its processed form in tissue?
Do not infer form specificity without a mapped epitope for the catalog antibody (standard antibody validation practice). SREBF2 has 2 annotated isoforms, a full-length 1–1141 chain and a processed 1–484 chain (UniProt Q12772 isoforms and processing). An epitope within the processed N-terminal sequence could recognize both forms, whereas an epitope beyond residue 484 would not recognize that processed chain (UniProt Q12772 processing; epitope-dependent inference). Compare nuclear and extranuclear staining with independently validated controls, and report the observed compartment pattern without assigning an isoform or cleavage state from chromogen alone (standard IHC practice).
How can I examine SREBF2 localisation by IF alongside a cell-type marker?
On the separate IF/ICC workflow, multiplex SREBF2 with a validated lymphoid-cell marker when examining appendix lymphoid tissue, where HPA reports high SREBF2 staining (HPA tissue IHC; standard IF practice). Choose a far-red fluorophore for the weaker channel and check unstained tissue at each wavelength to identify autofluorescence before setting exposure (standard IF practice). Select permeabilisation according to the antibody epitope: N-terminal residues 1–479 and C-terminal residues 555–1139 face the cytoplasm, while residues 501–533 are lumenal (UniProt Q12772 topology). Confirm any nuclear overlap with a nuclear counterstain and single-stain controls; HPA supports nucleoplasmic, vesicular and additional endoplasmic-reticulum localisation (HPA subcellular; standard IF practice).
What should I check when SREBF2 chromogen appears across the whole section?
Inspect a no-primary section for endogenous enzyme signal, nonspecific secondary-antibody binding and chromogen precipitate before changing the catalog antibody concentration (standard chromogenic IHC practice). Include an endogenous peroxidase block before DAB detection when using an HRP system, and check whether background persists in the no-primary control (standard chromogenic IHC practice). Titrate the primary antibody around the documented 5 µg/mL spleen-image concentration only as an experiment, since that caption supplies no fixation or retrieval details (A01678 tissue-IHC caption). Compare staining in target cells with surrounding structures and avoid scoring folds, tissue edges or pigment as intracellular SREBF2 (standard IHC practice).
How should I quantify nuclear SREBF2 staining across tissue samples? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and tissue region before scoring, then count only intact cells with interpretable nuclei (standard IHC quantification practice). Report the percentage of nuclear-positive cells and staining intensity separately, or calculate an H-score from intensity categories 0–3 for a possible range of 0–300 (standard IHC quantification practice). Normalise counts to the number of eligible cells in the same compartment, or report positive-cell density per mm² of viable tissue when cell counts are impractical (standard IHC quantification practice). Keep cytoplasmic staining as a separate measure because precursor and processed SREBF2 occupy different compartments (UniProt Q12772 processing and subcellular location).
How do I distinguish meaningful SREBF2 staining from tissue artefact?
A credible result follows cell boundaries and shows reproducible nuclear or membrane-associated intracellular staining, consistent with SREBF2 processing and localisation (UniProt Q12772 processing and subcellular location; standard IHC practice). Compare the expected cell population with controls: HPA reports high staining in appendix lymphoid tissue but no detection in cerebellar granular-layer cells (HPA tissue IHC). Exclude signal confined to cut edges, necrotic areas, folds or a no-primary section, and investigate residual endogenous peroxidase when DAB appears without primary antibody (standard chromogenic IHC practice). Treat apparent differences cautiously because HPA labels its tissue staining approved yet reports low consistency with RNA expression (HPA tissue IHC).
Boster reagents
Best SREBF2 / Sterol regulatory element-binding protein 2 IHC Antibodies
A01678 has IHC and IF images from human spleen (catalog: IHC and IF image captions); Human and Mouse reactivity is listed (catalog: reactivity).
Real IHC data
Anti-SREBF2 Antibody
Cat # A01678
A01678 is listed for IHC-P and IF, with Human and Mouse reactivity (catalog: applications and reactivity). Its IHC and IF images show human spleen at 5 μg/mL and 20 μg/mL, respectively (catalog: IHC and IF image captions).
Which to pick: Choose A01678 for paraffin-section IHC (catalog: IHC-P); its human spleen IHC caption reports 5 μg/mL but does not report the fixative (catalog: IHC image caption). A01678 is also the IF choice (catalog: IF application and human spleen IF image caption); ICC validation is unreported (catalog: applications). For Mouse samples, A01678 has listed reactivity but no Mouse IHC or IF image in the payload; its host is Rabbit and clone type is unreported (catalog: reactivity, image captions, host, clone).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q12772 (SRBP2_HUMAN, Sterol regulatory element-binding protein 2).
- Human Protein Atlas. SREBF2 tissue IHC expression (reliability: Approved).
- Human Protein Atlas. SREBF2 subcellular location (ICC-IF): Mainly localized to the nucleoplasm and vesicles. In addition localized to the endoplasmic reticulum..
- Human Protein Atlas. SREBF2 antibody validation summary (2 antibodies).
- Integrative analysis of cholesterol homeostasis reveals prognostic subtypes and immune features in clear cell renal cell carcinoma. Discover oncology 2025 — PMC12227387.
- Investigating the Diagnostic and Therapeutic Potential of SREBF2-Related Lipid Metabolism Genes in Colon Cancer. OncoTargets and therapy 2023 — PMC10723182.
- SREBF2 enhances lipid metabolism and represses anti-tumor immune responses in cervical cancer by increasing ACAT2. Communications biology 2026 — PMC12992596.
- A survival prediction model and nomogram based on immune-related gene expression in chronic lymphocytic leukemia cells. Frontiers in medicine 2022 — PMC9806429.
- PubMed PMID:7903453 — UniProt-cited evidence.
- PubMed PMID:15461802 — UniProt-cited evidence.
- PubMed PMID:10591208 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for SREBF2 (Sterol regulatory element-binding protein 2). Expected localisation Nuclear staining in most tissues (HPA tissue IHC); ER/Golgi precursor (UniProt). Antigen retrieval: Tris-EDTA pH 9.0 HIER, 95–98 °C, 20 min (rule: nuclear antigen). Validated IHC antibody: A01678 at 5 μg/mL (the product's own IHC validation image is shown). Controls (HPA): positive Appendix (Lymphoid tissue); negative Cerebellum. IHC workflow; specimen preparation unspecified. Protocol steps: 1. Sample: Tissue sections; selected-image fixative not specified (standard IHC workflow); 2. Fixation: Image fixative and duration unreported (datasheet A01678); 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-induced epitope retrieval in Tris-EDTA buffer, pH 9.0 , 20 min at 95–98 °C…; 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% normal serum of the secondary host, 30 min , room temperature (standard); 8. Primary antibody: Rabbit anti-SREBF2, 5 μg/mL (datasheet A01678); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A01678); Caveat: Sterol status can change processing and nuclear signal (UniProt); Isoform / epitope: 2 isoforms; processed 1–484 lacks C-terminal epitopes (UniProt); Regulation: Low sterols promote nuclear SREBF2 (UniProt). Evidence: UniProt Q12772, HPA tissue IHC · Approved, HPA ICC-IF · Mainly localized to the nucleoplasm and vesicles. In addition…, Datasheet A01678, PMC10723182, PMC12992596. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/SREBF2/immunohistochemistry-ihc-guide.