Key features and details
- Mouse monoclonal [MEM-43/5] to CD59
- Suitable for: ICC/IF, IHC-P, WB, IP, Flow Cyt
- Reacts with: Mouse, Human
- Isotype: IgG2b
Product nameAnti-CD59 antibody [MEM-43/5]
See all CD59 primary antibodies
DescriptionMouse monoclonal [MEM-43/5] to CD59
SpecificityCD59 antigen (human). MEM-43/5 reacts with well defined epitope (around L33) and does not compete with MEM-43 and many other CD59 antibodies
Tested applicationsSuitable for: ICC/IF, IHC-P, WB, IP, Flow Cytmore details
Species reactivityReacts with: Mouse, Human
Thymocytes and T lymphocytes.
EpitopeThe antibody MEM-43/5 reacts with well defined epitope around L33 (see Bodian et al)
- Flow cyt: blood Jeg3 cell line IF/ICC
Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
Storage bufferpH: 7.40
Preservative: 0.097% Sodium azide
Concentration information loading...
PurityProtein A purified
Purification notesPurity >95% by SDS-PAGE.
Light chain typeunknown
Our Abpromise guarantee covers the use of ab9183 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|ICC/IF||Use a concentration of 5 µg/ml.|
|IHC-P||Use a concentration of 5 µg/ml.|
|WB||Use a concentration of 1 - 2 µg/ml. Use under non reducing condition. CD59 is GPI-anchored, so we recommend to use a laurylmatoside based lysis buffer or triton base buffer (see Bodian et al; 1% Triton X-100, 1 µg/ml leupeptin, 1 µg/ml pepstatin A and 1 mM phenlymethylsulphonyl fluoride in PBS), not NP40.|
|IP||Use at an assay dependent concentration.|
|Flow Cyt||Use a concentration of 1 µg/ml.
ab170192 - Mouse monoclonal IgG2b, is suitable for use as an isotype control with this antibody.
FunctionPotent inhibitor of the complement membrane attack complex (MAC) action. Acts by binding to the C8 and/or C9 complements of the assembling MAC, thereby preventing incorporation of the multiple copies of C9 required for complete formation of the osmolytic pore. This inhibitor appears to be species-specific. Involved in signal transduction for T-cell activation complexed to a protein tyrosine kinase.
The soluble form from urine retains its specific complement binding activity, but exhibits greatly reduced ability to inhibit MAC assembly on cell membranes.
Involvement in diseaseDefects in CD59 are the cause of CD59 deficiency (CD59D) [MIM:612300].
Sequence similaritiesContains 1 UPAR/Ly6 domain.
modificationsN- and O-glycosylated. The N-glycosylation mainly consists of a family of biantennary complex-type structures with and without lactosamine extensions and outer arm fucose residues. Also significant amounts of triantennary complexes (22%). Variable sialylation also present in the Asn-43 oligosaccharide. The predominant O-glycans are mono-sialylated forms of the disaccharide, Gal-beta-1,3GalNAc, and their sites of attachment are probably on Thr-76 and Thr-77. The GPI-anchor of soluble urinary CD59 has no inositol-associated phospholipid, but is composed of seven different GPI-anchor variants of one or more monosaccharide units. Major variants contain sialic acid, mannose and glucosamine Sialic acid linked to an N-acetylhexosamine-galactose arm is present in two variants.
Glycated. Glycation is found in diabetic subjects, but only at minimal levels in nondiabetic subjects. Glycated CD59 lacks MAC-inhibitory function and confers to vascular complications of diabetes.
Cellular localizationCell membrane. Secreted. Soluble form found in a number of tissues.
- Information by UniProt
- 16.3A5 antibody
- 1F5 antibody
- 1F5 antigen antibody
ICC/IF image of ab9183 stained Jeg3 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab9183, 5µg/ml) overnight at +4°C. The secondary antibody (green) was ab96879, DyLight® 488 goat anti-mouse IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
IHC image of ab9183 staining CD59 in Human normal placenta formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with EDTA (pH9, epitope retrieval solution 2) for 20 mins. The section was then incubated with ab9183, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
Overlay histogram showing Jurkat cells stained with ab9183 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab9183, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG2b [PLPV219] (ab91366, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in Jurkat cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.
ab9183 has been referenced in 9 publications.
- Zhou Y et al. CD59 is a potential biomarker of esophageal squamous cell carcinoma radioresistance by affecting DNA repair. Cell Death Dis 9:887 (2018). PubMed: 30166523
- Cupino TL et al. Stability and bioactivity of chitosan as a transfection agent in primary human cell cultures: A case for chitosan-only controls. Carbohydr Polym 180:376-384 (2018). PubMed: 29103517
- Kitterer D et al. Alteration of membrane complement regulators is associated with transporter status in patients on peritoneal dialysis. PLoS One 12:e0177487 (2017). PubMed: 28542228
- Reales E et al. The MAL protein is crucial for proper membrane condensation at the ciliary base, which is required for primary cilium elongation. J Cell Sci 128:2261-70 (2015). PubMed: 25967552
- Bromfield EG et al. The impact of oxidative stress on chaperone-mediated human sperm-egg interaction. Hum Reprod 30:2597-613 (2015). PubMed: 26345691
- Cai H et al. Differential transformation capacity of Src family kinases during the initiation of prostate cancer. Proc Natl Acad Sci U S A 108:6579-84 (2011). WB ; Human . PubMed: 21464326
- Renner B et al. The complement inhibitors Crry and factor H are critical for preventing autologous complement activation on renal tubular epithelial cells. J Immunol 185:3086-94 (2010). Flow Cyt . PubMed: 20675597
- Stechly L et al. Galectin-4-regulated delivery of glycoproteins to the brush border membrane of enterocyte-like cells. Traffic 10:438-50 (2009). WB . PubMed: 19192249
- Bodian DL et al. Mutational analysis of the active site and antibody epitopes of the complement-inhibitory glycoprotein, CD59. J Exp Med 185:507-16 (1997). PubMed: 9053451