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CAMLG / Guided entry of tail-anchored proteins factor CAMLG · IHC design guide
Design Immunohistochemistry for CAMLG
Plan CAMLG paraffin IHC using its cytoplasmic tissue staining pattern (HPA tissue IHC) and ER membrane location (UniProt). The catalog antibody has an IHC dilution range of 2–5 μg/ml (datasheet A08322-2); cerebral cortex neurons stain strongly, while adipocytes have no detected staining (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
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 stain (HPA tissue IHC); ER membrane protein (UniProt) | |
| Staining pattern | Cytoplasmic in 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 conditions consistent across sections. (standard IHC practice; not target-specific) | |
| Caveat | Staining has medium consistency with tissue RNA data (HPA tissue IHC) | |
| Regulation | Highest in brain, testis and ovary (UniProt) | |
| Isoform / epitope | No annotated isoforms; cytoplasmic vs lumenal epitope matters (UniProt) |
Section 1
Recommended CAMLG IHC & IF Protocols
The catalog antibody’s IHC-P protocol uses EDTA pH 8.0 retrieval (datasheet A08322-2). The published CAMLG brain-section protocol below uses heated citric acid retrieval (PMC12093335: methods).
| 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 EDTA pH 8.0 for the catalog antibody (datasheet A08322-2); consider citric acid retrieval when adapting the published brain-section protocol (PMC12093335: methods).
Section 2
What Is the Expected CAMLG Staining Pattern?
CAMLG is an endoplasmic reticulum membrane protein with 3 transmembrane segments (UniProt P49069 topology). In paraffin-section IHC, expect predominantly cytoplasmic staining, especially in cerebral cortex neuronal cells and fallopian tube glandular cells (HPA: general cytoplasmic expression; High in both cell populations). Interpret the pattern with caution: HPA rates its tissue IHC evidence Approved, with medium consistency between staining and RNA expression (HPA: tissue IHC reliability).
| Cytoplasmic chromogen in cortex neurons or fallopian tube glandular cells, stronger than nearby background. | This matches the reported High staining in those cells and the general cytoplasmic IHC profile (HPA: tissue IHC). An ER membrane protein can appear as cytoplasmic staining at light-microscope resolution (UniProt P49069: ER membrane). Assess intensity and distribution in the named cells, rather than treating every stained cell as equally informative. |
| Predominantly nuclear or sharply plasma-membrane staining, with little cytoplasmic signal. | That distribution conflicts with the reported tissue IHC profile and ER membrane annotation (HPA: general cytoplasmic expression; UniProt P49069: ER membrane). Consider nonspecific staining or an interpretation artefact; inspect morphology and controls before calling it CAMLG. HPA's uncertain nuclear ICC-IF observations do not establish a nuclear IHC pattern (HPA: subcellular ICC-IF). |
| Strong signal in adipocytes, marrow hematopoietic cells, skeletal myocytes, or soft-tissue fibroblasts. | HPA reports CAMLG as Not detected in those particular cell populations (HPA: tissue IHC). Investigate antibody cross-reactivity or endogenous detection activity, especially if the positive control stains as expected. These are comparison cells, not universal biological negatives: UniProt describes CAMLG expression as ubiquitous (UniProt P49069: tissue specificity). |
| Diffuse chromogen across cells and extracellular spaces, obscuring cell boundaries. | The distribution cannot be scored confidently against HPA's cell-specific, general cytoplasmic profile (HPA: tissue IHC). In routine IHC, incomplete blocking, excess antibody, or detection background can produce diffuse signal (general IHC practice). Compare a no-primary control and review the detection steps before interpreting weak cellular staining. |
| No detectable staining in cortex neurons or fallopian tube glandular cells. | This misses the strongest supplied tissue examples (HPA: High in both cell populations). First check section integrity and a working positive control; then review retrieval, antibody concentration, and detection as general IHC variables. A failed positive control prevents a reliable negative call for the test section (general IHC practice). |
💡Expected CAMLG appearanceA convincing positive is predominantly cytoplasmic chromogen in cortex neurons or fallopian tube glandular cells, where HPA reports High staining; isolated nuclear, surface-only, or diffuse extracellular signal is suspect (HPA: tissue IHC; UniProt P49069: ER membrane).
| Cell population and tissue | HPA reports High staining in cortex neuronal cells and fallopian tube glandular cells, Medium staining in several other listed populations, and Not detected in specified adipocytes, marrow cells, myocytes, and fibroblasts (HPA: tissue IHC). Score the named cell population rather than assigning one intensity to the whole section. |
| Membrane topology and epitope access | CAMLG has 3 transmembrane segments and both cytoplasmic and lumenal regions (UniProt P49069 topology). An antibody's epitope location is not supplied, so topology alone cannot justify a particular retrieval method or predict how processing will affect staining. |
| IHC evidence and antibody choice | HPA lists HPA052636 and HPA056472 as IHC Approved, and rates the tissue profile Approved with medium staining–RNA consistency (HPA: antibodies; tissue IHC reliability). Approved supports use as a starting point, while the observed pattern and controls still determine whether an individual run is interpretable. |
| Low and absent HPA observations | HPA reports Low staining in selected glandular cells, cholangiocytes, and splenic red-pulp cells, while some other listed populations are Not detected (HPA: tissue IHC). Weak or absent staining in one population should be compared with its own HPA entry; UniProt's ubiquitous tissue description does not imply detectable IHC in every cell (UniProt P49069: tissue specificity). |
| ICC-IF localisation evidence | HPA calls vesicles the main ICC-IF location and nucleoplasm and nucleoli additional locations, all uncertain (HPA: subcellular ICC-IF). These observations may guide a separate IF assessment, but they do not override the general cytoplasmic tissue IHC profile or establish a nuclear paraffin-IHC positive call (HPA: tissue IHC). |
| Situation | Likely cause | Next action |
|---|---|---|
| Known-positive cortex neurons or fallopian tube glandular cells are blank. | The run may have insufficient detectable signal; HPA reports High staining in these cells (HPA: tissue IHC). | Confirm the positive-control section and detection reagents worked, then review retrieval and antibody concentration as general IHC checks. Do not interpret test-section absence while the control fails (general IHC practice). |
| A broad brown haze makes cytoplasmic staining hard to score. | Background from blocking, antibody concentration, or detection can obscure the reported cytoplasmic pattern (general IHC practice; HPA: tissue IHC profile). | Inspect a no-primary control and review blocking, washes, antibody concentration, and development time (general IHC practice). Score CAMLG only where cellular signal can be distinguished from background. |
| Nuclear staining dominates the paraffin section. | Predominant nuclear signal conflicts with the ER membrane annotation and general cytoplasmic tissue profile (UniProt P49069; HPA: tissue IHC). | Review morphology and controls, and compare the signal with cytoplasmic staining in a positive tissue. HPA's nuclear ICC-IF locations are uncertain and do not validate a nuclear IHC call (HPA: subcellular ICC-IF). |
| A purported negative comparator stains strongly. | HPA's Not detected calls apply to named cell populations, while UniProt describes broad tissue expression (HPA: tissue IHC; UniProt P49069: tissue specificity). | Verify which cells actually stain. If the signal is in an HPA Not detected population, check a no-primary control for endogenous detection activity and consider antibody cross-reactivity (general IHC practice). |
| Results differ between IHC antibodies or sections. | Both listed antibodies are IHC Approved, but HPA reports only medium consistency between antibody staining and RNA expression for the tissue profile (HPA: antibodies; tissue IHC reliability). | Compare the same named cell populations and matched controls across sections. Treat disagreement as unresolved until the staining distribution and controls support an interpretation (general IHC practice). |
| Q: Should an IF/ICC image show the same pattern as paraffin IHC? | HPA reports vesicular ICC-IF staining with additional nuclear locations, each uncertain, alongside a general cytoplasmic tissue IHC profile (HPA: subcellular ICC-IF; tissue IHC). | A: Evaluate IF/ICC localisation on its own guide page and retain the uncertainty labels. Do not use those ICC-IF observations to redefine a positive chromogenic IHC result (HPA: subcellular ICC-IF; tissue IHC). |
Experiment evidence
IHC & IF images showing CAMLG staining
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.
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.
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 require staining in neuronal cells (HPA: High in cerebral cortex neuronal cells). Use adipose tissue as a negative comparator (HPA: Not detected in adipocytes); on the positive slide, assess adjacent non-neuronal cells as internal negatives only where they show no primary-dependent staining, since their CAMLG status is not specified by the supplied HPA rows.
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 concentration-matched rabbit IgG isotype control matched to the primary antibody’s clonality once confirmed (selected-SKU caption: rabbit primary; clonality unreported). Use CAMLG knockout material as a biological specificity control where available; for cerebral cortex IHC, quench endogenous peroxidase and check the no-primary slide for pigment that could resemble DAB staining (standard IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the fixative used for the selected-SKU paraffin section is unreported (selected-SKU caption: paraffin section; fixative not stated). The documented IHC procedure uses heat retrieval in EDTA at pH 8.0, so verify retrieval performance for the chosen preparation (selected-SKU caption: heat retrieval in EDTA, pH 8.0). The supplied evidence does not establish whether frozen sections or IF are easier; in cerebral cortex, distinguish tissue pigment from chromogenic signal with the no-primary control (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
Troubleshoot CAMLG staining in paraffin sections using the catalog antibody’s IHC evidence, expected cellular pattern, and matched controls.
Which antigen retrieval should I try first for CAMLG IHC?
Start with heat-mediated retrieval in EDTA at pH 8.0 for this catalog antibody (datasheet A08322-2). The documented paraffin-section example used this retrieval before 2 μg/ml primary antibody overnight at 4°C; its fixative and retrieval heating duration were not reported (caption A08322-2). If staining is weak, first check that heating, cooling, and section coverage are consistent, then compare a small retrieval-time series while keeping detection conditions fixed (standard IHC practice). Score cellular cytoplasmic staining against a same-run positive control, because diffuse DAB throughout the section does not establish CAMLG-specific retrieval (HPA: general cytoplasmic expression; standard IHC practice).
How can I troubleshoot fixation without knowing CAMLG’s fixation sensitivity?
CAMLG-specific sensitivity to fixation is unknown from the supplied evidence, and the catalog paraffin-section caption does not state a fixative (caption A08322-2). Record each specimen’s actual fixative, fixation duration, and processing history, then compare sections processed alike before attributing a staining difference to CAMLG biology (standard IHC practice). Use the documented EDTA retrieval at pH 8.0 as a consistent starting condition and keep the 2 μg/ml primary concentration constant during that comparison (datasheet A08322-2). Include a same-run positive section and a no-primary control so weak staining and fixation-related background can be assessed separately (standard IHC practice).
What staining compartment should count as a plausible CAMLG signal?
Expect predominantly cytoplasmic staining in tissue sections, consistent with the reported general IHC pattern and CAMLG’s endoplasmic reticulum membrane annotation (HPA: general cytoplasmic expression; UniProt P49069: endoplasmic reticulum membrane). Its 3 transmembrane segments lie at residues 190–207, 213–231, and 270–288, so a perinuclear or fine cytoplasmic pattern is plausible, although ordinary DAB IHC cannot establish an ER membrane location by itself (UniProt P49069 topology; standard IHC practice). HPA cell imaging also reports vesicles as the main location, with nuclear locations marked uncertain (HPA subcellular: uncertain locations). Treat isolated nuclear-only DAB staining cautiously and compare it with cytoplasmic signal, controls, and tissue morphology (HPA tissue IHC; standard IHC practice).
How should unknown epitope placement affect CAMLG IHC troubleshooting?
The supplied record annotates 0 isoforms and does not specify the catalog antibody’s epitope, so a staining difference cannot be assigned to an isoform or mapped to a particular residue (UniProt P49069 record; caption A08322-2). CAMLG’s cytoplasmic regions span residues 1–189 and 232–269, while short lumenal regions span 208–212 and 289–296 (UniProt P49069 topology). When signal is weak, vary retrieval before claiming that one membrane-facing epitope is inaccessible, and keep section thickness and detection conditions comparable (standard IHC practice). The annotated phosphoserine at 55 establishes a modification site, but without an epitope map or phospho-specific validation it does not explain an IHC result (UniProt P49069 modified residues; caption A08322-2).
How can IF help check a puzzling chromogenic CAMLG pattern?
Use IF as a separate corroborating experiment: HPA cell imaging places CAMLG mainly in vesicles, while its additional nucleoplasmic and nucleolar calls are uncertain (HPA subcellular). For tissue multiplexing, pair CAMLG with a marker validated for the cell population being scored, such as the neuronal population reported as high in cerebral cortex, and inspect each channel independently (HPA tissue IHC: cerebral cortex neuronal cells high; standard IF practice). Choose fluorophores and filter sets after checking unstained tissue autofluorescence, and include single-color controls before interpreting overlap (standard IF practice). Because this antibody’s epitope is unspecified, optimize permeabilisation cautiously for access to the relevant cytoplasmic or lumenal side; the documented IHC conditions do not establish IF performance (UniProt P49069 topology; caption A08322-2).
What should I change when CAMLG DAB staining is diffuse?
First compare a no-primary section with the stained section to separate detection-system color from antibody-associated staining (standard IHC practice). The documented paraffin example used 10% goat serum, 2 μg/ml primary antibody overnight at 4°C, and a peroxidase-conjugated secondary for 30 minutes at 37°C before DAB development (caption A08322-2). If background persists, verify the general peroxidase block, shorten DAB development, and check wash consistency one variable at a time (standard IHC practice). Judge improvement by retention of cellular cytoplasmic signal and reduced color in no-primary and tissue-background areas, since the reported tissue pattern is generally cytoplasmic (HPA tissue IHC; standard IHC practice).
How should I score CAMLG staining across paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and cytoplasmic compartment before scoring, because HPA reports general cytoplasmic staining with different levels across cell types (HPA tissue IHC). For chromogenic sections, record the percentage of positive target cells and intensity bins, then calculate an H-score if those bins are applied consistently (standard IHC practice). Alternatively, report positive cells per mm² of viable tissue and normalize comparisons to the number of eligible cells or measured viable area, rather than total section area alone (standard IHC practice). Hold acquisition, DAB development, and scoring thresholds constant across batches, and document excluded necrotic or folded regions (standard IHC practice).
When is an apparent CAMLG-positive cell likely to be artefactual?
A convincing result shows cellular cytoplasmic staining in morphologically intact cells and exceeds the matched no-primary background (HPA: general cytoplasmic expression; standard IHC practice). Strong nuclear-only color is less persuasive because HPA’s nucleoplasmic and nucleolar imaging calls are uncertain, while UniProt places CAMLG at the endoplasmic reticulum membrane (HPA subcellular; UniProt P49069: endoplasmic reticulum membrane). Check whether staining concentrates at section edges, folds, or necrotic regions, and whether it remains after accounting for endogenous peroxidase in the DAB workflow (standard IHC practice). Interpret a negative cell population in context: HPA reports high neuronal staining in cerebral cortex but no detected staining in adipocytes, and its overall tissue IHC reliability is approved with medium RNA agreement (HPA tissue IHC).
Boster reagents
Best CAMLG / Guided entry of tail-anchored proteins factor CAMLG IHC Antibodies
A08322-2 has IHC images from paraffin sections of human breast cancer, mouse brain, and rat brain (catalog image captions). No IF/ICC data are supplied (catalog applications and images).
Real IHC data
Anti-CAMLG Antibody ®
Cat # A08322-2
A08322-2 is the only listed SKU; its application list includes IHC and its stated reactivity covers Human, Mouse, and Rat (catalog applications/reactivity). Its IHC images show human breast cancer, mouse brain, and rat brain paraffin sections (catalog image captions).
Which to pick: Choose A08322-2 for paraffin-section tissue IHC; its own captions document staining at 2 μg/ml in the pictured tissues (catalog image captions). For cross-species work, A08322-2 is a rabbit antibody with Human, Mouse, and Rat reactivity (catalog host/reactivity). No listed SKU has IF/ICC validation (catalog applications and images), and the IHC captions do not report the fixative (catalog image captions).
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.
Immunohistochemistry protocol sheet for CAMLG (Guided entry of tail-anchored proteins factor CAMLG). Expected localisation Cytoplasmic tissue stain (HPA tissue IHC); ER membrane protein (UniProt). Antigen retrieval: EDTA pH 8.0 HIER, heat-mediated (datasheet A08322-2). Validated IHC antibody: A08322-2 at 2-5 μg/ml (the product's own IHC validation image is shown). Controls (HPA): positive Cerebral cortex (Neuronal cells); negative Adipose tissue. IHC-P workflow on paraffin sections; fixative unspecified. Protocol steps: 1. Sample: Paraffin-embedded human breast cancer tissue; fixative not specified (datasheet A08322-2); 2. Fixation: Image fixative and duration unreported (datasheet A08322-2); verify before use.; 3. Sectioning: 4–5 µm sections on charged slides (standard); 4. Deparaffinisation: Xylene, graded ethanol series to water (standard); 5. Antigen retrieval: Heat retrieval: EDTA pH 8.0 (datasheet A08322-2); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% goat serum (datasheet A08322-2); 8. Primary antibody: Rabbit anti-CAMLG, 2-5 μg/ml (datasheet A08322-2); 9. Primary incubation: Overnight at 4 °C (datasheet A08322-2). Watch for: Fixation: Keep fixation conditions consistent across sections. (standard IHC practice; not target-specific); Caveat: Staining has medium consistency with tissue RNA data (HPA tissue IHC); Isoform / epitope: No annotated isoforms; cytoplasmic vs lumenal epitope matters (UniProt); Regulation: Highest in brain, testis and ovary (UniProt). Evidence: UniProt P49069, HPA tissue IHC · Approved, HPA ICC-IF · Mainly localized to vesicles. In addition localized to the…, Datasheet A08322-2, PMC12093335. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/CAMLG/immunohistochemistry-ihc-guide.