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ab69767 |
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Read our guarantee »Products:Cardiovascular >> Blood >> Erythrocytes
Anti-PIGA antibody
Rabbit polyclonal to PIGA
ICC/IF, WBmore details
Reacts with
Human
Predicted to work with
Mouse, Dog, Chimpanzee
Synthetic peptide conjugated to KLH derived from within residues 200 - 300 of Human PIGA.
(Peptide available as ab69767.)
This antibody gave a positive signal in the following Human Whole Cell Lysates: TE 671, A498, HeLa - Hydroxyurea Treated (48hr, 2µM), Jurkat - Staurosporine Treated (24hr, 500nM), Y79, HeLa - Staurosporine Treated (24hr, 500nM), HeLa - Hydroxyurea Treated (48hr, 1µM)
Liquid
Store at +4°C short term (1-2 weeks). Aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
Preservative: 0.02% Sodium Azide
Constituents: 1% BSA, PBS, pH 7.4
Concentration information loading...
Immunogen affinity purified
Polyclonal
IgG
Cardiovascular >> Blood >> Erythrocytes
Our Abpromise guarantee covers the use of ab69768 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 1 - 5 µg/ml.
WB: Use a concentration of 1 µg/mlDetects a band of approximately 54 kDa (predicted molecular weight: 54 kDa).
PIGA belongs to the glycosyltransferase 1 family and is necessary for the synthesis of N-acetylglucosaminyl-phosphatidylinositol, the very early intermediate in GPI-anchor biosynthesis. Defects in PIGA are the cause of paroxysmal nocturnal hemoglobinuria (PNH) which is an acquired hemolytic blood disorder characterized by chronic hemolysis with hemoglobinuria, increased tendency to venous thrombosis, and variable degrees of bone marrow failure.
Endoplasmic reticulum membrane; Single-pass membrane protein.
Western blot - PIGA antibody (ab69768)

All lanes : Anti-PIGA antibody (ab69768) at 1 µg/ml
Lane 1 : TE 671 (Human Rhabdomyosarcoma) Whole Cell Lysate
Lane 2 : A498 (Human Kidney Carcinoma) Whole Cell Lysate
Lane 3 : HeLa Whole Cell Lysate - Hydroxyurea Treated (48hr, 2��M)
Lane 4 : Jurkat Whole Cell Lysate - Staurosporine Treated (24hr, 500nM)
Lane 5 : Y79 (Human retinoblastoma cell line) Whole Cell Lysate
Lane 6 : HeLa Whole Cell Lysate - Staurosporine Treated (24hr, 500nM)
Lane 7 : HeLa Whole Cell Lysate - Hydroxyurea Treated (48hr, 1µM)
Lysates/proteins at 10 µg per lane.
Secondary
Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
Performed under reducing conditions.
Predicted band size : 54 kDa
Observed band size : 54 kDa
Additional bands at : 75 kDa. We are unsure as to the identity of these extra bands.
Immunocytochemistry/ Immunofluorescence - PIGA antibody (ab69768)

ICC/IF image of ab69768 stained HeLa cells. The cells were 4% PFA 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 (ab69768, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 Goat anti-Rabbit IgG (H+L) used at a 1/1000 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. This antibody also gave a positive result in 4% PFA fixed (10 min) HepG2 cells at 1µg/ml, and in 100% Methanol fixed (5 min) HeLa, Hek293, HepG2, MCF-7 cells at 5µg/ml.
ab69768 has not yet been referenced specifically in any publications.
Publishing research using ab69768? Please let us know so that we can cite the reference in this datasheet
Concentration of lot no. is
Concentration not available for this lot.
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All lanes : Anti-PIGA antibody (ab69768) at 1 µg/ml
Lane 1 : TE 671 (Human Rhabdomyosarcoma) Whole Cell Lysate
Lane 2 : A498 (Human Kidney Carcinoma) Whole Cell Lysate
Lane 3 : HeLa Whole Cell Lysate - Hydroxyurea Treated (48hr, 2µM)
Lane 4 : Jurkat Whole Cell Lysate - Staurosporine Treated (24hr, 500nM)
Lane 5 : Y79 (Human retinoblastoma cell line) Whole Cell Lysate
Lane 6 : HeLa Whole Cell Lysate - Staurosporine Treated (24hr, 500nM)
Lane 7 : HeLa Whole Cell Lysate - Hydroxyurea Treated (48hr, 1µM)
Lysates/proteins at 10 µg per lane.
Secondary
Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
Performed under reducing conditions.
Predicted band size : 54 kDa
Observed band size : 54 kDa
Additional bands at : 75 kDa. We are unsure as to the identity of these extra bands.

ICC/IF image of ab69768 stained HeLa cells. The cells were 4% PFA 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 (ab69768, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 Goat anti-Rabbit IgG (H+L) used at a 1/1000 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. This antibody also gave a positive result in 4% PFA fixed (10 min) HepG2 cells at 1µg/ml, and in 100% Methanol fixed (5 min) HeLa, Hek293, HepG2, MCF-7 cells at 5µg/ml.
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