Phospho-ATF2 (T71) Recombinant Rabbit Monoclonal Antibody [SC05-90]
cat.: ET1610-30
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat
Applications: WB, IF-Cell, IF-Tissue, IP, IHC-P
Clonality: Monoclonal
Clone number: SC05-90
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.
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: 55 kDa
Isotype: IgG
Immunogen: Synthetic phospho-peptide corresponding to residues surrounding Thr71 of Human ATF2 aa 58-94 / 505.
Positive control: Hela, NIH/3T3, SH-SY5Y, rat brain tissue, human prostate tissue, human uterus tissue, mouse colon tissue, human cervix tissue, human skin tissue, human breast tissue, human esophagus tissue, human placenta tissue, mouse large intestine tissue, Hela cells treated with 250ng/mL anisomycin for 30 minutes.
Subcellular location: Mitochondrion outer membrane, Nucleus, Cytoplasm.
Recommended Dilutions:
  WB
  IF-Cell
  IF-Tissue
  IHC-P
1:1,000
1:50-1:200
1:50-1:200
1:100-1:800
Uniprot #: SwissProt: P15336 Human | P16951 Mouse | Q00969 Rat
Alternative names: Activating transcription factor 2 Activating transcription factor 2 splice variant ATF2 var2 ATF 2 Atf-2 Atf2 ATF2 protein ATF2_HUMAN cAMP Response Element Binding Protein 2 cAMP response element binding protein CRE BP1 cAMP response element-binding protein CRE-BP1 cAMP responsive element binding protein 2, formerly cAMP-dependent transcription factor ATF-2 cAMP-responsive element-binding protein 2 CRE BP1 CRE-BP CREB 2 CREB-2 CREB2 CREBP1 Cyclic AMP dependent transcription factor ATF 2 Cyclic AMP-dependent transcription factor ATF-2 Cyclic AMP-responsive element-binding protein 2 D130078H02Rik D18875 HB 16 HB16 Histone acetyltransferase ATF2 MGC105211 MGC105222 MGC111558 MGC142504 mXBP MXBP protein Tg(Gzma-Klra1)7Wum TREB 7 TREB7
Images
ET1610-30_1.jpg Fig1: ICC staining of Phospho-ATF2 (T71) in Hela cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1610-30, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
ET1610-30_2.jpg Fig2: ICC staining of Phospho-ATF2 (T71) in NIH/3T3 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1610-30, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
ET1610-30_3.jpg Fig3: ICC staining of Phospho-ATF2 (T71) in SH-SY5Y cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1610-30, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
ET1610-30_4.jpg Fig4: Immunohistochemical analysis of paraffin-embedded rat brain tissue with Rabbit anti-Phospho-ATF2 (T71) antibody (ET1610-30) at 1/800 dilution.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1610-30) at 1/800 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ET1610-30_5.jpg Fig5: Immunohistochemical analysis of paraffin-embedded human prostate tissue using anti-Phospho-ATF2 (T71) antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 20 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1610-30, 1/200) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ET1610-30_6.jpg Fig6: Immunohistochemical analysis of paraffin-embedded human uterus tissue using anti-Phospho-ATF2 (T71) antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 20 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1610-30, 1/200) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ET1610-30_7.jpg Fig7: Immunohistochemical analysis of paraffin-embedded mouse colon tissue using anti-Phospho-ATF2 (T71) antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 20 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1610-30, 1/200) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ET1610-30_8.jpg Fig8: Immunohistochemical analysis of paraffin-embedded human cervix tissue with Rabbit anti-Phospho-ATF2 (T71) antibody (ET1610-30) at 1/200 dilution.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1610-30) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ET1610-30_9.jpg Fig9: Immunohistochemical analysis of paraffin-embedded human skin tissue with Rabbit anti-Phospho-ATF2 (T71) antibody (ET1610-30) at 1/200 dilution.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1610-30) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ET1610-30_10.jpg Fig10: Immunohistochemical analysis of paraffin-embedded human breast tissue with Rabbit anti-Phospho-ATF2 (T71) antibody (ET1610-30) at 1/200 dilution.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1610-30) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ET1610-30_11.jpg Fig11: Immunohistochemical analysis of paraffin-embedded human esophagus tissue with Rabbit anti-Phospho-ATF2 (T71) antibody (ET1610-30) at 1/200 dilution.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1610-30) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ET1610-30_12.jpg Fig12: Immunohistochemical analysis of paraffin-embedded human placenta tissue with Rabbit anti-Phospho-ATF2 (T71) antibody (ET1610-30) at 1/200 dilution.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1610-30) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ET1610-30_13.jpg Fig13: Immunohistochemical analysis of paraffin-embedded mouse large intestine tissue with Rabbit anti-Phospho-ATF2 (T71) antibody (ET1610-30) at 1/800 dilution.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1610-30) at 1/800 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ET1610-30_14.jpg Fig14: Immunocytochemistry analysis of Hela cells treated with or without 250ng/mL anisomycin for 30 minutes labeling Phospho-ATF2 (T71) with Rabbit anti-Phospho-ATF2 (T71) antibody (ET1610-30) at 1/50 dilution.

Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-Phospho-ATF2 (T71) antibody (ET1610-30) at 1/50 dilution in 2% negative goat serum 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.
ET1610-30_15.jpg Fig15: Western blot analysis of Phospho-ATF2 (T71) on HeLa cell lysates treated with or without untreated anisomycin(250ng/ml 30min) with Rabbit anti-Phospho-ATF2 (T71) antibody (ET1610-30) at 1/1,000 dilution.

Lysates/proteins at 20 µg/Lane.

Predicted band size: 70 kDa
Observed band size: 70 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 (ET1610-30) at 1/1,000 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.
ET1610-30_16.jpg Fig16: Western blot analysis of Phospho-ATF2 (T71) on NIH/3T3 cell lysates treated with or without untreated anisomycin(250ng/ml 30min) with Rabbit anti-Phospho-ATF2 (T71) antibody (ET1610-30) at 1/1,000 dilution.

Lysates/proteins at 20 µg/Lane.

Predicted band size: 70 kDa
Observed band size: 70 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 (ET1610-30) at 1/1,000 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.
ET1610-30_17.jpg Fig17: Phospho-ATF2 (T71) was immunoprecipitated in 0.2mg HeLa cell treated anisomycin(25ug/ml 30min) lysate with (ET1610-30) at 2 µg/25 µl agarose. Western blot was performed from the immunoprecipitate using (ET1610-30) at 1/1,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody (NBI01H) at 1/5,000 dilution was used for 1 hour at room temperature.

Lane 1: HeLa cell lysate (input)
Lane 2: (ET1610-30) IP in HeLa cell lysate
Lane 3: Rabbit IgG instead of (ET1610-30) in HeLa cell lysate

Blocking/Dilution buffer: 5% NFDM/TBST
Exposure time: 3 mintues
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.