GGA3 antibody

• Catalog Number: orb345520
• Category: Antibodies
• Description: GGA3 antibody
• Species/Host: Rabbit
• Clonality: Polyclonal
• Tested applications: ELISA, IP, WB
• Reactivity: Human, Primate
• Isotype: IgG
• Immunogen: This affinity-purified antibody was prepared from whole rabbit serum produced by repeated immunizations with a synthetic peptide corresponding to an internal region near amino acids 400-425 of Human GGA3.
• Concentration: 1.0mg/mL
• Dilution range: ELISA: 1:10,000 - 1:50,000, IP: 1:100, WB: 1:500 - 1:3,000
• Form/Appearance: Liquid (sterile filtered)
• Purity: This affinity purified antibody is directed against human GGA3 protein. The product was affinity purified from monospecific antiserum by immunoaffinity purification. A BLAST analysis was used to suggest reactivity with this protein from human and chimpanzee based on 100% homology for the immunogen sequence. Cross reactivity may occur with GGA3 from macaque (93% homology) and orangutan (87% homology) sources. The immunizing sequence is present on both the long (isoform 1) and short (isoform 2) transcript variants for this protein.
• Conjugation: Unconjugated
• UniProt ID: Q9NZ52
• NCBI: 20336267
• Storage: Store vial at -20° C prior to opening. Aliquot contents and freeze at -20° C or below for extended storage. Avoid cycles of freezing and thawing. Centrifuge product if not completely clear after standing at room temperature. This product is stable for several weeks at 4° C as an undiluted liquid. Dilute only prior to immediate use.
• Buffer/Preservatives: 0.01% (w/v) Sodium Azide
• Alternative names: rabbit anti-GGA3 antibody, GGA-3, GGA 3, ADP-ribos
• Note: For research use only
• Application notes: This affinity purified antibody has been tested for use in ELISA and western blot. Specific conditions for reactivity should be optimized by the end user. Expect bands at 78 kDa in size corresponding to GGA3 by western blotting in the appropriate cell lysate or extract.
• Expiration Date: 12 months from date of receipt.

ELISA results of purified Rabbit anti-GGA3 Antibody tested against BSA-conjugated peptide of immunizing peptide. Each well was coated in duplicate with 0.1 µg of conjugate. The starting dilution of antibody was 5 µg/ml and the X-axis represents the Log10 of a 3-fold dilution. This titration is a 4-parameter curve fit where the IC50 is defined as the titer of the antibody. Assay performed using 3% fish gel, Goat anti-Rabbit IgG Antibody Peroxidase Conjugated (Min X Bv Ch Gt GP Ham Hs Hu Ms Rt & Sh Serum Proteins) (p/n orb347654) and TMB ELISA Peroxidase Substrate (p/n orb348651).

Western Blot of Rabbit anti-GGA3 antibody. Marker: Opal Pre-stained ladder. Lane 1: HEK293 lysate (p/n orb348669). Lane 2: HeLa Lysate (p/n orb348668). Lane 3: MCF-7 Lysate (p/n orb348664). Lane 4: Jurkat Lysate. Lane 5: A431 Lysate (p/n orb348665). Lane 6: LNCaP Lysate (p/n orb348694). Lane 7: A-172 Lysate (p/n orb348708). Lane 8: NIH/3T3 Lysate (p/n orb348714). Load: 35 µg per lane. Primary antibody: GGA3 antibody at 1:5000 for overnight at 4°C. Secondary antibody: Peroxidase rabbit secondary antibody (p/n orb347654) at 1:30000 for 60 min at RT. Blocking Buffer: 1% Casein-TTBS for 30 min at RT. Predicted/Observed size: 78 kDa for GGA3.

Western blot using Biorbyt's Affinity Purified anti-GGA3 antibody shows detection of a band at ~110 kDa corresponding to GFP-GGA3 fusion protein present in a lysate of HEK293 cells over- expressing the recombinant protein (lane 2, arrowhead). Pre-incubation of antibody with immunizing peptide blocks specific staining (lane 3). MW markers are shown in lane 1 (700 nm channel - red). Approximately 35 µg of lysate was separated on a 16% Tricine gel by SDS-PAGE and transferred onto nitrocellulose. After blocking the membrane was probed with the primary antibody diluted to 1:600. Reaction occurred overnight at 4°C followed by washes and reaction with a 1:10000 dilution of IRDye800 conjugated Gt-a-Rabbit IgG [H&L] for 45 min at room temperature (800 nm channel - green).