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SREBF2 / Sterol regulatory element-binding protein 2 · IHC design guide
Design Immunohistochemistry for SREBF2
Plan SREBF2 IHC-P around nuclear staining in most tissues (HPA tissue IHC). Use colon glandular cells as a positive reference (HPA tissue IHC) and start the catalog antibody at 5 μg/mL (datasheet).
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) | |
| Staining pattern | Nuclear signal in most tissues, including colon glands (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 | Keep fixation consistent across sections. (standard IHC practice; not target-specific) | |
| Caveat | Sterol status can shift ER versus nuclear localisation (UniProt) | |
| Regulation | Low sterols promote nuclear translocation (UniProt) | |
| Isoform / epitope | 2 isoforms; cleaved 1–484 retains N-terminal epitopes (UniProt) |
Section 1
Recommended SREBF2 IHC & IF Protocols
The catalog antibody’s IHC-P protocol is followed by a published SREBF2 protocol for paraffin-embedded tumor sections (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 HIER at 95–98 °C for 20 min (page retrieval); the published protocol reports 10 min of retrieval without specifying buffer or temperature (PMC12992596).
Section 2
What Is the Expected SREBF2 Staining Pattern?
SREBF2 IHC is predominantly nuclear in most tissues (HPA tissue IHC: Approved; low consistency with RNA expression). High staining is reported in colon glandular cells, esophageal squamous epithelial cells and placental trophoblastic cells (HPA tissue IHC: High). Its membrane-bound precursor occupies the ER and Golgi, while the processed transcription factor enters the nucleus (UniProt Q12772: topology, processing and subcellular location).
| Distinct nuclear staining in colon glandular cells or placental trophoblastic cells (HPA tissue IHC: High). | This matches the reported tissue pattern and the nuclear destination of processed SREBF2 (HPA tissue IHC: nuclear expression in most tissues; UniProt Q12772: processing). Compare intensity with an appropriate control on the same run (standard IHC practice). |
| Only diffuse, uniform cytoplasmic staining, with no distinct nuclear or structured membrane-associated signal. | Question this pattern: HPA reports predominantly nuclear tissue staining, while UniProt also places the precursor at membranes (HPA tissue IHC; UniProt Q12772: subcellular location). A cytoplasmic signal alone does not identify an artefact; review controls and staining morphology (standard IHC practice). |
| Strong staining in cells reported as undetected, such as cardiomyocytes or pancreatic exocrine glandular cells (HPA tissue IHC: Not detected). | This conflicts with those cell-specific HPA observations and warrants investigation for cross-reactivity or endogenous detection activity (HPA tissue IHC; standard IHC practice). Do not treat an entire heart or pancreas section as a negative control based on those cell-level entries. |
| Color covers tissue edges, empty areas or many cell types without a clear cellular pattern. | Diffuse background can obscure the nuclear pattern reported for SREBF2 (HPA tissue IHC: nuclear expression in most tissues). Check the no-primary control and detection reagents for nonspecific or endogenous signal (standard IHC practice). |
| No nuclear signal in colon glandular cells despite an otherwise interpretable section (HPA tissue IHC: High). | The result is discordant with the reported example, but HPA rates tissue IHC Approved with low consistency against RNA expression (HPA tissue IHC). Check run controls and assay conditions before interpreting the section as biologically negative (standard IHC practice). |
💡Expected SREBF2 appearanceA convincing IHC positive shows distinct nuclear staining in reported high-staining cells such as colon glandular cells; diffuse color without cellular definition is suspect (HPA tissue IHC: nuclear expression; High in colon glandular cells; standard IHC practice).
| Tissue and cell selection | Use the named cell population when judging a control: esophageal squamous epithelial cells are High, whereas cardiomyocytes are Not detected (HPA tissue IHC). HPA's calls describe observed staining, with low consistency against RNA expression (HPA tissue IHC: Approved). |
| Sterol-dependent location | At high sterol levels, the SCAP–SREBF2 complex is retained in the ER; low sterol promotes Golgi transport, processing and nuclear entry (UniProt Q12772: subcellular location and function). Nuclear and membrane-associated patterns can therefore reflect different forms; IHC alone does not measure sterol state (UniProt Q12772; standard IHC interpretation). |
| Antibody epitope and processed form | The precursor spans residues 1–1141 and the processed form residues 1–484 (UniProt Q12772: processing). Whether a particular antibody detects both depends on its epitope, which is unspecified here; avoid assigning a signal to one form without epitope information (UniProt Q12772: processing; standard IHC interpretation). |
| Antibody validation | HPA031962 is Approved for IHC; HPA031963 is Supported for ICC, with no IHC status supplied for that antibody (HPA antibodies). Apply each validation status only to its stated application; it does not establish equivalent performance for another antibody (HPA antibodies). |
| What should IF/ICC show? | HPA reports mainly nucleoplasm and vesicles, with additional ER localization; all three locations are marked Supported (HPA subcellular ICC-IF). Use the separate IF/ICC guide for that application; this tissue IHC pattern does not supply an IF/ICC protocol (HPA subcellular ICC-IF; HPA tissue IHC). |
| Situation | Likely cause | Next action |
|---|---|---|
| Colon glandular cells show no convincing nuclear staining (HPA tissue IHC: High). | A failed staining run or an unsuitable assay setting is possible; a single negative section cannot establish which (standard IHC practice). | Review a known-positive control, the antibody's IHC validation, antigen retrieval conditions and detection steps; repeat after identifying a failed control or step (HPA antibodies: HPA031962 IHC Approved; standard IHC practice). |
| Cardiomyocytes stain strongly despite the reported absence of signal (HPA tissue IHC: Not detected). | Cross-reactivity or endogenous detection activity may produce misleading color (standard IHC practice). | Compare a no-primary control and inspect whether staining follows cell boundaries or background; reassess specificity before scoring cardiomyocytes positive (standard IHC practice). |
| Color is widespread but nuclei cannot be distinguished. | Excess background or counterstain can hide the predominantly nuclear tissue pattern (HPA tissue IHC; standard IHC practice). | Inspect no-primary and counterstain controls; adjust blocking, detection or counterstain using the assay's validated workflow, then reassess nuclear definition (standard IHC practice). |
| Signal appears vesicular or ER-associated rather than predominantly nuclear. | The membrane-bound precursor and processed nuclear form have different locations (UniProt Q12772: topology and processing); HPA also supports vesicle and ER signal in ICC-IF (HPA subcellular ICC-IF). | Document the compartment, review antibody epitope information if available and compare tissue nuclear staining with controls; do not call structured membrane signal nonspecific solely because it is extranuclear (UniProt Q12772; standard IHC practice). |
| Two antibodies yield different tissue patterns. | The supplied HPA statuses cover different applications: HPA031962 is IHC Approved, while HPA031963 is ICC Supported (HPA antibodies). Their staining cannot be assumed interchangeable. | Check each antibody's application validation and epitope information before comparing results; score tissue IHC against cell-specific HPA observations with the stated reliability caveat (HPA antibodies; HPA tissue IHC). |
| A negative call is based on an entire tissue containing a reported undetected cell type. | HPA's absence calls identify specific cells, such as pancreatic exocrine glandular cells and liver cholangiocytes, rather than every cell in those organs (HPA tissue IHC: Not detected). | Score the named cell population separately and check section quality and run controls before assigning a negative result (HPA tissue IHC; standard 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 cardiomyocytes as the negative tissue (HPA: Not detected in cardiomyocytes); on the positive slide, assess adjacent nonglandular cells for background-level staining, but do not assume they are true negatives because SREBF2 is ubiquitously expressed (UniProt: tissue specificity).
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 a no-primary, secondary-only control and an isotype control matched to the primary antibody’s host species, clonality and concentration; use a matched SREBF2-knockout specimen as a biological negative if available (standard IHC practice). In colon sections, block endogenous peroxidase and inspect gland lumens for trapped chromogen (standard IHC practice).
⚠️Feasibility: A target-specific fixation window and retrieval effect are unreported in the supplied evidence; the A01678 spleen IHC caption gives 5 µg/mL but does not report the fixative (selected tissue-IHC caption). Optimize antigen retrieval empirically for paraffin sections; the supplied evidence does not establish whether frozen sections or IF are easier, although ICC-IF images exist for this target (HPA: subcellular ICC-IF). Colon luminal mucus can complicate interpretation of chromogenic staining (standard IHC practice).
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
Use compartment-aware controls and consistent section processing to troubleshoot SREBF2 staining in chromogenic paraffin IHC (UniProt Q12772; standard IHC practice).
What retrieval conditions should I try when SREBF2 staining is weak?
For paraffin IHC, use Tris-EDTA pH 9.0 HIER at 95–98 °C for 20 min (page retrieval setting: nuclear antigen). Keep heating, cooling, and section thickness consistent across the comparison, then inspect nuclear staining and tissue preservation together (standard IHC practice; HPA: nuclear expression in most tissues). If staining remains weak, compare a cautiously shortened retrieval with the specified 20 min condition before increasing antibody concentration, because excessive heating can damage morphology (standard IHC practice). Include a known staining tissue section in the same run and interpret weak signal in light of SREBF2 processing and sterol dependent nuclear entry (HPA: tissue IHC; UniProt Q12772 processing).
Could fixation explain inconsistent SREBF2 staining between paraffin sections?
The selected SREBF2 tissue IHC caption reports human spleen staining at 5 µg/mL but does not state its fixative (A01678 tissue IHC caption). Target specific fixation sensitivity is therefore unknown; neither tissue staining patterns nor SREBF2 topology establishes how fixation changes this antibody’s signal (A01678 tissue IHC caption; HPA: tissue IHC; UniProt Q12772 topology). Compare sections processed with documented, matched fixation and the same Tris-EDTA pH 9.0, 20 min retrieval before attributing a difference to biology (page retrieval setting; standard IHC practice). Record fixation history, section age, morphology, and control staining for each batch so an apparent loss of nuclear signal can be investigated (standard IHC practice; HPA: nuclear expression in most tissues).
How should I interpret nuclear versus cytoplasmic SREBF2 staining?
In chromogenic IHC, score nuclear and cytoplasmic SREBF2 separately because the processed 1–484 chain enters the nucleus while precursor SREBF2 resides in the endoplasmic reticulum membrane (UniProt Q12772 processing and localisation). Low sterol conditions promote transport through COPII vesicles to the Golgi and subsequent processing, whereas high sterol conditions retain the SCAP–SREBF complex in the endoplasmic reticulum (UniProt Q12772). Nuclear staining in many tissues is plausible, but a cytoplasmic signal alone does not measure transcriptional activity (HPA: nuclear expression in most tissues; UniProt Q12772 function). Compare compartment patterns within intact cells and matched staining runs, using hematoxylin to identify nuclei and section morphology (standard IHC practice).
Can this antibody distinguish precursor, processed SREBF2, and its isoforms?
Check the catalog antibody’s documented immunogen before assigning a chromogenic pattern to a particular SREBF2 form; the supplied caption gives a concentration of 5 µg/mL, not an epitope (A01678 tissue IHC caption). SREBF2 has 2 listed isoforms and a processed 1–484 chain, while its membrane spans lie at residues 480–500 and 534–554 (UniProt Q12772 isoforms, processing and topology). An antibody recognizing a shared N-terminal region could detect precursor and processed protein, whereas an epitope absent from the processed chain would not report that nuclear form (UniProt Q12772 processing; standard antibody interpretation). Treat nuclear staining as evidence of the processed form only after epitope mapping and appropriate controls support that assignment (UniProt Q12772 processing; standard IHC practice).
How can I investigate a conflicting SREBF2 IF result alongside my IHC finding?
Use IF as a separate localisation check, pairing SREBF2 with a marker for the cell population scored in IHC, such as a lymphoid marker for appendix lymphoid tissue (HPA: high in appendix lymphoid tissue; standard IF practice). Select spectrally separated fluorophores and inspect an unstained section in each channel before choosing a signal channel above tissue autofluorescence (standard IF practice). Choose permeabilisation after checking the antibody epitope: residues 1–479 and 555–1139 face the cytoplasm, while 501–533 is lumenal (UniProt Q12772 topology; standard IF practice). Compare nuclear, vesicular, and endoplasmic reticulum patterns with the compartment resolved IHC result, without assuming the IHC caption establishes IF performance (HPA: supported subcellular locations; A01678 tissue IHC caption).
What should I check when SREBF2 DAB staining is diffuse or widespread?
Start by comparing the stained section with a no-primary control, then check whether diffuse DAB persists after the routine peroxidase block (standard chromogenic IHC practice). Inspect tissue edges, folds, damaged areas, and red blood cell rich regions before calling broad brown signal cellular SREBF2 staining (standard IHC practice). If background rises with primary antibody concentration, titrate below the caption’s 5 µg/mL test point while keeping retrieval at Tris-EDTA pH 9.0 for 20 min (A01678 tissue IHC caption; page retrieval setting; standard IHC practice). Evaluate nuclei separately from diffuse cytoplasm because HPA reports nuclear expression in most tissues and UniProt also places precursor protein in membrane compartments (HPA: tissue IHC; UniProt Q12772 localisation).
How should I quantify SREBF2 staining across tissue groups? ⚠ ANSWER MARKED FOR VERIFICATION
For paraffin IHC, define the scored cell population and compartment before analysis, then report nuclear H-score or percent positive cells with the threshold held constant across slides (standard IHC practice; HPA: nuclear expression in most tissues). Record cytoplasmic staining separately because SREBF2 precursor occupies membrane compartments and processing releases a nuclear transcription factor (UniProt Q12772 processing and localisation). Normalise positive counts to the number of intact cells of the same type, or report positive cell density per mm² of viable tissue when cell boundaries cannot be resolved (standard IHC practice). Apply the same exposure or scan settings, counterstain review, and exclusion rules for folds and necrosis to every group (standard IHC practice).
What makes an apparent SREBF2-positive cell credible rather than artefactual?
A credible IHC positive has interpretable cellular boundaries and a compartment pattern consistent with SREBF2 biology: nuclear processed protein or precursor associated with intracellular membranes (UniProt Q12772 processing and localisation; standard IHC practice). Compare the observed cell type with tissue context; HPA reports high staining in bone marrow hematopoietic cells but no detection in heart cardiomyocytes, with low consistency between staining and RNA data overall (HPA: tissue IHC and reliability description). Reject isolated edge staining, necrotic debris, or signal retained in the no-primary control as evidence of specific expression (standard IHC practice). Where brown deposits follow endogenous enzyme activity, verify the peroxidase block and repeat the comparison with matched controls before interpreting a biological difference (standard chromogenic IHC practice).
Boster reagents
Best SREBF2 / Sterol regulatory element-binding protein 2 IHC Antibodies
A01678 is an IHC-P and IF antibody reactive with human and mouse SREBF2 (catalog: applications and reactivity), with human spleen IHC and IF images (catalog: image captions).
Real IHC data
Anti-SREBF2 Antibody
Cat # A01678
A01678 has an IHC image of human spleen at 5 μg/mL (catalog: IHC image caption). A01678 also has an IF image of human spleen at 20 μg/mL (catalog: IF image caption); mouse reactivity is listed without a mouse image (catalog: reactivity and image captions).
Which to pick: Choose A01678 for paraffin-section IHC: IHC-P is listed, and its own IHC image shows human spleen; the fixative is unreported (catalog: applications and IHC image caption). For IF, A01678 has a human spleen image; ICC is not listed as an application (catalog: IF image caption and applications). For mouse samples, A01678 lists mouse reactivity, but the supplied IHC and IF images show human spleen only (catalog: reactivity and image captions).
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). 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: Keep fixation consistent across sections. (standard IHC practice; not target-specific); Caveat: Sterol status can shift ER versus nuclear localisation (UniProt); Isoform / epitope: 2 isoforms; cleaved 1–484 retains N-terminal epitopes (UniProt); Regulation: Low sterols promote nuclear translocation (UniProt). Evidence: UniProt Q12772, HPA tissue IHC · Approved, HPA ICC-IF · Mainly localized to the nucleoplasm and vesicles. In addition…, Datasheet A01678, PMC12992596. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/SREBF2/immunohistochemistry-ihc-guide.