TRPM4 / Transient receptor potential cation channel subfamily M member 4 · IHC design guide

Design Immunohistochemistry for TRPM4

Plan chromogenic IHC on paraffin sections using gastrointestinal or prostate glandular cells as positive controls (HPA tissue IHC). Compare cytoplasmic and membranous staining with the tissue profile, starting the catalog antibody at 2–5 μg/mL (HPA tissue IHC; datasheet PB9902).

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

Immunohistochemistry protocol sheet for TRPM4 (IHC for TRPM4): expected localisation Cytoplasmic and membranous tissue staining (HPA tissue IHC), antibody PB9902, validated IHC image, and IHC protocol steps — Printable **TRPM4** IHC protocol sheet — expected localisation Cytoplasmic and membranous tissue staining (HPA tissue IHC), antibody PB9902, controls and protocol steps. Open the full TRPM4 IHC guide →

Navigate to: Protocols Staining pattern Controls Antibodies

TRPM4 Immunohistochemistry Experimental Design Guide

Expected localisation, validated protocols, controls and antibodies — the at-a-glance facts below, then the full design guide.

Must know before staining

	Expected localisation	Cytoplasmic and membranous tissue staining (HPA tissue IHC)
	Staining pattern	Glandular cells show cytoplasmic and membranous staining (HPA tissue IHC)
	Antigen retrieval	EDTA pH 8.0 HIER, heat-mediated (datasheet PB9902)
	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 formalin fixation consistent across sections. (standard IHC practice; not target-specific)
	Caveat	Antibody staining has medium consistency with RNA data (HPA tissue IHC)
	Regulation	Higher expression in Th2 than Th1 cells (UniProt)
	Isoform / epitope	3 isoforms; check whether the epitope is cytoplasmic or extracellular (UniProt)

Section 1

Recommended TRPM4 IHC & IF Protocols

The catalog antibody’s IHC-P protocol is accompanied by published TRPM4 staining protocols for cerebral infarcts, rat organs, and prostate cancer samples (PMC4620032; PMC4383534; PMC13290428).

Recommended immunohistochemistry (IHC-P) protocol parameters
Sample	Paraffin-embedded human prostate cancer tissue; fixative not specified (datasheet PB9902)
Fixation	Image fixative and duration unreported (datasheet PB9902); 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 PB9902); 20 min, 95–100 °C (standard)
Peroxidase block	3% H2O2, 10 min, room temperature (standard)
Blocking	10% goat serum (datasheet PB9902)
Primary antibody	Rabbit anti-TRPM4, 2-5μg/ml (datasheet PB9902)
Primary incubation	Overnight at 4 °C (datasheet PB9902)
Detection	HRP-conjugated secondary, DAB chromogen (datasheet PB9902)
Counterstain	Hematoxylin, blue, dehydrate and mount (standard)
Expected result	TRPM4-positive staining in glandular cells of appendix (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic and membranous expression in several different tissue types, most abundant in gastrointestinal tract and prostate. No signal in the no-primary control.

💡Decision noteStart with heat-mediated EDTA, pH 8.0, for the catalog antibody (PB9902 datasheet); optimize retrieval for other antibodies and samples.

Section 2

What Is the Expected TRPM4 Staining Pattern?

In paraffin-section IHC, expect TRPM4 staining along cell borders and in cytoplasm, especially in gastrointestinal and prostate glandular cells (HPA tissue IHC: high; reliability Enhanced, with medium staining–RNA consistency). Membrane staining accords with six transmembrane segments; cytoplasmic staining is compatible with endoplasmic reticulum and Golgi localisation (UniProt Q8TD43 topology and subcellular location).

What am I looking at on my slide?
Glandular cells show distinct membranous and cytoplasmic staining in colon, small intestine, stomach or prostate.	This fits the reported high staining in those cell types and the broader cytoplasmic and membranous tissue profile (HPA tissue IHC). Compare the stained cells with gland morphology before scoring intensity; a dark region alone does not establish which cells are positive (general IHC interpretation).
A section shows predominantly nuclear staining, with little staining at cell borders or in cytoplasm.	That is discordant with the reported tissue IHC profile (HPA tissue IHC: cytoplasmic and membranous). Check localisation and controls before assigning it to TRPM4 (general IHC practice). Nucleoplasmic localisation is reported separately for ICC-IF and should not be treated as the expected paraffin-section pattern (HPA subcellular ICC-IF).
Strong colour appears mainly in adipocytes or lymph-node germinal centre cells.	These cell types are reported as not detected in the sampled tissue IHC observations (HPA tissue IHC). Unexpected staining warrants checks for antibody cross-reactivity or endogenous detection activity (general IHC practice). A not-detected HPA observation is a comparison point, not proof that every specimen must be negative.
Colour spreads evenly over tissue, stroma and empty areas rather than following cell borders or cytoplasm.	The distribution does not resemble the reported cellular pattern (HPA tissue IHC). Review background in the negative reagent control, blocking and chromogen development (general IHC practice); background alone cannot identify whether antibody binding or detection chemistry caused the colour.
No staining appears in a prostate or gastrointestinal glandular section selected as a positive control.	Those glandular cells have high reported staining, so absence of signal calls for a technical review (HPA tissue IHC). Confirm tissue identity, control performance, reagent delivery and detection before interpreting a test section as negative (general IHC practice); expression can vary among specimens.

💡Expected TRPM4 appearanceA convincing positive shows clear cytoplasmic and membranous staining in gastrointestinal or prostate glandular cells, where HPA reports high staining, while isolated nuclear colour or uniform background is a suspect IHC result (HPA tissue IHC; general IHC interpretation).

How each factor affects the staining
Which locations guide scoring?	Six membrane-spanning segments and annotated cell-membrane, endoplasmic-reticulum and Golgi localisation support assessing both border and cytoplasmic colour (UniProt Q8TD43 topology and subcellular location). HPA's tissue IHC profile reports both compartments; it does not specify an expected border-to-cytoplasm intensity ratio (HPA tissue IHC).
How strong is the tissue reference?	The tissue profile is labelled Enhanced, with medium consistency between staining and RNA expression (HPA tissue IHC). The listed rabbit polyclonal antibody HPA041169 is IHC Enhanced (HPA antibodies); that validation status supports use of the pattern as a reference without guaranteeing identical staining in every specimen.
Do sequence features dictate retrieval?	TRPM4 has three annotated isoforms and one glycosylation site at residue 992 (UniProt Q8TD43). These annotations do not identify the antibody epitope or establish an antigen-retrieval condition. Use the IHC-validated antibody's documented procedure and optimise retrieval with controls if needed (general IHC practice).
IF/ICC Q&A: should nuclei stain?	HPA reports approved nucleoplasmic and supported plasma-membrane localisation in ICC-IF (HPA subcellular ICC-IF). That observation helps interpret IF/ICC images but does not override the cytoplasmic and membranous paraffin-section IHC pattern (HPA tissue IHC). Follow the separate IF/ICC guide for that application.

Why is my staining missing, weak or wrong?
Situation	Likely cause	Next action
Positive-control glandular cells are blank.	A failed staining or detection step is possible; high staining is reported in gastrointestinal and prostate glandular cells (HPA tissue IHC).	Check the IHC-validated antibody procedure, reagent application, retrieval and detection with a concurrent positive control (general IHC practice). Do not score the test section as negative until the control works.
Every structure is dark, including spaces outside cells.	Widespread colour is inconsistent with HPA's cellular cytoplasmic and membranous profile (HPA tissue IHC); excess background may arise during antibody or detection steps (general IHC practice).	Compare negative reagent controls; review blocking, washes, antibody concentration and chromogen development, changing one condition at a time (general IHC practice).
Staining is confined to nuclei in paraffin sections.	Nuclear-only staining differs from the reported tissue IHC distribution (HPA tissue IHC), although nucleoplasm is reported in ICC-IF (HPA subcellular ICC-IF).	Check the positive tissue control and antibody specificity, then score IHC localisation separately from ICC-IF observations (general IHC interpretation).
Adipocytes or germinal centre cells stain prominently.	HPA reports those sampled cell types as not detected (HPA tissue IHC); cross-reactivity or endogenous detection activity is possible (general IHC practice).	Inspect a reagent negative control and confirm cell identity and staining distribution; repeat with validated controls before treating unexpected colour as TRPM4 (general IHC practice).
Staining appears in some glands but not adjacent glands.	A mixed result cannot be resolved from a tissue-level reference alone; HPA reports high glandular staining in several tissues but does not specify uniformity within each section (HPA tissue IHC).	Assess morphology and preservation, compare control and test sections stained together, and record the fraction of positive glands as well as intensity (general IHC practice).
An IF/ICC image and an IHC section give different nuclear patterns.	The reported application profiles differ: nucleoplasm is approved in ICC-IF, while tissue IHC is cytoplasmic and membranous (HPA subcellular ICC-IF; HPA tissue IHC).	Interpret each image against its own application-specific reference and controls; investigate an IHC nuclear-only result as an unexpected IHC pattern (general IHC interpretation).

Source: UniProt Q8TD43 Human Protein Atlas tissue Human Protein Atlas subcellular

Experiment evidence

IHC & IF images showing TRPM4 staining

IHC analysis of TRPM4 using anti-TRPM4 antibody (PB9902). TRPM4 was detected in a paraffin-embedded section of human prostate 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 2 μg/ml rabbit anti-TRPM4 Antibody (PB9902) 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.

PB9902 · IHCIHC analysis of TRPM4 using anti-TRPM4 antibody (PB9902). TRPM4 was detected in a paraffin-embedded section of human prostate cancer tissue.

IHC analysis of TRPM4 using anti-TRPM4 antibody (PB9902). TRPM4 was detected in a paraffin-embedded section of human squamous cell carcinoma of cervix 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 2 μg/ml rabbit anti-TRPM4 Antibody (PB9902) 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.

PB9902 · IHCIHC analysis of TRPM4 using anti-TRPM4 antibody (PB9902). TRPM4 was detected in a paraffin-embedded section of human squamous cell carcinoma

HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000130529-TRPM4/tissue/cerebral+cortex#imid_10973755

HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000130529-TRPM4/tissue/colon#imid_10973681

HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000130529-TRPM4/tissue/liver#imid_10973690

HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000130529-TRPM4/tissue/kidney#imid_10973691

HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA041169; green = TRPM4, blue = nucleus, red = microtubules.

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

Sample controls for TRPM4 IHC & IF

🧪Run prostate first: glandular cells should stain (HPA: High in prostate glandular cells). Use epididymis glandular cells as a tissue negative (HPA: Not detected in epididymis glandular cells); on the prostate slide, compare surrounding nonglandular cells with the no-primary control for background-level staining rather than assuming they are TRPM4-negative.

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

Negative control tissue: Adipose tissue (HPA Not detected)

ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show TRPM4 in A-431, U-251MG, U2OS, with annotated localisation: Nucleoplasm (approved), Plasma membrane (supported) (HPA subcellular).

Technical controls: Include no-primary (secondary-only) and concentration-matched rabbit IgG isotype controls alongside a TRPM4 knockout specimen, if available, as a biological negative (selected PB9902 caption: rabbit primary antibody). For chromogenic detection, quench endogenous peroxidase and check the prostate section for residual background before interpreting glandular DAB signal (selected PB9902 caption: HRP/DAB detection).

⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the exact PB9902 tissue-IHC caption does not state the fixative. The caption uses heat retrieval in EDTA at pH 8.0 on a paraffin prostate cancer section, but does not establish that retrieval is required for all TRPM4 specimens (selected PB9902 caption). The supplied evidence does not establish whether frozen sections or IF are easier, or identify a prostate-specific artefact; assess nonspecific glandular background with the controls above.

HPA tissue IHC evidence for TRPM4

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
Appendix	Glandular cells	High	Protein (IHC)	HPA →
Colon	Glandular cells	High	Protein (IHC)	HPA →
Duodenum	Glandular cells	High	Protein (IHC)	HPA →
Prostate	Glandular cells	High	Protein (IHC)	HPA →
Rectum	Glandular cells	High	Protein (IHC)	HPA →

Undetected expression · recommended negative controls

Tissue	Cell type	Level	Evidence	Source
Adipose tissue	Adipocytes	Not detected	Protein (IHC)	HPA →
Caudate	Glial cells	Not detected	Protein (IHC)	HPA →
Cerebellum	Cells in granular layer	Not detected	Protein (IHC)	HPA →
Epididymis	Glandular cells	Not detected	Protein (IHC)	HPA →
Hippocampus	Glial cells	Not detected	Protein (IHC)	HPA →

Section 3

Advanced TRPM4 IHC Tips

TRPM4 staining should be assessed by cell type and compartment, with the paraffin section workflow anchored to the catalog antibody’s documented retrieval and detection conditions.

What retrieval should I start with for TRPM4 in paraffin sections?

Start with heat mediated antigen retrieval in EDTA at pH 8.0 for paraffin sections stained with PB9902 (datasheet PB9902). The documented prostate cancer section then received 2 μg/ml primary antibody overnight at 4°C, so keep those conditions consistent while comparing retrieval runs (caption PB9902). If staining is weak, compare a shorter and longer heating interval on adjacent sections, keeping cooling and wash steps consistent across slides (standard IHC practice). Judge retrieval by glandular cell staining and tissue integrity together, because the expected tissue pattern includes cytoplasmic and membranous signal in prostate glands (HPA tissue IHC).

How should I troubleshoot fixation when TRPM4 staining is weak?

Target specific fixation sensitivity is unknown: the PB9902 tissue caption identifies a paraffin section but does not state its fixative (caption PB9902). Record the fixative, fixation duration, and processing history before comparing specimens, since these variables can change antigen accessibility in paraffin IHC (standard IHC practice). On matched sections, hold EDTA retrieval at pH 8.0 and primary antibody at 2 μg/ml constant while investigating processing differences (datasheet PB9902; caption PB9902). Include a similarly processed prostate glandular control in each run, and interpret a weak case only after that control stains appropriately (HPA tissue IHC; standard IHC practice).

Which compartments should contain credible TRPM4 staining?

Expect staining at the cell membrane, with possible intracellular signal associated with the endoplasmic reticulum and Golgi apparatus (UniProt Q8TD43 localisation). Cytoplasmic and membranous staining across several tissues, especially gastrointestinal tract and prostate, is also reported in tissue IHC (HPA tissue IHC). Because TRPM4 has 6 transmembrane segments and long cytoplasmic regions, score membrane and cytoplasmic patterns separately rather than treating diffuse colour as one result (UniProt Q8TD43 topology; standard IHC practice). A predominantly nuclear result needs independent assessment: nucleoplasm is reported in cell based imaging, but the cited tissue IHC profile emphasises cytoplasmic and membranous staining (HPA subcellular; HPA tissue IHC).

Can isoforms or epitope position change my IHC interpretation?

TRPM4 has 3 annotated isoforms, but the supplied PB9902 caption does not map its epitope or establish isoform coverage (UniProt Q8TD43 isoforms; caption PB9902). Its 6 membrane spanning segments separate cytoplasmic regions from short extracellular regions, making epitope position relevant to accessibility after processing (UniProt Q8TD43 topology; standard IHC practice). The annotated glycosylation site at residue 992 and phosphoserines at 1145 and 1152 do not establish an effect on this antibody without epitope information (UniProt Q8TD43 modifications; caption PB9902). Check an available immunogen or epitope map before claiming isoform specificity, and report any unresolved coverage as unknown (standard IHC practice).

How should I adapt TRPM4 assessment for multiplex IF?

For multiplex IF, pair TRPM4 with a validated glandular cell marker when assessing prostate sections, where glandular cells show high tissue IHC staining (HPA tissue IHC; standard IF practice). Choose a far red TRPM4 fluorophore when the specimen shows strong shorter wavelength autofluorescence, and inspect unstained and single stained controls before assigning overlap (standard IF practice). Select permeabilisation only after checking epitope position: cytoplasmic TRPM4 regions require intracellular access, whereas an accessible extracellular epitope may be assessed without detergent (UniProt Q8TD43 topology; standard IF practice). The PB9902 paraffin IHC conditions do not establish an IF dilution or fixation condition; optimise those variables for the IF assay (caption PB9902; standard IF practice).

What should I check when TRPM4 DAB staining is widespread?

The documented PB9902 paraffin workflow used 10% goat serum blocking, a peroxidase conjugated secondary antibody, and DAB detection (caption PB9902). If colour appears in unlikely cells or throughout the section, inspect a no primary control and add an endogenous peroxidase block as needed (standard IHC practice). Titrate around the documented 2 μg/ml primary concentration while keeping EDTA retrieval at pH 8.0 fixed, then compare staining in matched sections (caption PB9902; datasheet PB9902; standard IHC practice). Record edge and necrotic staining separately from viable glandular cells, since the expected tissue profile is cell associated and cytoplasmic or membranous (HPA tissue IHC; standard IHC practice).

How should I score TRPM4 staining across sections? ⚠ ANSWER MARKED FOR VERIFICATION

Define the scored cell population before analysis, such as prostate glandular cells, and record membranous and cytoplasmic signal separately (HPA tissue IHC; standard IHC practice). For each compartment, report percentage positive and an H score using intensity grades 0–3, yielding a possible score of 0–300 (standard IHC practice). Normalise positive cell counts to the number of viable cells assessed, or staining density to viable tissue area in mm² (standard IHC practice). Apply the same thresholds and imaging conditions across batches, with a prostate glandular control to monitor run variation (HPA tissue IHC; standard IHC practice).

How can I distinguish true TRPM4 signal from artefact?

A convincing result follows cellular boundaries or cytoplasm in the expected cell population and appears consistently across intact regions of the section (HPA tissue IHC; standard IHC practice). Prostate glandular cells offer a high staining reference, whereas adipocytes were reported as not detected in the cited tissue survey (HPA tissue IHC). Treat staining limited to section edges, necrotic material, or a no primary control as suspect, and check endogenous peroxidase activity in the DAB workflow (standard IHC practice). Assess nucleus only staining cautiously because cell based imaging reports nucleoplasm, while the tissue IHC profile mainly reports cytoplasmic and membranous expression with medium RNA concordance (HPA subcellular; HPA tissue IHC).

Boster reagents

Best TRPM4 / Transient receptor potential cation channel subfamily M member 4 IHC Antibodies

PB9902 has real IHC data from human paraffin-embedded prostate cancer and cervical squamous cell carcinoma sections (catalog: PB9902 IHC captions). No IF data are supplied (catalog: PB9902 IF images).

Real IHC data

Anti-TRPM4 Antibody ®

Cat # PB9902

View product →

PB9902 is the only SKU shown and is listed for human IHC (catalog: PB9902 applications and reactivity). Its IHC captions document staining in paraffin-embedded human prostate cancer and cervical squamous cell carcinoma sections (catalog: PB9902 IHC captions).

Which to pick: Choose PB9902 for human tissue IHC: its own figure documents a paraffin-embedded prostate cancer section (PB9902 IHC figure caption). No IF/ICC recommendation is supported because IF/ICC is unlisted and no IF image is supplied (catalog: PB9902 applications and IF images). Cross-species use is unsupported because reactivity is listed only for Human; the IHC caption does not report the fixative (catalog: PB9902 reactivity; PB9902 IHC figure caption).

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

References

UniProt Consortium. UniProt entry Q8TD43 (TRPM4_HUMAN, Transient receptor potential cation channel subfamily M member 4).
Human Protein Atlas. TRPM4 tissue IHC expression (reliability: Enhanced).
Human Protein Atlas. TRPM4 subcellular location (ICC-IF): Localized to the nucleoplasm and plasma membrane..
Human Protein Atlas. TRPM4 antibody validation summary (1 antibodies).
Water channel changes in astrocytes associated with buprenorphine administration in a rat model of diffuse traumatic brain injury. Research square 2026 — PMC13060523.
Sur1-Trpm4 Cation Channel Expression in Human Cerebral Infarcts. Journal of neuropathology and experimental neurology 2015 — PMC4620032.
Transient receptor potential melastatin-4 is involved in hypoxia-reoxygenation injury in the cardiomyocytes. PloS one 2015 — PMC4383534.
TRPM4 Expression as a Predictive Biomarker and a Mechanistic Driver of Acetalax Activity in Prostate Cancer: Preclinical Efficacy Studies. Molecular cancer therapeutics 2026 — PMC13290428.
PubMed PMID:11535825 — UniProt-cited evidence.
PubMed PMID:12015988 — UniProt-cited evidence.
PubMed PMID:12842017 — UniProt-cited evidence.