MAP4K3 / Mitogen-activated protein kinase kinase kinase kinase 3 · IHC design guide

Design Immunohistochemistry for MAP4K3

Plan MAP4K3 IHC in paraffin sections using the catalog antibody at 2–5 μg/ml (datasheet A07188-2). Compare cytoplasmic staining across tissues (HPA tissue IHC) and include controls when interpreting variable staining (HPA tissue IHC: medium staining–RNA consistency).

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

Immunohistochemistry protocol sheet for MAP4K3 (IHC for MAP4K3): expected localisation Cytoplasmic staining in several tissues (HPA tissue IHC), antibody A07188-2, validated IHC image, and IHC protocol steps — Printable **MAP4K3** IHC protocol sheet — expected localisation Cytoplasmic staining in several tissues (HPA tissue IHC), antibody A07188-2, controls and protocol steps. Open the full MAP4K3 IHC guide →

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MAP4K3 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 several tissues (HPA tissue IHC)
	Staining pattern	High staining in fallopian tube ciliary rootlets (HPA tissue IHC)
	Antigen retrieval	EDTA pH 8.0 HIER, heat-mediated (datasheet A07188-2)
	Positive control ⓘ	Fallopian tube+4 more · see all
	Negative control ⓘ	Adipose tissue+4 more · see all

Important caveats

Reasons your staining may differ from the expected pattern.

	Fixation	Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A07188-2)
	Caveat	Staining and RNA show medium consistency (HPA tissue IHC)
	Regulation	Expression regulation unreported (UniProt)
	Isoform / epitope	3 isoforms; check epitope coverage (UniProt)

Section 1

Recommended MAP4K3 IHC & IF Protocols

The catalog antibody’s IHC-P protocol (datasheet A07188-2) is paired with one published MAP4K3 IHC protocol (PMC4467118).

Recommended immunohistochemistry (IHC-P) protocol parameters
Sample	Paraffin-embedded human skeletal muscle tissue; fixative not specified (datasheet A07188-2)
Fixation	Image fixative and duration unreported (datasheet A07188-2); 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 A07188-2); 20 min, 95–100 °C (standard)
Peroxidase block	3% H2O2, 10 min, room temperature (standard)
Blocking	10% goat serum (datasheet A07188-2)
Primary antibody	Rabbit anti-MAP4K3, 2-5 μg/ml (datasheet A07188-2)
Primary incubation	Overnight at 4 °C (datasheet A07188-2)
Detection	HRP-conjugated secondary, DAB chromogen (datasheet A07188-2)
Counterstain	Hematoxylin, blue, dehydrate and mount (standard)
Expected result	MAP4K3-positive staining in ciliated cells (ciliary rootlets) of fallopian tube (HPA tissue IHC: High). HPA tissue profile: Cytoplasmic expression in several tissues. No signal in the no-primary control.

💡Decision noteTry heat-mediated EDTA retrieval at pH 8.0 first (datasheet A07188-2); the published excerpt does not specify retrieval conditions (PMC4467118).

Section 2

What Is the Expected MAP4K3 Staining Pattern?

In paraffin section IHC, expect cytoplasmic MAP4K3 staining in selected cell populations, especially fallopian tube ciliated cell rootlets, with moderate staining reported in several other tissues (HPA: tissue IHC). MAP4K3 has no annotated transmembrane segment (UniProt Q8IVH8: topology). The tissue IHC profile is Approved, with medium consistency against RNA data and external verification pending (HPA: tissue IHC reliability).

What am I looking at on my slide?
Strong, localized staining at ciliary rootlets in fallopian tube ciliated cells.	This matches the highest reported tissue IHC signal and provides a useful positive reference. Assess staining in identified ciliated cells rather than treating the entire section as uniformly positive (HPA: High in fallopian tube ciliated cell rootlets).
Cytoplasmic staining in glomerular cells, lung macrophages, pancreatic exocrine cells, placental trophoblasts, or skeletal myocytes.	Moderate staining in these specified cell populations is compatible with the atlas pattern. Confirm cell identity from section morphology before scoring; an isolated stained structure should not define the whole tissue as positive (HPA: Medium in these cell populations).
Predominantly nuclear or sharply membranous staining without the expected cytoplasmic pattern.	Treat the compartment mismatch as a reason to investigate assay background or nonspecific binding. Tissue IHC reports cytoplasmic expression, while UniProt annotates no transmembrane segment; neither source alone proves that every unexpected signal is artefactual (HPA: cytoplasmic expression; UniProt Q8IVH8: topology).
Strong staining in cells reported as undetected, or widespread staining across unrelated cell types.	For example, adipocytes and adrenal glandular cells are reported as not detected. Check cell identification, primary antibody controls, and chromogen background before calling these cells MAP4K3 positive; an atlas negative is a reference observation, not proof of universal absence (HPA: adipocytes and adrenal glandular cells Not detected; standard IHC practice).
Diffuse color across tissue, including areas without a plausible cell pattern.	A diffuse result does not reproduce the reported cell and compartment distribution. Review the no primary control, blocking, washes, and chromogenic detection background before interpreting intensity as MAP4K3 expression (HPA: tissue IHC pattern; standard IHC practice).

💡Expected MAP4K3 appearanceCall positive when staining follows a cytoplasmic, cell identifiable pattern, with High signal at fallopian tube ciliated cell rootlets or Medium signal in listed positive populations; diffuse color or dominant staining in unrelated compartments warrants investigation (HPA: tissue IHC).

How each factor affects the staining
Cell population and tissue selection	Signal differs within and between tissues: fallopian tube ciliated cell rootlets are High, whereas lung macrophages and pancreatic exocrine cells are Medium. Select and score the named cells rather than assuming all cells in a positive tissue stain alike (HPA: tissue IHC).
Strength of the tissue IHC evidence	The profile is Approved but has medium consistency with RNA expression and awaits external verification. Use the listed pattern as an interpretation benchmark and investigate unexpected staining with assay controls (HPA: tissue IHC reliability; standard IHC practice).
Antibody validation	The listed rabbit polyclonal HPA030380 has IHC Approved status. The supplied record does not label its IHC result Enhanced, so agreement with an independently validated reagent cannot be assumed (HPA: antibody validation).
Protein topology and isoforms	MAP4K3 has no signal peptide or transmembrane segment and has 3 listed isoforms. These facts support an intracellular interpretation but do not identify which isoform an IHC antibody detects or establish an epitope location (UniProt Q8IVH8: topology and isoforms).
Chromogenic detection background	Endogenous activity relevant to the chosen detection system can produce color unrelated to the primary antibody. Interpret staining alongside a no primary control and apply an appropriate detection activity block when indicated (standard IHC practice).
IF/ICC Q&A: Where should MAP4K3 appear?	HPA reports a mainly centriolar satellite location in ICC-IF, with images from A-431, U-251MG, and U2OS. Use that as an IF localization reference; paraffin section IHC is described more broadly as cytoplasmic (HPA: subcellular ICC-IF and tissue IHC).

Why is my staining missing, weak or wrong?
Situation	Likely cause	Next action
No staining in fallopian tube ciliated cells.	The strongest listed positive reference is missing; assay failure or section quality may contribute, but the image alone cannot identify the cause (HPA: High in ciliated cell rootlets; standard IHC practice).	Check that ciliated cells are present and identifiable, then review primary antibody application, retrieval conditions, and detection controls against the validated IHC workflow (standard IHC practice).
Little or no staining in lung parenchyma despite an otherwise usable section.	The reported lung signal is in macrophages at Medium intensity, so a field lacking identifiable macrophages may appear negative (HPA: Medium in lung macrophages).	Inspect fields containing morphologically identifiable macrophages and compare with a named positive reference tissue; avoid scoring all lung cells as the expected positive population (HPA: lung macrophages; standard IHC practice).
Strong color appears in adipocytes or adrenal glandular cells.	Both populations are listed as Not detected, so the result conflicts with the atlas pattern; nonspecific signal or misidentified cells are possibilities (HPA: tissue IHC; standard IHC practice).	Recheck morphology and a no primary control, then review blocking and detection background before assigning MAP4K3 positivity (standard IHC practice).
Most cells show uniform, weak brown haze.	Diffuse chromogen can obscure the reported cell selective cytoplasmic pattern (HPA: tissue IHC; standard IHC practice).	Compare with the no primary control and review washing, blocking, and the chosen detection system. Score only cellular staining that remains interpretable above background (standard IHC practice).
Signal is predominantly nuclear or outlines plasma membranes.	The dominant compartment differs from the reported cytoplasmic IHC profile; no transmembrane segment is annotated (HPA: cytoplasmic expression; UniProt Q8IVH8: topology).	Check the positive reference and controls, then reassess antibody specificity and detection background before accepting the unexpected localization (standard IHC practice).
Weak staining in a cell population expected to be moderately positive.	HPA intensity categories describe observed patterns, and its Approved profile has medium RNA consistency with external verification pending; a single weak field is inconclusive (HPA: tissue IHC reliability and intensity).	Confirm cell identity, assess additional suitable fields, and compare the run with its positive and no primary controls before changing the interpretation (standard IHC practice).

Source: UniProt Q8IVH8 Human Protein Atlas tissue Human Protein Atlas subcellular

Experiment evidence

IHC & IF images showing MAP4K3 staining

IHC analysis of MAP4K3 using anti-MAP4K3 antibody (A07188-2). MAP4K3 was detected in a paraffin-embedded section of human skeletal muscle 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-MAP4K3 Antibody (A07188-2) 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.

A07188-2 · IHCIHC analysis of MAP4K3 using anti-MAP4K3 antibody (A07188-2). MAP4K3 was detected in a paraffin-embedded section of human skeletal muscle ti

HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000011566-MAP4K3/tissue/cerebral+cortex#imid_7702544

HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000011566-MAP4K3/tissue/colon#imid_7702478

HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000011566-MAP4K3/tissue/liver#imid_7702471

HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000011566-MAP4K3/tissue/kidney#imid_7702464

HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA030380; green = MAP4K3, blue = nucleus, red = microtubules.

← swipe / scroll for more images →

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

Sample controls for MAP4K3 IHC & IF

🧪Run fallopian tube first and look for staining in ciliated cell rootlets (HPA: High in fallopian tube ciliated cells). Use adipose tissue adipocytes as the negative comparison (HPA: Not detected in adipocytes); cells without ciliary rootlets on the positive slide should lack the rootlet staining pattern, though they need not be entirely unstained (HPA: ciliary rootlet localization).

Positive control tissue: Fallopian tube (Ciliated cells (ciliary rootlets), HPA High)

Negative control tissue: Adipose tissue (HPA Not detected)

ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show MAP4K3 in A-431, U-251MG, U2OS, with annotated localisation: Centriolar satellite (approved) (HPA subcellular).

Technical controls: Include a no-primary, secondary-only control and a concentration-matched rabbit IgG isotype control for the rabbit primary (caption: rabbit anti-MAP4K3; standard IHC practice); use a MAP4K3 knockout specimen or an immunizing-peptide block, if available, to test target specificity (standard IHC practice). For HRP–DAB detection, quench endogenous peroxidase in the fallopian tube section and check that any apical signal exceeds background in the no-primary control (caption: HRP–DAB; HPA: ciliary rootlets).

⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected A07188-2 paraffin-section caption does not state the fixative (caption: fixative not stated). Heat retrieval in EDTA at pH 8.0 is documented for a skeletal muscle section, but retrieval dependence in fallopian tube is unreported (caption: EDTA pH 8.0 in skeletal muscle). There is no supplied comparison showing that frozen sections or IF are easier; the small ciliary rootlet signal requires care when distinguishing staining from apical background (HPA: ciliary rootlet localization; standard IHC practice).

HPA tissue IHC evidence for MAP4K3

Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — Medium 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
Fallopian tube	Ciliated cells (ciliary rootlets)	High	Protein (IHC)	HPA →
Bronchus	Ciliated cells (ciliary rootlets)	Medium	Protein (IHC)	HPA →
Kidney	Cells in glomeruli	Medium	Protein (IHC)	HPA →
Lung	Macrophages	Medium	Protein (IHC)	HPA →
Nasopharynx	Ciliated cells (ciliary rootlets)	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 →
Appendix	Glandular cells	Not detected	Protein (IHC)	HPA →
Breast	Adipocytes	Not detected	Protein (IHC)	HPA →
Caudate	Glial cells	Not detected	Protein (IHC)	HPA →

Section 3

Advanced MAP4K3 IHC Tips

Troubleshoot MAP4K3 staining in paraffin sections by checking retrieval, cellular pattern, controls, and scoring before assigning biological meaning.

What retrieval should I try first for weak MAP4K3 staining in paraffin sections?

Start with heat-mediated antigen retrieval in EDTA at pH 8.0 for paraffin sections (datasheet A07188-2). The selected skeletal muscle example used this retrieval before overnight incubation at 4°C with 2 µg/ml antibody (datasheet A07188-2). If staining is weak, compare heating duration on adjacent sections while keeping antibody concentration, detection, and development constant (standard IHC practice). Include a skeletal muscle section with myocyte staining as a reference (HPA: Medium in skeletal muscle myocytes), and check whether stronger retrieval also increases diffuse background or damages tissue morphology (standard IHC practice).

Could fixation explain inconsistent MAP4K3 staining between paraffin blocks?

MAP4K3-specific sensitivity to fixation is unknown: the selected paraffin-section caption does not state a fixative (datasheet A07188-2). Compare blocks only after recording their fixative, fixation duration, processing history, section age, and retrieval conditions (standard IHC practice). For a controlled check, stain adjacent sections of approximately 4 µm thickness together and use the same EDTA pH 8.0 retrieval and antibody conditions (datasheet A07188-2; standard IHC practice). If signal differs, assess tissue preservation and internal staining patterns before attributing the difference to fixation, because the available evidence does not establish a MAP4K3-specific fixation effect.

Which MAP4K3 staining pattern is plausible in tissue sections?

Expect mainly cytoplasmic tissue staining, while judging the pattern within identifiable cell types (HPA: cytoplasmic expression in several tissues). The subcellular IF record places MAP4K3 mainly at centriolar satellites, but that fine structure may be unresolved in routine chromogenic sections (HPA: approved centriolar satellite location; standard microscopy practice). As tissue references, examine myocytes in skeletal muscle and exocrine glandular cells in pancreas, each reported at Medium staining (HPA: skeletal muscle myocytes; HPA: pancreatic exocrine cells). Treat isolated strong nuclear or membranous staining cautiously, since UniProt gives no subcellular annotation and records no transmembrane segment (UniProt Q8IVH8 topology and subcellular record).

How should isoforms and epitope uncertainty affect MAP4K3 IHC interpretation?

MAP4K3 has 3 annotated isoforms, so an antibody result cannot establish which isoform is present unless its epitope and isoform coverage are known (UniProt Q8IVH8 isoforms; standard antibody interpretation). Check the catalog antibody’s immunogen or epitope documentation before comparing staining across tissues, and report that information if available (standard IHC practice). The protein contains a kinase domain at residues 16–273 and a CNH domain at 556–867, with annotated phosphorylation at Ser329 and Ser398 (UniProt Q8IVH8). Those features alone do not show whether fixation or retrieval exposes this antibody’s epitope; resolve unexpected patterns with an independently validated reagent or orthogonal evidence (standard IHC practice).

How can IF help assess MAP4K3 staining in the expected cells?

On the separate IF/ICC workflow, multiplex MAP4K3 with a marker that identifies the cell population being examined, such as a validated myocyte marker in skeletal muscle (HPA: Medium in skeletal muscle myocytes; standard IF practice). Choose spectrally separated fluorophores and place a relatively bright far-red signal on the weaker target when tissue autofluorescence obscures shorter wavelengths (standard IF practice). MAP4K3 has no annotated transmembrane segment, and IF images support a centriolar satellite location, so evaluate permeabilisation for access to intracellular epitopes (UniProt Q8IVH8 topology; HPA: approved centriolar satellite location). Include single-label and secondary-only controls to distinguish colocalisation from bleed-through and background (standard IF practice).

What should I change when MAP4K3 DAB staining is diffuse?

First inspect the no-primary control for secondary reagent binding and endogenous peroxidase signal, then check whether DAB development has obscured cellular boundaries (standard chromogenic IHC practice). The selected protocol blocked tissue with 10% goat serum and used a peroxidase-conjugated anti-rabbit secondary for 30 minutes at 37°C (datasheet A07188-2). Keep the EDTA pH 8.0 retrieval fixed while adjusting antibody concentration or development time one variable at a time (datasheet A07188-2; standard IHC practice). Compare the result with cell-associated cytoplasmic staining, rather than accepting uniform pigment across tissue, because the HPA tissue profile describes cytoplasmic expression (HPA: tissue IHC profile).

How should I score MAP4K3 IHC across samples? ⚠ ANSWER MARKED FOR VERIFICATION

Define the anatomical compartment and eligible cell type before scoring, because reported staining varies by cell population within tissues (HPA: tissue IHC profile). For cytoplasmic DAB, record the percentage of positive cells and intensity on a 0–3 scale, then calculate an H-score from 0–300 if intensity matters (standard IHC scoring practice). Alternatively, report positive-cell density per mm² when cell abundance is the biological endpoint (standard IHC scoring practice). Normalise to the number of eligible cells or viable tissue area, use matched imaging and DAB thresholds, and exclude folds, edges, and necrotic regions by a predefined rule (standard quantitative IHC practice).

How can I distinguish convincing MAP4K3 staining from artefact?

Look for reproducible cell-associated cytoplasmic staining in expected populations, such as skeletal muscle myocytes or pancreatic exocrine cells, both reported at Medium levels (HPA: skeletal muscle myocytes; HPA: pancreatic exocrine cells). Question signal confined to section edges, folds, necrosis, or all cell types equally, and compare it with no-primary and peroxidase-block controls (standard chromogenic IHC practice). Strong exclusively membranous staining needs independent support because MAP4K3 has no annotated transmembrane segment (UniProt Q8IVH8 topology). HPA rates its tissue antibody evidence Approved with medium consistency against RNA and pending external verification, so use orthogonal evidence before making a definitive biological claim (HPA: tissue IHC reliability).

Boster reagents

Best MAP4K3 / Mitogen-activated protein kinase kinase kinase kinase 3 IHC Antibodies

The IHC-validated anti-MAP4K3 antibody has an image from a human skeletal muscle paraffin section (catalog IHC caption); its listed species reactivity is human, mouse and rat (catalog reactivity).

Real IHC data

Anti-MAP4K3 Antibody ®

Cat # A07188-2

View product →

A07188-2 will render with an IHC image from a human skeletal muscle paraffin section (catalog IHC caption). Its application list includes IHC, and its listed reactivity covers human, mouse and rat (catalog applications and reactivity).

Which to pick: For tissue IHC, choose A07188-2: its caption documents a human skeletal muscle paraffin section stained at 2 μg/ml after heat retrieval in EDTA, pH 8.0 (catalog IHC caption). The caption does not report the fixative (catalog IHC caption). A07188-2 has no listed IF/ICC application or IF image, so it is not an evidenced IF/ICC choice; for cross-species work, it lists mouse and rat reactivity, while the supplied IHC image documents human tissue only (catalog applications, reactivity and IHC caption).

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

References

UniProt Consortium. UniProt entry Q8IVH8 (M4K3_HUMAN, Mitogen-activated protein kinase kinase kinase kinase 3).
Human Protein Atlas. MAP4K3 tissue IHC expression (reliability: Approved).
Human Protein Atlas. MAP4K3 subcellular location (ICC-IF): Mainly localized to the centriolar satellites..
Human Protein Atlas. MAP4K3 antibody validation summary (1 antibodies).
Inhibition of EP2/EP4 signaling abrogates IGF-1R-mediated cancer cell growth: involvement of protein kinase C-θ activation. Oncotarget 2015 — PMC4467118.
PubMed PMID:9275185 — UniProt-cited evidence.
PubMed PMID:15489334 — UniProt-cited evidence.
PubMed PMID:19369195 — UniProt-cited evidence.