CAMLG / Guided entry of tail-anchored proteins factor CAMLG · IHC design guide

Design Immunohistochemistry for CAMLG

Plan CAMLG staining in paraffin sections using the IHC-validated antibody at 2–5 μg/ml (datasheet A08322-2). Use the observed cytoplasmic tissue pattern to interpret staining (HPA tissue IHC), alongside CAMLG’s ER membrane location (UniProt).

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

Immunohistochemistry protocol sheet for CAMLG (IHC for CAMLG): expected localisation Cytoplasmic tissue staining (HPA tissue IHC); ER membrane (UniProt), antibody A08322-2, validated IHC image, and IHC protocol steps — Printable **CAMLG** IHC protocol sheet — expected localisation Cytoplasmic tissue staining (HPA tissue IHC); ER membrane (UniProt), antibody A08322-2, controls and protocol steps. Open the full CAMLG IHC guide →

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CAMLG 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 tissue staining (HPA tissue IHC); ER membrane (UniProt)
	Staining pattern	Cytoplasmic in cerebral neurons and fallopian glandular cells (HPA tissue IHC)
	Antigen retrieval	EDTA pH 8.0 HIER, heat-mediated (datasheet A08322-2)
	Positive control ⓘ	Cerebral cortex+4 more · see all
	Negative control ⓘ	Adipose tissue+3 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	Staining and RNA expression show medium consistency (HPA tissue IHC)
	Regulation	Highest in brain, testis and ovary (UniProt)
	Isoform / epitope	0 isoforms annotated; epitope side may affect access (UniProt)

Section 1

Recommended CAMLG IHC & IF Protocols

Compare the catalog antibody’s IHC-P protocol (datasheet A08322-2) with one published FFPE brain protocol (PMC12093335).

Recommended immunohistochemistry (IHC-P) protocol parameters
Sample	Paraffin-embedded human breast cancer tissue; fixative not specified (datasheet A08322-2)
Fixation	Image fixative and duration unreported (datasheet A08322-2); 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 A08322-2); 20 min, 95–100 °C (standard)
Peroxidase block	3% H2O2, 10 min, room temperature (standard)
Blocking	10% goat serum (datasheet A08322-2)
Primary antibody	Rabbit anti-CAMLG, 2-5 μg/ml (datasheet A08322-2)
Primary incubation	Overnight at 4 °C (datasheet A08322-2)
Detection	HRP-conjugated secondary, DAB chromogen (datasheet A08322-2)
Counterstain	Hematoxylin, blue, dehydrate and mount (standard)
Expected result	CAMLG-positive staining in neuronal cells of cerebral cortex (HPA tissue IHC: High). HPA tissue profile: General cytoplasmic expression. No signal in the no-primary control.

💡Decision noteStart with heat-mediated EDTA pH 8.0 retrieval (datasheet A08322-2); the published protocol used citric acid at 95°C for 30 min (PMC12093335).

Section 2

What Is the Expected CAMLG Staining Pattern?

CAMLG is an endoplasmic reticulum (ER) membrane protein with three transmembrane segments and cytoplasmic and lumenal regions (UniProt P49069 topology). In paraffin-section IHC, expect a mainly cytoplasmic pattern, strongest in cerebral cortex neuronal cells and fallopian tube glandular cells (HPA: High; general cytoplasmic expression). HPA rates the tissue IHC profile Approved, with medium consistency between antibody staining and RNA expression (HPA: tissue IHC reliability).

What am I looking at on my slide?
Clear cytoplasmic staining in cerebral cortex neuronal cells or fallopian tube glandular cells.	This matches the reported high-staining cell populations (HPA: High in both). Judge the stained cells, rather than assigning one intensity to the whole tissue; the expected compartment is cytoplasmic in IHC (HPA: general cytoplasmic expression).
Predominantly nuclear or cell-surface staining, with little cytoplasmic signal.	Treat this as a compartment mismatch needing investigation, not proof of CAMLG localisation (HPA: general cytoplasmic IHC expression; UniProt P49069: ER membrane). Check whether the pattern persists in a known-positive section and with an appropriate negative control (general IHC practice).
Strong staining of adipocytes, skeletal myocytes, or soft-tissue fibroblasts.	These cell types were reported as not detected in the supplied tissue IHC profile (HPA: adipose tissue, skeletal muscle, soft tissue). Consider antibody cross-reactivity or endogenous detection activity, particularly if nearby expected-positive cells do not show the reported cytoplasmic pattern (general IHC practice).
Diffuse colour across cells and surrounding tissue, obscuring cell boundaries.	A broad haze cannot establish the reported cell-specific cytoplasmic pattern (HPA: tissue IHC profile). Check a section processed without primary antibody for detection-system background, then review blocking, washes and detection exposure (general IHC practice).
No convincing signal in cerebral cortex neuronal cells or fallopian tube glandular cells.	Both are reported high-staining populations, so an absent result warrants an assay check (HPA: High in both). Examine section quality and controls, then review antigen retrieval, antibody conditions and detection steps as general IHC variables; the supplied sources do not establish CAMLG-specific retrieval or fixation sensitivity.

💡Expected CAMLG appearanceCall a positive IHC result when identifiable neuronal or glandular cells show clear, mainly cytoplasmic staining, potentially high in cerebral cortex or fallopian tube (HPA: general cytoplasmic expression; High in those cells); dominant nuclear-only staining or strong signal in reported not-detected cells is a warning sign (HPA: tissue IHC profile).

How each factor affects the staining
Cell population and tissue	High staining is reported in cerebral cortex neuronal cells and fallopian tube glandular cells; medium staining occurs in several other listed cell populations (HPA: tissue IHC profile). Low or not-detected categories describe those sampled cells and should guide control choice without making a whole tissue an absolute negative.
Compartment and antibody epitope	CAMLG has three membrane-spanning segments, with residues 1–189 and 232–269 cytoplasmic and two short lumenal regions (UniProt P49069 topology). Epitope location could affect access in an assay, but no antibody epitope or target-specific retrieval requirement is supplied; do not infer one from topology.
IHC evidence strength	HPA calls the tissue IHC profile Approved and reports medium consistency with RNA expression (HPA: tissue IHC reliability). The listed antibodies HPA052636 and HPA056472 are each IHC Approved (HPA: antibody validation); this supports using the pattern as a reference, while leaving discrepant slides open to control-based review.
Does IF/ICC confirm the IHC compartment?	Not decisively: HPA reports mainly vesicular localisation, with additional nucleoplasm and nucleoli, and marks these locations uncertain (HPA: subcellular ICC-IF). HPA052636 is ICC Uncertain, while no ICC status is supplied for HPA056472 (HPA: antibody validation). Use the tissue IHC profile to interpret paraffin sections.
Chromogenic detection background	Endogenous detection activity and nonspecific colour can obscure cell-level localisation in chromogenic IHC (general IHC practice). A no-primary control helps identify detection-system signal; it does not establish whether a remaining primary-dependent signal is specific (general IHC practice).

Why is my staining missing, weak or wrong?
Situation	Likely cause	Next action
Known-positive cells are blank.	An assay step or section issue may have reduced detectable staining; the supplied sources identify high-staining cells but no CAMLG-specific retrieval condition (HPA: cerebral cortex and fallopian tube, High).	Check section integrity and assay controls, then review retrieval, antibody dilution and detection against the antibody's IHC instructions (general IHC practice). Avoid assigning the failure to fixation without target-specific evidence.
The section shows widespread, uniform colour.	Background from detection activity, excess reagent exposure or inadequate washing can mask a cellular pattern (general IHC practice).	Compare with a no-primary section and inspect background outside expected-positive cells; adjust blocking, washes or detection exposure according to the assay controls (general IHC practice).
Reported not-detected cells stain strongly.	Signal in adipocytes, skeletal myocytes or soft-tissue fibroblasts conflicts with their reported HPA IHC categories; cross-reactivity or endogenous activity is possible (HPA: Not detected; general IHC practice).	Confirm the cell identity on the counterstained section, compare a no-primary control, and check whether expected-positive cells retain the reported cytoplasmic pattern (general IHC practice; HPA: tissue IHC profile).
Staining is chiefly nuclear.	Nuclear-dominant IHC conflicts with HPA's general cytoplasmic tissue profile, although uncertain nuclear locations appear in its separate ICC-IF summary (HPA: tissue IHC; HPA: subcellular ICC-IF).	Interpret the paraffin section against tissue IHC controls and review staining specificity before assigning a nuclear localisation; the uncertain ICC-IF finding does not validate nuclear-dominant IHC (HPA: subcellular ICC-IF; general IHC practice).
A low-staining tissue is used as the only positive control.	A weak control may be hard to distinguish from assay failure: parathyroid glandular cells and liver cholangiocytes are reported Low (HPA: tissue IHC profile).	Include a reported high-staining cell population, such as cerebral cortex neuronal cells or fallopian tube glandular cells, when available (HPA: High in both; general IHC practice).
Positive cells differ in intensity across tissues.	The reference profile itself spans High, Medium, Low and Not detected categories in specified cell populations (HPA: tissue IHC profile).	Score the named cell type and cytoplasmic localisation within each section before comparing intensity; investigate a mismatch against the relevant HPA category and assay controls (HPA: general cytoplasmic expression; general IHC practice).

Source: UniProt P49069 Human Protein Atlas tissue Human Protein Atlas subcellular

Experiment evidence

IHC & IF images showing CAMLG staining

IHC analysis of CAMLG using anti-CAMLG antibody (A08322-2). CAMLG was detected in a paraffin-embedded section of human breast 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-CAMLG Antibody (A08322-2) 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.

A08322-2 · IHCIHC analysis of CAMLG using anti-CAMLG antibody (A08322-2). CAMLG was detected in a paraffin-embedded section of human breast cancer tissue.

IHC analysis of CAMLG using anti-CAMLG antibody (A08322-2). CAMLG was detected in a paraffin-embedded section of mouse brain 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-CAMLG Antibody (A08322-2) 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.

A08322-2 · IHCIHC analysis of CAMLG using anti-CAMLG antibody (A08322-2). CAMLG was detected in a paraffin-embedded section of mouse brain tissue. Heat me

HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000164615-CAMLG/tissue/cerebral+cortex#imid_14962332

HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000164615-CAMLG/tissue/colon#imid_14962242

HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000164615-CAMLG/tissue/liver#imid_14962251

HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000164615-CAMLG/tissue/kidney#imid_14962252

HPA · ICC-IF HeLaHuman Protein Atlas immunofluorescence, HeLa, antibody HPA052636; green = CAMLG, blue = nucleus, red = microtubules.

← swipe / scroll for more images →

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

Sample controls for CAMLG IHC & IF

🧪Run cerebral cortex first and assess staining in neuronal cells (HPA: High in cerebral cortex neuronal cells). Use skeletal muscle myocytes as the negative tissue (HPA: Not detected in skeletal muscle myocytes); on the cortex slide, internal negative cells should show counterstain without specific chromogenic deposit, but their identity and negative status must be verified rather than assumed from the HPA row.

Positive control tissue: Cerebral cortex (Neuronal cells, HPA High)

Negative control tissue: Adipose tissue (HPA Not detected)

ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show CAMLG in HeLa, MCF-7, U2OS, ASC52telo, hTERT-RPE1 (serum starved), with annotated localisation: Vesicles (uncertain) (HPA subcellular).

Technical controls: Include a no-primary, secondary-only control and a host- and clonality-matched rabbit isotype control (selected-SKU IHC caption: rabbit primary antibody). Use CAMLG knockout material or a validated peptide-block control as a biological negative; quench endogenous peroxidase for chromogenic detection and assess neuronal pigment that could be mistaken for signal (standard IHC practice).

⚠️Feasibility: The target-specific fixation window and fixation effect are unreported in the supplied evidence; the selected-SKU paraffin-section caption does not state a fixative. Heat retrieval in EDTA at pH 8.0 is reported for the catalog antibody, but a retrieval dependency is unproven (selected-SKU IHC caption). The evidence does not establish that frozen sections or IF are easier; cerebral cortex pigment warrants careful interpretation of chromogenic signal (standard IHC practice).

HPA tissue IHC evidence for CAMLG

Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — 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
Cerebral cortex	Neuronal cells	High	Protein (IHC)	HPA →
Fallopian tube	Glandular cells	High	Protein (IHC)	HPA →
Adrenal gland	Glandular cells	Medium	Protein (IHC)	HPA →
Appendix	Glandular cells	Medium	Protein (IHC)	HPA →
Breast	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 →
Bone marrow	Hematopoietic cells	Not detected	Protein (IHC)	HPA →
Skeletal muscle	Myocytes	Not detected	Protein (IHC)	HPA →
Soft tissue	Fibroblasts	Not detected	Protein (IHC)	HPA →

Section 3

Advanced CAMLG IHC Tips

Use the IHC-validated antibody’s paraffin-section result as a starting point, then judge staining against CAMLG’s reported distribution and membrane topology.

Which retrieval condition should I start with for CAMLG paraffin sections?

Start with heat-mediated retrieval in EDTA at pH 8.0 (datasheet A08322-2). The selected paraffin-section image used this condition before overnight incubation at 4°C with 2 μg/ml catalog antibody (datasheet A08322-2). If staining is weak, first check heating consistency, section adhesion and reagent coverage, then compare a less or more intensive retrieval cycle on matched sections (standard IHC practice). Judge any adjustment by retained tissue morphology and reduced background as well as target signal (standard IHC practice). Keep retrieval identical across sections destined for quantitative comparison, and include the same positive-control tissue in each run (standard IHC practice).

How should I troubleshoot fixation-related changes in CAMLG staining?

The selected CAMLG image identifies paraffin-embedded tissue but does not state its fixative, so target-specific fixation sensitivity is unknown (datasheet A08322-2). Record each specimen’s fixative, fixation duration and processing history before comparing staining intensity (standard IHC practice). On matched sections, assess whether differences track processing batches, damaged morphology or uneven staining, while holding retrieval, antibody incubation and detection constant (standard IHC practice). The documented antibody incubation is 2 μg/ml overnight at 4°C; it provides a reproducible starting condition, not evidence for a particular fixative (datasheet A08322-2). Report unresolved fixation differences as a limitation of cross-specimen comparisons (standard IHC practice).

Where should convincing CAMLG chromogenic staining appear?

Expect predominantly cytoplasmic staining in tissue sections (HPA: general cytoplasmic expression), consistent with CAMLG’s endoplasmic-reticulum membrane assignment (UniProt P49069: subcellular location). Its 3 transmembrane segments occupy residues 190–207, 213–231 and 270–288, so a diffuse nuclear-only pattern needs independent validation (UniProt P49069: topology). Vesicular staining is plausible, although the reported vesicle, nucleoplasm and nucleoli IF locations are marked uncertain (HPA: subcellular). Compare staining with section morphology and a no-primary control before assigning puncta or nuclear signal to CAMLG (standard IHC practice). Score cytoplasmic and unexpected nuclear staining separately so an unusual compartment cannot inflate the primary readout (standard IHC practice).

How could CAMLG topology affect epitope access in paraffin sections?

CAMLG has no annotated isoforms in the supplied record, so an apparent staining shift should not be assigned to an isoform without additional evidence (UniProt P49069: isoforms). Its residues 1–189 and 232–269 face the cytoplasm, while 208–212 and 289–296 face the ER lumen (UniProt P49069: topology). If the antibody’s epitope is disclosed, map it to these intervals when interpreting retrieval-dependent staining (UniProt P49069: topology; standard IHC practice). No epitope location is supplied here, so keep that mapping conditional and compare matched retrieval conditions rather than assuming accessibility (datasheet A08322-2: epitope not stated; standard IHC practice). A reported phosphoserine at position 55 does not establish that this antibody distinguishes phosphorylation states (UniProt P49069: modified residues).

How can I cross-check CAMLG IHC with multiplex immunofluorescence?

Use IF as an orthogonal check of cell identity and compartment, while keeping chromogenic paraffin-section IHC as this guide’s primary readout (standard IHC practice). Pair CAMLG with a validated neuronal marker in cerebral cortex or a glandular-cell marker in fallopian tube, where those cell classes show high tissue IHC staining (HPA: tissue IHC). Choose fluorophores after inspecting unstained tissue autofluorescence, and include single-stain controls to identify spectral bleed-through (standard IF practice). Match permeabilisation to the mapped antibody epitope: CAMLG has cytoplasmic and short ER-lumen-facing regions, but this antibody’s epitope is unspecified (UniProt P49069: topology; datasheet A08322-2: epitope not stated). Treat nuclear IF signal cautiously because HPA labels those locations uncertain (HPA: subcellular).

What should I adjust if CAMLG DAB staining is widespread?

First compare a no-primary section with the test section to separate detection background from antibody-associated staining (standard IHC practice). The selected paraffin-section demonstration used 10% goat serum, 2 μg/ml primary antibody overnight at 4°C, an anti-rabbit peroxidase secondary for 30 minutes at 37°C, and DAB (datasheet A08322-2). Include a peroxidase-block step and inspect DAB development time as general chromogenic IHC controls (standard IHC practice). If both controls and specimens show diffuse pigment, troubleshoot blocking, washing and detection before attributing it to CAMLG (standard IHC practice). Assess tissue edges and damaged areas separately, since staining there can be misleading (standard IHC practice).

How should I score CAMLG across paraffin-section samples? ⚠ ANSWER MARKED FOR VERIFICATION

Define the cell population and compartment before scoring, because HPA reports general cytoplasmic expression and cell-type-dependent intensity (HPA: tissue IHC). For chromogenic sections, record the percentage of positive target cells and an intensity-based H-score using the same threshold and imaging conditions throughout (standard IHC practice). Normalise positive cells to all evaluable cells of the prespecified type, or report positive-cell density per mm² of viable tissue when cellularity varies (standard IHC practice). Exclude folds, necrosis and tissue edges by a predefined rule, and score unexpected nuclear signal separately (standard IHC practice). Include a common control section across batches to detect shifts in staining or DAB development (standard IHC practice).

How do I distinguish plausible CAMLG staining from artefact?

A plausible result is predominantly cytoplasmic in the relevant cells, given the tissue IHC profile and ER membrane assignment (HPA: general cytoplasmic expression; UniProt P49069: subcellular location). High staining in cerebral-cortex neuronal cells or fallopian-tube glandular cells offers reference patterns, while the selected product image shows staining in paraffin-embedded human breast cancer tissue (HPA: tissue IHC; datasheet A08322-2). Treat nuclear-only staining, strong signals confined to section edges or necrosis, and pigment reproduced in no-primary controls as reasons to investigate artefact (UniProt P49069: topology; standard IHC practice). Endogenous peroxidase can mimic DAB signal, so verify the peroxidase block and control section (standard IHC practice). HPA rates tissue staining Approved with medium RNA agreement; interpret discordant samples cautiously (HPA: reliability).

Boster reagents

Best CAMLG / Guided entry of tail-anchored proteins factor CAMLG IHC Antibodies

The IHC-validated anti-CAMLG antibody has real paraffin-section images from human breast cancer tissue and mouse and rat brain (catalog: IHC image captions); no IF image is supplied (catalog: IF images).

Real IHC data

Anti-CAMLG Antibody ®

Cat # A08322-2

View product →

A08322-2 is the sole card and lists IHC for human, mouse, and rat (catalog: applications and reactivity). Its images show paraffin-section staining in human breast cancer tissue and mouse and rat brain (catalog: IHC image captions).

Which to pick: Choose A08322-2 for paraffin-section IHC, including cross-species work, because its own captions document human, mouse, and rat staining (catalog: IHC image captions); the fixative is unreported (catalog: IHC image captions). No IF/ICC choice is supported because A08322-2 has no listed IF/ICC application or IF image (catalog: applications and IF images).

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

References

UniProt Consortium. UniProt entry P49069 (CAMLG_HUMAN, Guided entry of tail-anchored proteins factor CAMLG).
Human Protein Atlas. CAMLG tissue IHC expression (reliability: Approved).
Human Protein Atlas. CAMLG subcellular location (ICC-IF): Mainly localized to vesicles. In addition localized to the nucleoplasm and nucleoli..
Human Protein Atlas. CAMLG antibody validation summary (2 antibodies).
Calcium modulating ligand confers risk for Parkinson's disease and impacts lysosomes. Annals of clinical and translational neurology 2025 — PMC12093335.
PubMed PMID:7522304 — UniProt-cited evidence.
PubMed PMID:14702039 — UniProt-cited evidence.
PubMed PMID:15489334 — UniProt-cited evidence.