HA722854

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:	Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
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

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.

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.

	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.
	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.
	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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