SMPD2 Antibody, FITC conjugated
Description
Introduction to SMPD2 and FITC-Conjugated Antibodies
SMPD2 (sphingomyelin phosphodiesterase 2) is a neutral sphingomyelinase that hydrolyzes sphingomyelin into ceramide and phosphocholine. It plays critical roles in cellular signaling, apoptosis, and membrane remodeling . The FITC-conjugated SMPD2 antibody is a fluorescently labeled immunoglobulin designed to detect and visualize SMPD2 protein in cells. FITC (fluorescein isothiocyanate) enables real-time localization studies via microscopy, making it essential for immunofluorescence (IF) and immunocytochemistry (ICC) .
Cell Cycle Regulation
SMPD2 knockdown (KD) induces G1 phase arrest by upregulating cyclin-dependent kinase inhibitors (P21, P27) and reducing phosphorylated Akt levels, impairing PI3K/Akt signaling . FITC-conjugated antibodies are pivotal in tracking SMPD2 localization during cell cycle progression.
ER Stress and Cellular Fitness
SMPD2 KD impairs the unfolded protein response (UPR) under ER stress, reducing LAMP1 mRNA/protein levels without affecting lysosomal acidification. This highlights SMPD2’s role in buffering ER stress .
Signaling Pathway Modulation
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Wnt Signaling: SMPD2 KD reduces β-catenin and Wnt3a levels, disrupting canonical Wnt signaling .
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PI3K/Akt Pathway: Reduced phosphorylated Akt levels upon SMPD2 KD correlate with diminished cell proliferation .
Immunofluorescence (IF) Protocol
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Primary Antibody: Incubate overnight with FITC-conjugated SMPD2 antibody (e.g., 5–20 µg/ml for Abbexa) .
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Secondary Detection: FITC-conjugated secondary antibodies (e.g., 1:200 dilution) .
Western Blot (WB) Protocol
Limitations and Considerations
Product Specs
Constituents: 50% Glycerol, 0.01M PBS, pH 7.4
Target Background
- Interestingly, neutral sphingomyelinase (but not acid sphingomyelinase) was essential for PorBIA-mediated Neisseria gonorrhoeae invasion into host cells. PMID: 25224994
- nSMase1 (sphingomyelin phosphodiesterase 2, SMPD2) plays a central role in ceramide signaling during stress and cytokine responses, as well as apoptosis. PMID: 25168245
- These findings suggest that oxidized low-density lipoprotein (Ox-LDL) activates the N-SMase-ceramide signaling pathway, stimulating phosphorylation of p38 MAPK and inducing apoptosis in vascular smooth muscle cells, ultimately initiating vascular smooth muscle cell calcification. PMID: 24358176
- NSM2 plays a key role in the cellular response to hyperosmolar stress, regulating both cytokine secretion and lipid droplet formation. PMID: 22899568
- nSMase2 is involved in ceramide generation, aberrant apoptosis, and lung injury under cigarette smoke exposure. PMID: 20448054
- N-SMase at the cytofacial plasma membrane is critical for the proper orientation of polymorphonuclear leukocytes (PMNs) in formyl-methionyl-leucyl-phenylalanine (FMLP) gradients, at least partially through polarizing the distribution of Rac 1/2 and RhoA GTPases. PMID: 20378749
- Results indicate that WithaD enhances ceramide accumulation by activating N-SMase 2, modulates phosphorylation of the JNK and p38MAPK, and induces apoptosis in both myeloid and lymphoid cells, as well as primary cells derived from leukemia patients. PMID: 20836852
- Data demonstrate that dopamine significantly increased NSMase2 mRNA and protein levels, while minimal changes were observed in NSMase1 and NSMase3 mRNAs. PMID: 19698806
- nSMase2 functions as a growth suppressor in MCF7 cells, linking confluence to the G(0)/G(1) cell cycle checkpoint. PMID: 15051724
- Identification of 15 polymorphisms, including 9 novel single nucleotide polymorphisms (SNPs), in the SMPD2 gene; one haplotype showed associations with the extrinsic type of atopic dermatitis subgroup. PMID: 17212982
- The catalytic region of neutral sphingomyelinase 2 where palmitoylation occurs is found to be localized at the inner leaflet of the plasma membrane. PMID: 17272284
- This study highlights the importance of the neutral sphingomyelinase-ceramide pathway in mediating oxidative stress-induced apoptosis and cell death of human primary oligodendrocytes. PMID: 18040843
