DDIT3 Recombinant Rabbit Monoclonal Antibody [PSH07-65]
cat.: HA722854
| Product Type: | Recombinant Rabbit monoclonal IgG, primary antibodies |
|---|---|
| Species reactivity: | Human, Mouse, Rat |
| Applications: | WB, IF-Cell, FC |
| Clonality: | Monoclonal |
| Clone number: | PSH07-65 |
| 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: | PBS (pH7.4), 0.1% 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: | Recombinant protein within human DDIT3 aa 1-169. |
| Positive control: | HeLa treated with 2μg/mL tunicamycin for 8 hours cell lysate, C2C12 treated with 2μg/mL thapsigargin for 8 hours cell lysate, C6 cell lysate, C6 treated with 2μg/mL tunicamycin for 8 hours cell lysate, HeLa cells treated with 2μg/mL tunicamycin for 8 hours, C2C12 cells treated with 2μg/mL thapsigargin for 8 hours, C6 cells treated with 2μg/mL tunicamycin for 8 hours. |
| Subcellular location: | Cytoplasm, Nucleus. |
| Recommended Dilutions:
WB IF-Cell FC |
1:5,000 1:100 1:1,000 |
| 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
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Fig1:
Western blot analysis of DDIT3 on different lysates with Rabbit anti-DDIT3 antibody (HA722854) at 1/5,000 dilution. Lane 1: HeLa cell lysate Lane 2: HeLa treated with 2μg/mL tunicamycin for 8 hours cell lysate Lane 3: C2C12 cell lysate Lane 4: C2C12 treated with 2μg/mL thapsigargin for 8 hours cell lysate Lane 5: C6 cell lysate Lane 6: C6 treated with 2μg/mL tunicamycin for 8 hours cell lysate Lysates/proteins at 20 µg/Lane. Predicted band size: 19 kDa Observed band size: 27 kDa Exposure time: Lane 1-2: 3 minutes; Lane 3-6: 10 seconds; ECL: K1801; 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 (HA722854) 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. |
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Fig2:
Immunocytochemistry analysis of HeLa cells treated with 2μg/mL tunicamycin for 8 hours labeling DDIT3 with Rabbit anti-DDIT3 antibody (HA722854) at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-DDIT3 antibody (HA722854) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution. |
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Fig3:
Immunocytochemistry analysis of C2C12 cells treated with 2μg/mL thapsigargin for 8 hours labeling DDIT3 with Rabbit anti-DDIT3 antibody (HA722854) at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-DDIT3 antibody (HA722854) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution. |
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Fig4:
Immunocytochemistry analysis of C6 cells treated with 2μg/mL tunicamycin for 8 hours labeling DDIT3 with Rabbit anti-DDIT3 antibody (HA722854) at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-DDIT3 antibody (HA722854) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution. |
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Fig5:
Flow cytometric analysis of C2C12 cells treated with 2μg/mL thapsigargin for 8 hours labeling DDIT3. Cells were fixed and permeabilized. Then stained with the primary antibody (HA722854, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black). |
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