DDIT3 Recombinant Rabbit Monoclonal Antibody [JM10-31]
cat.: ET1703-05
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat
Applications: WB, FC
Clonality: Monoclonal
Clone number: JM10-31
Form: Liquid
Storage condition: Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term.
Storage buffer: 1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration: 1ug/ul
Purification: Protein A affinity purified.
Molecular weight: Predicted band size: 19 kDa
Isotype: IgG
Immunogen: Synthetic peptide within Human DDIT3 aa 135-169 / 169.
Positive control: SW480 cell lysate, HeLa cell lysate, LOVO cell lysate, PC-12 cell lysate, Hela.
Subcellular location: Cytoplasm, Nucleus.
Recommended Dilutions:
  WB
  FC

1:5,000
1:50-1:100
Uniprot #: SwissProt: P35638 Human | P35639 Mouse | Q62857 Rat
Alternative names: C/EBP homologous protein C/EBP Homology Protein C/EBP zeta C/EBP-homologous protein 10 C/EBP-homologous protein CCAAT/enhancer binding protein homologous protein CEBPZ CHOP 10 CHOP CHOP-10 CHOP10 DDIT 3 DDIT-3 Ddit3 DDIT3_HUMAN DNA Damage Inducible Transcript 3 DNA damage-inducible transcript 3 protein GADD 153 GADD153 Growth Arrest and DNA Damage Inducible Protein 153 Growth arrest and DNA damage inducible protein GADD153 Growth arrest and DNA damage-inducible protein GADD153 MGC4154
Images
ET1703-05_1.jpg Fig1: Western blot analysis of DDIT3 on different lysates with Rabbit anti-DDIT3 antibody (ET1703-05) at 1/5,000 dilution.

Lane 1: SW480 cell lysate (20 µg/Lane)
Lane 2: HeLa cell lysate (20 µg/Lane)

Predicted band size: 19 kDa
Observed band size: 27 kDa

Exposure time: 3 minutes;

4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1703-05) at 1/5,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
ET1703-05_2.jpg Fig2: Western blot analysis of DDIT3 on different lysates with Rabbit anti-DDIT3 antibody (ET1703-05) at 1/1,000 dilution.

Lane 1: SW480-si NT cell lysate
Lane 2: SW480-si DDIT3 cell lysate

Lysates/proteins at 3 µg/Lane.

Predicted band size: 19 kDa
Observed band size: 25 kDa

Exposure time: 5 minutes;

4-20% SDS-PAGE gel.

ET1703-05 was shown to specifically react with DDIT3 in SW480-si NT cells. Weakened band was observed when SW480-si DDIT3 sample was tested. SW480-si NT and SW480-si DDIT3 samples were subjected to SDS-PAGE. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM in TBST for 1 hour at room temperature. The primary antibody (ET1703-05, 1/1,000) and Loading control antibody (Rabbit anti-GAPDH, ET1601-4, 1/10,000) were used in 5% BSA at room temperature for 2 hours. Goat Anti-rabbit IgG-HRP Secondary Antibody (HA1001) at 1:100,000 dilution was used for 1 hour at room temperature.
ET1703-05_3.jpg Fig3: Western blot analysis of DDIT3 on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1703-05, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:200,000 dilution was used for 1 hour at room temperature.
Positive control:
Lane 1: LOVO cell lysate
Lane 2: PC-12 cell lysate

Predicted band size: 19 kDa
Observed band size: 25 kDa
ET1703-05_4.jpg Fig4: Flow cytometric analysis of DDIT3 was done on Hela cells. The cells were fixed, permeabilized and stained with the primary antibody (ET1703-05, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor®488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.