CHMP2B / Charged multivesicular body protein 2b · IHC design guide

Design Immunohistochemistry for CHMP2B

Plan chromogenic CHMP2B IHC in paraffin sections with the catalog antibody at 1:50 (datasheet M01935). Compare cytoplasmic staining by cell type with controls; the tissue IHC profile has uncertain reliability (HPA tissue IHC).

Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use

Immunohistochemistry protocol sheet for CHMP2B (IHC for CHMP2B): expected localisation Cytoplasmic staining in selected tissues (HPA tissue IHC), antibody M01935, validated IHC image, and IHC protocol steps — Printable **CHMP2B** IHC protocol sheet — expected localisation Cytoplasmic staining in selected tissues (HPA tissue IHC), antibody M01935, controls and protocol steps. Open the full CHMP2B IHC guide →

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CHMP2B 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 staining in selected tissues (HPA tissue IHC)
	Staining pattern	Cytoplasmic signal in selected cells, including immune cells (HPA tissue IHC)
	Antigen retrieval	EDTA pH 8.0 HIER, heat-mediated (datasheet M01935)
	Positive control ⓘ	Appendix+4 more · see all
	Negative control ⓘ	Adipose tissue+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	Tissue IHC staining has low RNA concordance (HPA tissue IHC)
	Regulation	Widely expressed; regulation unspecified (UniProt)
	Isoform / epitope	2 isoforms; check epitope coverage (UniProt)

Section 1

Recommended CHMP2B IHC & IF Protocols

The catalog antibody’s IHC-P protocol (datasheet: M01935) is followed by two published CHMP2B paraffin-section protocols (PMC3207829; PMC10758028).

Recommended immunohistochemistry (IHC-P) protocol parameters
Sample	Paraffin-embedded human colon cancer tissue; fixative not specified (datasheet M01935)
Fixation	Image fixative and duration unreported (datasheet M01935); 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 M01935); 20 min, 95–100 °C (standard)
Peroxidase block	3% H2O2, 10 min, room temperature (standard)
Blocking	10% goat serum (datasheet M01935)
Primary antibody	Rabbit monoclonal (clone 25C36) anti-CHMP2B, 1:50 (datasheet M01935)
Primary incubation	Overnight at 4 °C (datasheet M01935)
Detection	HRP-conjugated secondary, DAB chromogen (datasheet M01935)
Counterstain	Hematoxylin, blue, dehydrate and mount (standard)
Expected result	CHMP2B-positive staining in glandular cells of appendix (HPA tissue IHC: Medium). HPA tissue profile: Cytoplasmic expression in selected tissues, including immune cells in several tissues. No signal in the no-primary control.

💡Decision noteStart with EDTA pH 8.0 heat retrieval (datasheet: M01935); the published protocols used distilled water or an unspecified retrieval buffer (PMC3207829; PMC10758028).

Section 2

What Is the Expected CHMP2B Staining Pattern?

CHMP2B is cytosolic and can associate with late endosome membranes; it has no transmembrane segment (UniProt Q9UQN3 topology). In paraffin sections, expect cytoplasmic staining in selected cell populations, including glandular, hematopoietic and tubular cells (HPA: tissue IHC). Interpret intensity cautiously: HPA rates its tissue IHC reliability Uncertain because antibody staining has low consistency with RNA expression (HPA: tissue IHC reliability).

What am I looking at on my slide?
Cytoplasmic staining in colon glandular cells or kidney tubular cells.	This matches HPA's Medium staining in those cell types and UniProt's cytosolic localisation (HPA: tissue IHC; UniProt Q9UQN3 subcellular location). It is a plausible positive pattern, but HPA's Uncertain IHC reliability means the slide alone does not establish specificity (HPA: tissue IHC reliability).
Predominantly nuclear or sharp cell-surface staining, with little cytoplasmic signal.	This conflicts with the reported cytosolic location and absence of a transmembrane segment (HPA: subcellular ICC-IF; UniProt Q9UQN3 topology). Treat it as a possible staining artefact; inspect controls and tissue morphology before assigning CHMP2B localisation (general IHC practice).
Strong staining in adipocytes while neighbouring cells show little signal.	HPA reports adipocytes as Not detected in adipose tissue and breast (HPA: tissue IHC). This mismatch raises cross-reactivity or endogenous detection activity as possibilities; it does not prove either, especially given HPA's Uncertain IHC reliability (HPA: tissue IHC reliability; general IHC practice).
Colour is spread uniformly across tissue, including areas without convincing cellular detail.	A diffuse deposit cannot be assigned confidently to the reported cytoplasmic cell pattern (HPA: tissue IHC; general IHC practice). Review a no-primary control, blocking and chromogen development to distinguish nonspecific staining or detection background from cell-associated signal (general IHC practice).
No staining in colon glandular cells or kidney tubular cells.	Both populations have Medium staining in HPA, so absent signal warrants a workflow check (HPA: tissue IHC). Confirm tissue morphology and detection controls, then review the antibody's validated IHC-P conditions; HPA's uncertain reliability prevents treating either population as a guaranteed positive control (HPA: tissue IHC reliability; general IHC practice).

💡Expected CHMP2B appearanceCall a plausible positive when cytoplasmic staining is discernible in glandular or kidney tubular cells at roughly Medium intensity (HPA: tissue IHC; UniProt Q9UQN3 subcellular location); predominantly nuclear staining or colour without cellular boundaries is suspect (HPA: subcellular ICC-IF; general IHC practice).

How each factor affects the staining
Subcellular location	UniProt places CHMP2B in cytosol and at late endosome membranes, with no transmembrane segment (UniProt Q9UQN3 subcellular location and topology). Score the cellular compartment first; these annotations do not require every endosome to resolve as a separate punctum in chromogenic sections (general IHC practice).
Cell and tissue selection	HPA reports Medium staining in appendix, colon, duodenum and gallbladder glandular cells, bone marrow hematopoietic cells, bronchial epithelium, kidney tubules and hepatocytes (HPA: tissue IHC). Its low tissue RNA specificity and uncertain IHC reliability limit claims that a single positive or negative tissue proves specificity (HPA: tissue IHC).
Validation strength	The tissue IHC assessment is Uncertain, and antibody HPA035069 is also rated Uncertain for IHC (HPA: tissue IHC reliability; HPA: antibodies). Compare cellular pattern and controls before interpreting a new specimen; these ratings do not establish that every observed deposit represents CHMP2B (general IHC practice).
Isoforms and epitope	UniProt lists 2 isoforms but supplies no antibody epitope in this record (UniProt Q9UQN3 isoforms; supplied record). An isoform-specific staining prediction is therefore unsupported; check the selected antibody's epitope information before using staining differences to infer isoform expression (general IHC practice).
Processing and modification	UniProt reports no signal peptide or propeptide, a chain spanning residues 2–213, and modifications at residues 2 and 199 (UniProt Q9UQN3 processing and modified residues). These annotations alone do not establish epitope accessibility or a target-specific fixation effect (supplied record).

Why is my staining missing, weak or wrong?
Situation	Likely cause	Next action
Expected cytoplasmic cells are blank.	Detection may have failed, or conditions for this antibody may be unsuitable; HPA's reference pattern is itself uncertain (HPA: tissue IHC reliability; general IHC practice).	Check tissue preservation and a detection control, then consult the catalog antibody's IHC-P protocol for its specified retrieval and dilution; no such values are supplied here (general IHC practice; supplied record).
Signal is mainly nuclear.	This conflicts with cytosolic localisation and late endosome association (HPA: subcellular ICC-IF; UniProt Q9UQN3 subcellular location).	Inspect a no-primary control, section morphology and counterstain before scoring; if the nuclear pattern persists, treat it as unverified for CHMP2B (general IHC practice).
Adipocytes stain strongly.	HPA marks adipocytes Not detected in adipose tissue and breast; cross-reactivity or endogenous detection activity is possible (HPA: tissue IHC; general IHC practice).	Compare no-primary and detection-only controls, and inspect whether colour follows cells or tissue pigment; avoid calling the adipocyte signal specific on appearance alone (general IHC practice).
The whole section looks hazy.	Diffuse colour may reflect nonspecific binding or detection background rather than the reported cytoplasmic pattern (HPA: tissue IHC; general IHC practice).	Review blocking, washes, primary dilution and chromogen development against the validated IHC-P workflow; compare a no-primary section before scoring cell-level intensity (general IHC practice).
A brain cell population is negative despite reported brain expression.	UniProt reports brain expression, while HPA records cerebellar granular-layer cells as Not detected and caudate neuronal cells as Low by IHC (UniProt Q9UQN3 tissue specificity; HPA: tissue IHC).	Identify the sampled region and cell type before interpreting absence; do not equate tissue-level expression with a guaranteed IHC-positive cell population (HPA: tissue IHC; general IHC practice).
Q: Should an IF/ICC image show the same pattern?	HPA reports approved cytosolic ICC-IF localisation and rates antibody HPA052754 Supported for ICC; tissue IHC reliability remains Uncertain (HPA: subcellular ICC-IF; HPA: antibodies; HPA: tissue IHC reliability).	A: Use cytosolic localisation as a compartment check, while interpreting IF/ICC validation separately from chromogenic tissue IHC; follow the dedicated IF/ICC guide for its workflow (HPA: subcellular ICC-IF; HPA: antibodies).

Source: UniProt Q9UQN3 Human Protein Atlas tissue Human Protein Atlas subcellular

Experiment evidence

IHC & IF images showing CHMP2B staining

IHC analysis of CHMP2B using anti-CHMP2B antibody (M01935). CHMP2B was detected in a paraffin-embedded section of human colon cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1:50 rabbit anti-CHMP2B Antibody (M01935) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.

M01935 · IHCIHC analysis of CHMP2B using anti-CHMP2B antibody (M01935). CHMP2B was detected in a paraffin-embedded section of human colon cancer tissue.

IHC analysis of CHMP2B using anti-CHMP2B antibody (M01935). CHMP2B was detected in a paraffin-embedded section of human colon tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1:50 rabbit anti-CHMP2B Antibody (M01935) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.

M01935 · IHCIHC analysis of CHMP2B using anti-CHMP2B antibody (M01935). CHMP2B was detected in a paraffin-embedded section of human colon tissue. Heat m

HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000083937-CHMP2B/tissue/cerebral+cortex#imid_8456628

HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000083937-CHMP2B/tissue/colon#imid_8456554

HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000083937-CHMP2B/tissue/liver#imid_8456555

HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000083937-CHMP2B/tissue/kidney#imid_8456564

HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA052754; green = CHMP2B, blue = nucleus, red = microtubules.

← swipe / scroll for more images →

Real IHC / IF data behind the staining-pattern expectations in this section.

Sample controls for CHMP2B IHC & IF

🧪Run colon first: glandular cells should show CHMP2B staining (HPA: Medium in colon glandular cells). Use adipose tissue as the negative comparator: adipocytes are listed as not detected (HPA: Not detected in adipocytes); cells within the colon section that lack specific staining should show only background cytoplasmic signal, without assuming a particular cell type is negative.

Positive control tissue: Appendix (Glandular cells, HPA Medium)

Negative control tissue: Adipose tissue (HPA Not detected)

ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show CHMP2B in A-431, U-251MG, U2OS, NIH 3T3, with annotated localisation: Cytosol (approved) (HPA subcellular).

Technical controls: Include a no-primary (secondary-only) slide, a host- and clonality-matched rabbit isotype control, and a CHMP2B knockout specimen or validated peptide-block control (caption: rabbit primary; standard IHC practice). Block endogenous peroxidase and check colon sections for nonspecific DAB signal, especially in inflammatory cells (caption: HRP/DAB detection; standard IHC practice).

⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected M01935 caption does not state a fixative (selected-SKU caption: fixative not stated). The reported paraffin-section example used heat retrieval in EDTA at pH 8.0 and a 1:50 primary dilution overnight at 4°C; retrieval dependence has not been established by a comparison without retrieval (selected-SKU caption). Frozen sections have no supplied protocol, so an ease advantage over paraffin IHC is unreported; IF/ICC has HPA cytosolic images, but colon autofluorescence should be checked with a no-primary control (HPA: Cytosol approved; standard IF practice).

HPA tissue IHC evidence for CHMP2B

Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Uncertain — Low 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	Medium	Protein (IHC)	HPA →
Bone marrow	Hematopoietic cells	Medium	Protein (IHC)	HPA →
Bronchus	Respiratory epithelial cells	Medium	Protein (IHC)	HPA →
Colon	Glandular cells	Medium	Protein (IHC)	HPA →
Duodenum	Glandular cells	Medium	Protein (IHC)	HPA →

Undetected expression · recommended negative controls

Tissue	Cell type	Level	Evidence	Source
Adipose tissue	Adipocytes	Not detected	Protein (IHC)	HPA →
Adrenal gland	Glandular cells	Not detected	Protein (IHC)	HPA →
Breast	Adipocytes	Not detected	Protein (IHC)	HPA →
Cerebellum	Cells in granular layer	Not detected	Protein (IHC)	HPA →
Cerebral cortex	Endothelial cells	Not detected	Protein (IHC)	HPA →

Section 3

Advanced CHMP2B IHC Tips

Troubleshoot CHMP2B staining in paraffin sections by checking retrieval, cell identity, cytoplasmic localisation and assay controls before interpreting signal.

Which retrieval condition should I try first for CHMP2B in paraffin sections?

Start with heat-mediated retrieval in EDTA at pH 8.0 for paraffin sections (datasheet M01935). The selected image used that condition before 10% goat-serum blocking and a 1:50 primary incubation overnight at 4°C (caption M01935). If staining is weak, adjust heating time in matched sections while holding antibody dilution and detection conditions constant, then inspect tissue integrity and no-primary controls (standard IHC practice). Record both signal and background across the series; stronger DAB alone is unpersuasive if section edges, damaged areas or control slides also darken (standard IHC practice).

Could fixation explain weak or uneven CHMP2B staining?

The selected image documents a paraffin section but does not state its fixative, so CHMP2B-specific fixation sensitivity is unknown (caption M01935). Record the fixative, fixation duration and processing history for each specimen, and compare matched sections processed under documented conditions when staining varies (standard IHC practice). Fixation and processing can alter epitope accessibility in IHC generally, but the supplied evidence does not establish a CHMP2B-specific direction or threshold for that effect (standard IHC practice; caption M01935). Keep EDTA retrieval at pH 8.0 and the detection workflow consistent during that comparison, and assess morphology alongside signal (datasheet M01935; standard IHC practice).

Should CHMP2B appear diffuse or punctate in tissue sections?

CHMP2B is annotated in the cytosol and at the late-endosome membrane, with no transmembrane segment (UniProt Q9UQN3). Cytoplasmic staining can therefore be compatible with the annotation, while puncta may warrant closer inspection; neither pattern alone establishes the identity of the stained structure (UniProt Q9UQN3; standard IHC practice). In paraffin sections, compare staining within the intended cell population against adjacent cells, section edges and the no-primary control before calling discrete deposits positive (standard IHC practice). Treat predominantly nuclear staining cautiously because it does not match the supplied localisation annotations, and seek independent antibody or orthogonal support if it drives the conclusion (UniProt Q9UQN3; standard IHC practice).

Could isoforms or modified residues change what the antibody detects?

The record lists 2 CHMP2B isoforms, but the supplied antibody caption does not identify its epitope or establish isoform coverage (UniProt Q9UQN3; caption M01935). An N-terminal acetylation site at residue 2 and a reported phosphoserine at residue 199 are annotated; their effects on this antibody's staining are unknown (UniProt Q9UQN3). Request the immunogen or mapped epitope before assigning a negative section to a missing isoform or modification, and check whether that region is shared (standard IHC practice). If interpretation depends on isoform recognition, compare with an independently validated epitope or an orthogonal assay on matched material (standard IHC practice).

How should I cross-check CHMP2B localisation with multiplex IF?

Use IF as a localisation cross-check for the chromogenic IHC result, with a cell-identity marker such as an epithelial marker when evaluating colon glandular cells (HPA tissue IHC; standard IF practice). Choose spectrally separated fluorophores and favor a far-red CHMP2B channel if the tissue shows strong shorter-wavelength autofluorescence; include single-stain and no-primary controls (standard IF practice). Because CHMP2B is cytosolic and associates with the late-endosome membrane without a transmembrane segment, use permeabilisation appropriate for access to its cytosolic epitope (UniProt Q9UQN3; standard IF practice). The paraffin-section caption supplies no IF fixation or permeabilisation validation, so optimise those conditions within the separate IF workflow (caption M01935; standard IF practice).

What should I check when DAB background obscures CHMP2B?

The selected IHC example used 10% goat serum, a 1:50 primary overnight at 4°C, and a peroxidase-conjugated secondary for 30 minutes at 37°C (caption M01935). Check a no-primary section and apply an endogenous peroxidase block before DAB development to separate tissue enzyme activity from antibody-dependent signal (standard IHC practice). If background persists, assess blocking, washing, primary concentration and DAB development time one variable at a time in matched sections (standard IHC practice). Compare staining in intact cell bodies with deposits at section edges or damaged areas before accepting cytoplasmic signal as CHMP2B (UniProt Q9UQN3; standard IHC practice).

How can I quantify CHMP2B IHC without misleading cell-composition effects? ⚠ ANSWER MARKED FOR VERIFICATION

Define the cell population and cytoplasmic scoring compartment before reviewing outcomes, because CHMP2B is annotated in the cytosol and at the late-endosome membrane (UniProt Q9UQN3; standard IHC practice). For a defined population, report the percentage of positive cells and an H-score combining staining intensity and positive-cell fraction on a 0–300 scale (standard IHC practice). If counting discrete positive cells, report density per mm² of evaluable tissue and normalise comparisons to the same cell type or its total cell count (standard IHC practice). Keep thresholds, retrieval and imaging settings consistent, and report controls because the available tissue staining has uncertain reliability (HPA tissue IHC; standard IHC practice).

When is an apparent CHMP2B-positive section convincing?

A convincing result places antibody-dependent signal in the cytoplasm of morphologically intact cells, consistent with the cytosolic and late-endosome annotations (UniProt Q9UQN3; standard IHC practice). Check that the signal is absent from no-primary controls and does not track section edges, necrotic regions or endogenous peroxidase activity (standard IHC practice). Cell identity matters: the tissue profile reports medium staining in colon glandular cells and low staining in lung macrophages, yet rates the overall IHC evidence uncertain (HPA tissue IHC). Do not infer a universal positive or negative tissue from that profile; resolve unexpected nuclear or compartment-wide staining with independent antibody or orthogonal evidence (UniProt Q9UQN3; HPA tissue IHC; standard IHC practice).

Boster reagents

Best CHMP2B / Charged multivesicular body protein 2b IHC Antibodies

The catalog lists human, mouse and rat reactivity; M01935 has IHC images from human paraffin colon and colon cancer sections, while IF/ICC has no supplied figure (catalog: M01935).

Real IHC data

Anti-CHMP2B Rabbit Monoclonal Antibody

Cat # M01935

View product →

M01935 is the rendered card, with IHC images from human paraffin colon and colon cancer sections and listed IF/ICC applications (catalog: M01935 captions and applications). A01935-1 is also listed for IHC with human, mouse and rat reactivity, but has no supplied IHC image (catalog: A01935-1).

Which to pick: For tissue IHC, choose M01935 when the sample is a paraffin section: its own captions show human colon and colon cancer sections at 1:50 after EDTA pH 8 retrieval; the fixative is unreported (catalog: M01935 IHC captions). For IF/ICC, M01935 lists both applications and a 1:50 IF dilution, though no IF figure is supplied (catalog: M01935). Both SKUs list human, mouse and rat reactivity; M01935 is monoclonal and A01935-1 has no stated clone, but the supplied tissue images establish IHC results only in human samples (catalog: M01935 and A01935-1 reactivity; M01935 IHC captions).

Each figure is that product's own IHC / IF validation image from its datasheet.

References

UniProt Consortium. UniProt entry Q9UQN3 (CHM2B_HUMAN, Charged multivesicular body protein 2b).
Human Protein Atlas. CHMP2B tissue IHC expression (reliability: Uncertain).
Human Protein Atlas. CHMP2B subcellular location (ICC-IF): Localized to the cytosol..
Human Protein Atlas. CHMP2B antibody validation summary (2 antibodies).
Granulovacuolar degenerations appear in relation to hippocampal phosphorylated tau accumulation in various neurodegenerative disorders. PloS one 2011 — PMC3207829.
Molecular markers for granulovacuolar degeneration are present in rimmed vacuoles. PloS one 2013 — PMC3842945.
Early microgliosis precedes neuronal loss and behavioural impairment in mice with a frontotemporal dementia-causing CHMP2B mutation. Human molecular genetics 2017 — PMC5409096.
Identification of a PANoptosis-related Gene Signature for Predicting the Prognosis, Tumor Microenvironment and Therapy Response in Breast Cancer. Journal of Cancer 2024 — PMC10758028.
PubMed PMID:10810093 — UniProt-cited evidence.
PubMed PMID:11230166 — UniProt-cited evidence.
PubMed PMID:14702039 — UniProt-cited evidence.