Anti-PIGA antibody (ab69768)

Overview

  • Product nameAnti-PIGA antibody
    See all PIGA primary antibodies
  • Description
    Rabbit polyclonal to PIGA
  • Tested applicationsSuitable for: ICC/IF, WBmore details
  • Species reactivity
    Reacts with: Human
    Predicted to work with: Mouse, Dog, Chimpanzee
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 200 - 300 of Human PIGA.

    (Peptide available as ab69767.)

  • Positive control
    • 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)

Properties

  • FormLiquid
  • 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 bufferPreservative: 0.02% Sodium Azide
    Constituents: 1% BSA, PBS, pH 7.4
  • Concentration information loading...
  • PurityImmunogen affinity purified
  • ClonalityPolyclonal
  • IsotypeIgG
  • Research areas

Applications

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.

Application Abreviews Notes
ICC/IF Use a concentration of 1 - 5 µg/ml.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 54 kDa (predicted molecular weight: 54 kDa).

Target

  • FunctionNecessary for the synthesis of N-acetylglucosaminyl-phosphatidylinositol, the very early intermediate in GPI-anchor biosynthesis.
  • PathwayGlycolipid biosynthesis; glycosylphosphatidylinositol-anchor biosynthesis.
  • Involvement in diseaseParoxysmal nocturnal hemoglobinuria (PNH) [MIM:300818]: A disorder characterized by hemolytic anemia with hemoglobinuria, thromboses in large vessels, and a deficiency in hematopoiesis. Clinical manifestation of red blood cell breakdown with release of hemoglobin into the urine is manifested most prominently by dark-colored urine in the morning. Note=The disease is caused by mutations affecting the gene represented in this entry.
    Multiple congenital anomalies-hypotonia-seizures syndrome 2 (MCAHS2) [MIM:300868]: An X-linked recessive developmental disorder characterized by dysmorphic features, neonatal hypotonia, myoclonic seizures, and variable congenital anomalies involving the central nervous, cardiac, and urinary systems. Most affected individuals die in infancy. Note=The disease is caused by mutations affecting the gene represented in this entry.
  • Sequence similaritiesBelongs to the glycosyltransferase group 1 family. Glycosyltransferase 4 subfamily.
  • Cellular localizationEndoplasmic reticulum membrane.
  • Information by UniProt
  • Database links
  • Alternative names
    • Class A GlcNAc inositol phospholipid assembly protein antibody
    • GlcNAc PI synthesis protein antibody
    • GlcNAc-PI synthesis protein antibody
    • GPI3 antibody
    • MCAHS2 antibody
    • Phosphatidylinositol glycan anchor biosynthesis class A antibody
    • Phosphatidylinositol glycan biosynthesis class A protein antibody
    • Phosphatidylinositol N-acetylglucosaminyltransferase subunit A antibody
    • Phosphatidylinositol-glycan biosynthesis class A protein antibody
    • PIG A antibody
    • PIG-A antibody
    • Piga antibody
    • PIGA_HUMAN antibody
    see all

Anti-PIGA antibody images

  • 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.

References for Anti-PIGA antibody (ab69768)

ab69768 has not yet been referenced specifically in any publications.

Product Wall

Thank you very much for your call earlier this week and for your patience while I've been in touch with the lab about the trouble with ab69768.

The lab used a protocol very similar to yours, andwe could find no record of any unusual steps req...

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (White blood cells)
Specification White blood cells
Fixative Formaldehyde
Permeabilization Yes - 0.1% Triton X-100
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Apr 09 2012

Application Immunocytochemistry/ Immunofluorescence
Sample Mouse Cell (bone maroow)
Specification bone maroow
Fixative Methanol
Permeabilization No
Blocking step Serum as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Jan 25 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (293 FT cells)
Loading amount 80 µg
Specification 293 FT cells
Gel Running Conditions Reduced Denaturing
Blocking step Milk as blocking agent for 9 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
Username

Abcam user community

Verified customer

Submitted Sep 17 2010

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"