What fixation and permeabilization protocol do you recommend for optimal lysosomal staining in IF?

To preserve the integrity of the lysosomal membrane, we recommend a gentle fixation and permeabilization procedure. Fix your cells with 4% paraformaldehyde (PFA) for 15 minutes at room temperature. For permeabilization, use a mild detergent like 50 µg/mL digitonin or 0.1% saponin for 5-10 minutes. This is generally better than using harsh detergents like Triton X-100, which can disrupt organelle membranes and cause the soluble lysosomal contents to leak out.

What is a good positive control cell line or tissue for ACP2?

ACP2 is a lysosomal enzyme, so cells with high lysosomal activity are excellent positive controls. We recommend using macrophage cell lines. For tissues, the liver, kidney, and spleen are rich in lysosomes and will provide a strong signal in both Western Blotting and IHC.

Mouse Anti-ACP2 Recombinant Antibody (V2-610934) (CBMAB-A0077-LY)

See all ACP2 antibodies

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Datasheet

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Basic Information

Host Animal

Mouse

Clone

V2-610934

Application

WB, ELISA

Immunogen

ACP2 (AAH03160, 1 a.a. ~ 423 a.a) full-length recombinant protein with GST tag. MW of the GST tag alone is 26 KDa.

Specificity

Human

Antibody Isotype

IgG2b, λ

Clonality

Monoclonal

Application Notes

The COA includes recommended starting dilutions, optimal dilutions should be determined by the end user.

Application	Note
IF(ICC)	10 µg/ml
IHC	5 µg/ml

Formulations & Storage [For reference only, actual COA shall prevail!]

Format

Liquid

Buffer

PBS, pH7.4

Preservative

None

Concentration

Batch dependent

Purity

> 95% Purity determined by SDS-PAGE.

Storage

Store at +4°C short term (1-2 weeks). Aliquot and store at -20°C long term. Avoid repeated freezethaw cycles.

More Infomation

Target

Full Name

acid phosphatase 2, lysosomal

Introduction

This gene encodes the beta subunit of lysosomal acid phosphatase (LAP). LAP is chemically and genetically distinct from red cell acid phosphatase. The encoded protein belongs to a family of distinct isoenzymes which hydrolyze orthophosphoric monoesters to alcohol and phosphate. Mutations in this gene or in the related alpha subunit gene cause acid phosphatase deficiency. Multiple alternatively spliced transcript variants encoding different isoforms have been identified for this gene. [provided by RefSeq]

Entrez Gene ID

Biological Process

Dephosphorylation
Lysosome organization

Cellular Location

Lysosome membrane; Lysosome lumen. The soluble form arises by proteolytic processing of the membrane-bound form.

Topology

Lumenal: 31-380 aa
Helical: 381-401 aa
Cytoplasmic: 402-423 aa

PTM

The membrane-bound form is converted to the soluble form by sequential proteolytic processing. First, the C-terminal cytoplasmic tail is removed. Cleavage by a lysosomal protease releases the soluble form in the lysosome lumen.
N-glycosylated. The intermediates formed during enzymatic deglycosylation suggest that all eight predicted N-glycosylation sites are used.

Jiao, X., Rahimi Balaei, M., Abu-El-Rub, E., Casoni, F., Pezeshgi Modarres, H., Dhingra, S., ... & Marzban, H. (2021). Reduced Granule Cell Proliferation and Molecular Dysregulation in the Cerebellum of Lysosomal Acid Phosphatase 2 (ACP2) Mutant Mice. International journal of molecular sciences, 22(6), 2994.

Mehdizadeh, M., Ashtari, N., Jiao, X., Rahimi Balaei, M., Marzban, A., Qiyami-Hour, F., ... & Marzban, H. (2020). Alteration of the Dopamine Receptors’ Expression in the Cerebellum of the Lysosomal Acid Phosphatase 2 Mutant (Naked–Ataxia (NAX)) Mouse. International journal of molecular sciences, 21(8), 2914.

de Pizzol Júnior, J. P., Sasso-Cerri, E., & Cerri, P. S. (2018). Matrix metalloproteinase-1 and acid phosphatase in the degradation of the lamina propria of eruptive pathway of rat molars. Cells, 7(11), 206.

Lee, J., Kim, J., Son, K., d’Alexandry, A. L. P. H., & Min, J. Y. (2017). Acid phosphatase 2 (ACP2) is required for membrane fusion during influenza virus entry. Scientific reports, 7(1), 1-13.

Lee, Y. C., Su, C. Y., Lin, Y. F., Lin, C. M., Fang, C. Y., Lin, Y. K., ... & Chen, C. L. (2017). Lysosomal acid phosphatase 2 is an unfavorable prognostic factor but is associated with better survival in stage II colorectal cancer patients receiving chemotherapy. Oncotarget, 8(7), 12120.

Ashtari, N. (2017). Cerebellar corticogenesis in the lysosomal acid phosphatase (acp2) mutant mice: purkinje cell migration disorder.

Jiao, X. (2017). The study of sonic hedgehog signaling pathway in the development of lysosomal acid phosphatase (Acp2) mutant mice cerebellar granule cells.

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Please try the standard protocols which include: protocols, troubleshooting and guide.

Enzyme-linked Immunosorbent Assay (ELISA)
Flow Cytometry
Immunofluorescence (IF)
Immunohistochemistry (IHC)
Immunoprecipitation (IP)
Western Blot (WB)
Enzyme-Linked Immunospot (ELISpot)
Proteogenomics
Other Protocols

Isotype control

For research use only. Not intended for any clinical use.

Custom Antibody Labeling

We also offer labeled antibodies developed using our catalog antibody products and nonfluorescent conjugates (HRP, AP, Biotin, etc.) or fluorescent conjugates (Alexa Fluor, FITC, TRITC, Rhodamine, Texas Red, R-PE, APC, Qdot Probes, Pacific Dyes, etc.).

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