GADD45A Rabbit Polyclonal Antibody

• Catalog Number: orb10683
• Category: Antibodies
• Description: GADD45A Rabbit Polyclonal Antibody
• Species/Host: Rabbit
• Clonality: Polyclonal
• Tested applications: FC, ICC, IF, IHC-Fr, IHC-P, WB
• Predicted Reactivity: Bovine, Gallus, Porcine
• Reactivity: Human, Mouse, Rat
• Isotype: IgG
• Immunogen: KLH conjugated synthetic peptide derived from human GADD45 (65-165/165aa)
• Antibody Type: Primary Antibody
• Concentration: 1mg/ml
• Dilution range: WB=1:500-2000, IHC-P=1:100-500, IHC-F=1:100-500, ICC/IF=1:100-500, IF=1:100-500, Flow-Cyt=3ug/Test
• Form/Appearance: Liquid
• Conjugation: Unconjugated
• MW: 18 kDa
• Target: GADD45A
• UniProt ID: P24522
• RRID: AB_10762846
• Storage: Maintain refrigerated at 2-8°C for up to 2 weeks. For long term storage store at -20°C in small aliquots to prevent freeze-thaw cycles.
• Buffer/Preservatives: 0.01M TBS (pH7.4) with 1% rAlbumin, 0.02% Proclin300 and 50% Glycerol.
• Alternative names: Growth Arrest And DNA Damage Inducible alpha; DNA
• Note: For research use only
• Expiration Date: 12 months from date of receipt.

Blank control: A431. Primary Antibody (green line): Rabbit Anti-GADD45 antibody (orb10683), Dilution: 1 µg/10^6 cells, Isotype Control Antibody (orange line): Rabbit IgG. Secondary Antibody: Goat anti-rabbit IgG-AF647, Dilution: 1 µg/Test. Protocol, The cells were fixed with 4% PFA (10 min at room temperature) and then permeabilized with 0.1% PBST for 20 min at room temperature. The cells were then incubated in 5% BSA to block non-specific protein-protein interactions for 30 min at room temperature. Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20000 events was performed.

Paraformaldehyde-fixed, paraffin embedded (Human liver carcinoma), Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min, Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes, Blocking buffer (normal goat serum) at 37°C for 30 min, Antibody incubation with (GADD45) Polyclonal Antibody, Unconjugated (orb10683) at 1:400 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.

Paraformaldehyde-fixed, paraffin embedded (Mouse brain), Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min, Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes, Blocking buffer (normal goat serum) at 37°C for 30 min, Antibody incubation with (GADD45) Polyclonal Antibody, Unconjugated (orb10683) at 1:400 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.

Sample: Lane 1: Brain (Rat) Lysate at 30 ug, Lane 2: Kidney (Rat) Lysate at 30 ug, Primary: Anti-GADD45 (orb10683) at 1:200 dilution, Secondary: HRP conjugated Goat-Anti-Rabbit IgG (orb572747) at 1:3000 dilution, Predicted band size: 18kD, Observed band size: 18kD, We are unsure as to the identity of these extra bands.

Tissue/Cell: A431 cell, 4% Paraformaldehyde-fixed, Triton X-100 at room temperature for 20 min, Blocking buffer (normal goat serum) at 37°C for 20 min, Antibody incubation with (GADD45) Polyclonal Antibody, Unconjugated (orb10683) 1:200, 90 minutes at 37°C, followed by a conjugated Goat Anti-Rabbit IgG antibody (orb868805) at 37°C for 90 minutes, DAPI (5 ug/ml, blue) was used to stain the cell nuclei.

Tissue/Cell: rat brain tissue, 4% Paraformaldehyde-fixed and paraffin-embedded, Antigen retrieval: citrate buffer (0.01M, pH6.0), Boiling bathing for 15 min, Block endogenous peroxidase by 3% Hydrogen peroxide for 30 min, Blocking buffer (normal goat serum) at 37°C for 20 min, Incubation: Anti-GADD45 Polyclonal Antibody, Unconjugated (orb10683) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody and DAB staining.

Tissue/Cell: U-2OS cell, 4% Paraformaldehyde-fixed, Triton X-100 at room temperature for 20 min, Blocking buffer (normal goat serum) at 37°C for 20 min, Antibody incubation with (GADD45) Polyclonal Antibody, Unconjugated (orb10683) 1:200, 90 minutes at 37°C, followed by a conjugated Goat Anti-Rabbit IgG antibody (orb868805) at 37°C for 90 minutes, DAPI (5 ug/ml, blue) was used to stain the cell nuclei.