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CHD2 / ATP-dependent chromatin remodeler CHD2 · IHC design guide
Design Immunohistochemistry for CHD2
CHD2 shows variable nuclear staining across most tissue types (HPA tissue IHC). This IHC-P guide uses the catalog antibody and tissue evidence to help plan controls and interpret staining.
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
CHD2 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 tissue sections (HPA tissue IHC) | |
| Staining pattern | Variable nuclear staining across most tissue types (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6 HIER, heat-mediated (datasheet A04079) | |
| Positive control | Cerebellum+4 more · see all | |
| Negative control | Adipose tissue+1 more · see all |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep fixation conditions consistent across sections. (standard IHC practice; not target-specific) | |
| Caveat | Skeletal myocytes: not detected (HPA tissue IHC) | |
| Regulation | Staining intensity varies by tissue (HPA tissue IHC) | |
| Isoform / epitope | 3 isoforms; check epitope coverage (UniProt) |
Section 1
Recommended CHD2 IHC & IF Protocols
The catalog antibody’s IHC-P protocol (datasheet: A04079) is paired with a published CHD2 staining protocol for mouse testis sections (PMC9700024).
| Sample | Paraffin-embedded rat brain tissue; fixative not specified (datasheet A04079) |
| Fixation | Image fixative and duration unreported (datasheet A04079); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat retrieval: Citrate pH 6, 20 min (datasheet A04079) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% goat serum (datasheet A04079) |
| Primary antibody | Rabbit anti-CHD2, 0.5-1μg/ml (datasheet A04079) |
| Primary incubation | Overnight at 4 °C (datasheet A04079) |
| Detection | Streptavidin-biotin complex (SABC), DAB chromogen (datasheet A04079) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | CHD2-positive staining in cells in granular layer of cerebellum (HPA tissue IHC: High). HPA tissue profile: Nuclear expression at variable levels in most tissue types. No signal in the no-primary control. |
💡Decision noteStart with heat-mediated citrate pH 6 retrieval (datasheet: A04079); the published mouse testis excerpt does not specify retrieval conditions (PMC9700024).
Section 2
What Is the Expected CHD2 Staining Pattern?
In paraffin-section IHC, expect CHD2 predominantly in nuclei, at variable levels across most tissues (HPA tissue IHC: Enhanced; medium consistency with RNA). Strong examples include cerebellar granular-layer cells, cortical and hippocampal neurons, and endometrial glandular cells (HPA tissue IHC: High). A nuclear pattern agrees with its annotated location and lack of a transmembrane segment (UniProt O14647: nucleus; topology).
| Distinct nuclear staining in cerebellar granular-layer cells or cortical neurons, with less staining in adjacent cells. | This fits the expected compartment and two reported High cell populations (UniProt O14647: nucleus; HPA tissue IHC: High). Compare nuclei within the same section: HPA describes variable expression, so uniform intensity across every cell is unnecessary (HPA tissue IHC: variable nuclear expression). |
| Strong, widespread cytoplasmic staining obscures nuclei in a paraffin section. | Treat this as a questionable IHC pattern because HPA reports nuclear tissue staining and UniProt places CHD2 in the nucleus (HPA tissue IHC: nuclear expression; UniProt O14647: nucleus). Check background and antibody controls before calling the cytoplasmic signal CHD2. ICC-IF has a separate localisation record (HPA subcellular ICC-IF: additional locations). |
| Adipocytes or skeletal-muscle myocytes stain strongly while nearby expected nuclei are weak. | Those cell types were Not detected in HPA tissue IHC; strong signal there raises concern for cross-reactivity or endogenous chromogenic activity (HPA tissue IHC: adipocytes and myocytes Not detected; general IHC practice). Judge the named cell types, since these records do not declare every cell in either tissue negative. |
| Colour spreads across stroma, extracellular spaces, or most cells without clear nuclear boundaries. | A diffuse deposit cannot establish CHD2-positive cells; assess nonspecific antibody binding, endogenous detection activity, and wash adequacy with suitable controls (general IHC practice). Compare the distribution with HPA's variable nuclear pattern rather than assigning a score to the entire field (HPA tissue IHC: nuclear expression). |
| No nuclear signal appears in an HPA High population, such as endometrial glandular cells. | A negative result there calls for a technical check of staining and tissue integrity; it does not by itself prove CHD2 absence (HPA tissue IHC: endometrial glandular cells High; general IHC practice). HPA's Enhanced tissue assessment still notes medium consistency with RNA, so avoid treating one reference image as an absolute threshold (HPA tissue IHC: reliability). |
💡Expected CHD2 appearanceCall a positive IHC result when identifiable nuclei stain in a reported High population, such as hippocampal neurons or testicular Sertoli cells (HPA tissue IHC: High; UniProt O14647: nucleus); strong diffuse extranuclear colour or staining confined to adipocytes is suspect (HPA tissue IHC: adipocytes Not detected; general IHC practice).
| Tissue and cell selection | HPA reports variable nuclear expression across most tissues, with High signal in several neuronal, glandular, and Sertoli populations (HPA tissue IHC). Select a named cell population for scoring; a mixed section can contain cells with different staining levels. |
| Reference antibody validation | The HPA tissue profile is Enhanced, while its antibody entries differ: HPA060744 is IHC Enhanced and HPA060960 is IHC Supported (HPA antibodies; HPA tissue IHC). These labels support interpretation of the reference pattern; they do not validate every catalog antibody. |
| IF/ICC localisation Q&A | What should IF/ICC show? Mainly nucleoplasmic signal, with reported additional nucleolar, Golgi-apparatus, and vesicle localisation (HPA subcellular ICC-IF: approved locations). Those ICC-IF observations do not establish an equally prominent extranuclear pattern in paraffin-section IHC. |
| Isoforms and epitope coverage | UniProt lists 3 CHD2 isoforms (UniProt O14647: isoforms). If a tested antibody's epitope differs among them, staining could reflect its coverage; no epitope or isoform-specific IHC performance is supplied, so do not assign an observed pattern to one isoform. |
| Retrieval and detection context | Antigen retrieval and chromogenic detection are general paraffin-IHC workflow variables (general IHC practice). No supplied CHD2 source establishes fixation sensitivity or an optimal retrieval condition; use the IHC-validated antibody's stated conditions, then evaluate controls. |
| Situation | Likely cause | Next action |
|---|---|---|
| No stain in cerebellar granular-layer cells or another HPA High population. | The run may have insufficient detection or a tissue-quality problem; the image alone cannot identify which (HPA tissue IHC: High; general IHC practice). | Check the positive-control section, reagent sequence, and whether nuclei remain interpretable; review the IHC-validated antibody's stated conditions before adjusting retrieval or dilution (general IHC practice). |
| Nuclei are visible, but colour is mainly cytoplasmic throughout the section. | The distribution conflicts with the reported tissue-IHC pattern; nonspecific binding or detection background is possible (HPA tissue IHC: nuclear expression; general IHC practice). | Compare a matched negative control and a reported High cell population, then reassess blocking, washing, and detection conditions (HPA tissue IHC: High populations; general IHC practice). |
| Adipocytes or skeletal-muscle myocytes look strongly positive. | HPA reports CHD2 as Not detected in those named cell types; cross-reactivity or endogenous chromogenic activity may account for colour (HPA tissue IHC; general IHC practice). | Check controls for antibody-independent colour and compare nuclear staining in a reported High population; score each cell type separately (general IHC practice; HPA tissue IHC: High populations). |
| The whole section has diffuse colour and cell boundaries are hard to resolve. | Excess background can obscure a variable nuclear pattern; its source cannot be determined from the test section alone (HPA tissue IHC: variable nuclear expression; general IHC practice). | Inspect a matched negative control, confirm wash and blocking steps, and reassess chromogen development using the IHC-validated antibody's conditions (general IHC practice). |
| Signal differs sharply between cell types within one otherwise interpretable section. | Cell-dependent variation can be genuine: HPA reports High, Medium, Low, and Not detected populations (HPA tissue IHC). | Identify and score the named cell populations separately; compare their nuclear pattern with HPA tissue observations before changing the staining workflow (HPA tissue IHC: variable nuclear expression). |
| An IF/ICC image shows nucleolar or punctate extranuclear signal alongside nuclear staining. | HPA reports additional nucleolar, Golgi-apparatus, and vesicle locations in ICC-IF (HPA subcellular ICC-IF: approved locations). | Interpret that image using the separate IF/ICC guide and its controls; for paraffin IHC, retain the reported variable nuclear tissue pattern as the reference (HPA subcellular ICC-IF; HPA tissue IHC). |
Experiment evidence
IHC & IF images showing CHD2 staining
A04079 · IHCIHC analysis of CHD2 using anti-CHD2 antibody (A04079). CHD2 was detected in paraffin-embedded section of rat brain tissue . Heat mediated a
A04079 · IHCIHC analysis of CHD2 using anti-CHD2 antibody (A04079). CHD2 was detected in paraffin-embedded section of mouse brain tissue . Heat mediated
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000173575-CHD2/tissue/cerebral+cortex#imid_16832467
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000173575-CHD2/tissue/colon#imid_16832385
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000173575-CHD2/tissue/liver#imid_16832386
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000173575-CHD2/tissue/kidney#imid_16832379
HPA · ICC-IF Rh30Human Protein Atlas immunofluorescence, Rh30, antibody HPA060744; green = CHD2, blue = nucleus, red = microtubules.
HPA · ICC-IF Rh30Human Protein Atlas immunofluorescence, Rh30, antibody HPA060744; green = CHD2, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for CHD2 IHC & IF
🧪Run cerebellum first: cells in the granular layer should stain (HPA: High). Use adipose tissue adipocytes as the negative tissue (HPA: Not detected); HPA does not identify an internal negative cell population in cerebellum, so cells without nuclear signal on that slide should be treated as provisional background references rather than confirmed biological negatives (HPA: cerebellum row; UniProt: nucleus).
Positive control tissue: Cerebellum (Cells in granular layer, HPA High)
Negative control tissue: Adipose tissue (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show CHD2 in Rh30, SiHa, U2OS, RPTEC/TERT1, hTERT-RPE1 (serum starved), with annotated localisation: Nucleoplasm (approved) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control, a rabbit host- and isotype-matched control appropriate to the primary’s clonality, and CHD2-knockout material as a biological negative (caption: rabbit primary; standard IHC control practice). For cerebellar DAB staining, block endogenous peroxidase and assess endogenous biotin background because the reported detection uses a streptavidin–biotin complex (caption: SABC with DAB; standard IHC practice).
⚠️Feasibility: The selected A04079 paraffin-section caption does not state the fixative, and the supplied evidence reports no CHD2-specific fixation window or fixation effect (caption: fixative unreported). Heat retrieval in citrate buffer at pH 6 for 20 minutes was used in the reported rat brain IHC example, but a retrieval dependency has not been established (caption: retrieval conditions). The evidence does not establish whether frozen sections or IF are easier; brain autofluorescence can complicate IF interpretation (standard IF practice).
HPA tissue IHC evidence for CHD2
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Enhanced — 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 |
|---|---|---|---|---|
| Cerebellum | Cells in granular layer | High | Protein (IHC) | HPA → |
| Cerebral cortex | Neuronal cells | High | Protein (IHC) | HPA → |
| Endometrium | Glandular cells | High | Protein (IHC) | HPA → |
| Epididymis | Glandular cells | High | Protein (IHC) | HPA → |
| Hippocampus | Neuronal cells | High | Protein (IHC) | HPA → |
Section 3
Advanced CHD2 IHC Tips
Troubleshoot CHD2 staining in paraffin sections by checking nuclear localisation, retrieval, cell context and controls before comparing chromogenic signal across samples.
How should I retrieve CHD2 antigen in paraffin sections?
Start with heat-mediated citrate retrieval at pH 6 for 20 minutes (datasheet A04079). The catalog antibody produced CHD2 staining in a paraffin-embedded rat brain section after that treatment; the caption does not state the fixative (datasheet A04079). If nuclear staining is weak, compare a modestly longer heating step or an alternative retrieval buffer on adjacent sections, while keeping antibody concentration and detection constant (standard IHC practice). Judge the result by nuclear signal, tissue preservation and background together, using a no-primary control and the reported high neuronal staining in cerebral cortex as a reference (UniProt O14647 localisation; HPA: High in cerebral cortex neuronal cells; standard IHC practice).
Could fixation explain weak or uneven CHD2 staining?
Target-specific fixation sensitivity is unknown: the selected paraffin-section caption does not state its fixative (datasheet A04079). Record fixative, fixation duration, tissue thickness and processing history for each section, then compare similarly processed samples under the same pH 6 citrate retrieval conditions (datasheet A04079; standard IHC practice). Where fixation differs, assess nuclear signal alongside morphology and a no-primary control before adjusting antibody concentration (UniProt O14647 localisation; standard IHC practice). Do not assign weak staining to CHD2-specific fixation damage without a controlled comparison; regional underfixation, overfixation or uneven processing can also change chromogenic staining (standard IHC practice).
Where should convincing CHD2 staining appear in tissue?
Prioritise staining in nuclei because CHD2 is annotated as nuclear and HPA reports variable nuclear expression across most tissue types (UniProt O14647 localisation; HPA tissue IHC profile). The HPA subcellular dataset places CHD2 mainly in the nucleoplasm, with additional nucleolar, Golgi and vesicular localisation in cell imaging; those additional sites require independent confirmation in tissue (HPA subcellular). Compare nuclear DAB with a light nuclear counterstain and inspect whether signal falls within intact cells rather than section folds or damaged edges (standard IHC practice). Cerebral cortex neuronal cells and cerebellar granular-layer cells provide reported high-expression contexts for assessing the expected pattern (HPA: High in cerebral cortex neuronal cells; HPA: High in cerebellum granular-layer cells).
How can isoforms or epitope accessibility affect CHD2 IHC?
CHD2 has 3 listed isoforms, but the supplied evidence does not map the catalog antibody epitope to an isoform (UniProt O14647 isoforms; datasheet A04079). Confirm the antibody's immunogen or epitope documentation before claiming isoform-specific staining, and keep retrieval and detection matched when comparing samples (standard IHC practice). CHD2 contains two chromo domains and helicase regions, and its record lists multiple phosphorylation sites; neither feature establishes how this antibody stains processed tissue (UniProt O14647 domains and modified residues). If an epitope is later mapped, check whether it is retained in the isoform of interest before interpreting absent nuclear signal as absent CHD2 (UniProt O14647 isoforms; standard IHC practice).
How should I examine CHD2 by multiplex IF?
For a separate IF experiment, pair CHD2 with a validated marker of the cell population being examined and a nuclear stain; HPA reports high CHD2 in cerebral cortex neuronal cells (HPA: High in cerebral cortex neuronal cells; standard IF practice). Choose spectrally separated fluorophores and place the weaker signal in a channel with lower tissue autofluorescence, checking single-stain and no-primary controls (standard IF practice). CHD2 is nuclear and has no transmembrane segment, so use permeabilisation appropriate for access to nuclear epitopes and verify that nuclear morphology remains intact (UniProt O14647 localisation and topology; standard IF practice). Treat IF localisation as complementary evidence; the selected antibody caption documents paraffin-section chromogenic IHC, not an IF protocol (datasheet A04079).
What should I check when CHD2 DAB background is high?
Compare the stained section with a no-primary control to locate background from the secondary reagent, detection chemistry or tissue rather than the primary antibody (standard IHC practice). The documented method used 10% goat serum blocking, 1 μg/ml rabbit primary overnight at 4°C, and a biotin-based DAB detection sequence (datasheet A04079). Check endogenous peroxidase blocking and, for biotin-based detection, endogenous biotin controls as general chromogenic IHC steps (standard IHC practice). If diffuse staining persists, assess washing, primary concentration and DAB development on matched sections while requiring discrete nuclear signal in intact cells (UniProt O14647 localisation; standard IHC practice).
How can I quantify CHD2 staining across tissue sections? ⚠ ANSWER MARKED FOR VERIFICATION
Score CHD2 within prespecified cell populations and tissue regions, using nuclear % positive cells or a nuclear H-score rather than whole-section DAB intensity alone (UniProt O14647 localisation; standard IHC practice). Set a positivity threshold against a no-primary control, then apply the same counterstain, imaging settings and threshold to all compared sections (standard IHC practice). Normalise positive nuclei to the total evaluable nuclei in each annotated population; report the number of fields, sections and biological samples separately (standard IHC practice). Because HPA describes variable nuclear expression across tissues, record cell identity and region with each score before comparing groups (HPA tissue IHC profile; standard IHC practice).
How do I distinguish true CHD2 signal from artefact?
A convincing result shows nuclear staining in morphologically intact cells, consistent with CHD2's annotated nuclear location and HPA's tissue profile (UniProt O14647 localisation; HPA tissue IHC profile). HPA reports high staining in hippocampal neuronal cells and no detection in adipocytes or skeletal-muscle myocytes; use these as context, not absolute pass–fail controls for every specimen (HPA: High in hippocampus neuronal cells; HPA: Not detected in adipocytes and skeletal-muscle myocytes). Distrust signal concentrated at cut edges, folds or necrotic areas, and compare it with no-primary and endogenous-peroxidase controls (standard IHC practice). Predominantly cytoplasmic DAB or staining that tracks tissue damage warrants repeat assessment of retrieval, blocking and morphology before a CHD2-specific conclusion (UniProt O14647 localisation; standard IHC practice).
Boster reagents
Best CHD2 / ATP-dependent chromatin remodeler CHD2 IHC Antibodies
A04079 has IHC images of paraffin-embedded rat, mouse, and human brain sections (A04079 IHC captions). No IF/ICC application or image is listed (catalog applications and IF images).
Real IHC data
Anti-CHD2 Antibody ®
Cat # A04079
A04079 will render with its rat brain IHC figure (A04079 figure tag and image caption). Its additional IHC captions show mouse and human brain sections, while its listed reactivity also includes Monkey (A04079 IHC captions; catalog reactivity).
Which to pick: Choose A04079 for paraffin-section tissue IHC: its captions document rat and mouse brain with citrate retrieval at pH 6, and human brain with EDTA retrieval at pH 8 (A04079 IHC captions). For cross-species work, A04079 lists Human, Monkey, Mouse, and Rat reactivity, though the supplied IHC images cover Human, Mouse, and Rat; the fixative is unreported (catalog reactivity; A04079 IHC captions). No supplied SKU supports an IF/ICC recommendation because A04079 has no listed IF/ICC application, dilution, or image (catalog applications, IF dilution, and IF images).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry O14647 (CHD2_HUMAN, ATP-dependent chromatin remodeler CHD2).
- Human Protein Atlas. CHD2 tissue IHC expression (reliability: Enhanced).
- Human Protein Atlas. CHD2 subcellular location (ICC-IF): Mainly localized to the nucleoplasm. In addition localized to the nucleoli, the Golgi apparatus and vesicles..
- Human Protein Atlas. CHD2 antibody validation summary (2 antibodies).
- Chromodomain helicase DNA-binding domain 2 maintains spermatogonial self-renewal by promoting chromatin accessibility and mRNA stability. iScience 2022 — PMC9700024.
- TDP-43 Promotes Neurodegeneration by Impairing Chromatin Remodeling. Current biology : CB 2017 — PMC5720388.
- CHD2 Regulates Neuron-Glioma Interactions in Pediatric Glioma. Cancer discovery 2024 — PMC11456263.
- PubMed PMID:9326634 — UniProt-cited evidence.
- PubMed PMID:16572171 — UniProt-cited evidence.
- PubMed PMID:15489334 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for CHD2 (ATP-dependent chromatin remodeler CHD2). Expected localisation Nuclear staining in tissue sections (HPA tissue IHC). Antigen retrieval: Citrate pH 6 HIER, heat-mediated (datasheet A04079). Validated IHC antibody: A04079 at 0.5-1μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Cerebellum (Cells in granular layer); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded rat brain tissue; fixative not specified (datasheet A04079); 2. Fixation: Image fixative and duration unreported (datasheet A04079); 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: Citrate pH 6 , 20 min (datasheet A04079); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A04079); 8. Primary antibody: Rabbit anti-CHD2, 0.5-1μg/ml (datasheet A04079); 9. Primary incubation: Overnight at 4 °C (datasheet A04079). Watch for: Fixation: Keep fixation conditions consistent across sections. (standard IHC practice; not target-specific); Caveat: Skeletal myocytes: not detected (HPA tissue IHC); Isoform / epitope: 3 isoforms; check epitope coverage (UniProt); Regulation: Staining intensity varies by tissue (HPA tissue IHC). Evidence: UniProt O14647, HPA tissue IHC · Enhanced, HPA ICC-IF · Mainly localized to the nucleoplasm. In addition localized to the…, Datasheet A04079, PMC9700024. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/CHD2/immunohistochemistry-ihc-guide.