ATF4 Recombinant Rabbit Monoclonal Antibody [SD20-92]
cat.: ET1612-37
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat
Applications: WB, IF-Cell, IF-Tissue, IHC-P, IP, FC
Clonality: Monoclonal
Clone number: SD20-92
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: 39 kDa
Isotype: IgG
Immunogen: Recombinant protein within Human ATF4 aa 1-220 / 351.
Positive control: Hela cell lysate, PC-12 cell lysate, HL-60 cell lysate, K562 cell lysate, human lung carcinoma tissue lysate, N2A, human liver carcinoma tissue, human prostate carcinoma tissue, human skin tissue, human breast carcinoma tissue, human stomach carcinoma tissue, human small intestine tissue, human colon carcinoma tissue, Hela, mouse testis tissue lysate.
Subcellular location: Nucleus, Nucleus speckle, Cell membrane, Cytoplasm, Centrosome.
Recommended Dilutions:
  WB
  IF-Cell
  IF-Tissue
  IHC-P
  FC
  IP
1:500-1:2,000
1:50-1:200
1:50-1:200
1:50-1:1,000
1:50-1:100
Use at an assay dependent concentration.
Uniprot #: SwissProt: P18848 Human | Q06507 Mouse | Q9ES19 Rat
Alternative names: Activating transcription factor 4 ATF 4 ATF4 ATF4 protein ATF4_HUMAN cAMP-dependent transcription factor ATF-4 cAMP-responsive element-binding protein 2 CREB 2 CREB-2 CREB2 Cyclic AMP dependent transcription factor ATF 4 Cyclic AMP response element binding protein 2 Cyclic AMP-dependent transcription factor ATF-4 Cyclic AMP-responsive element-binding protein 2 DNA binding protein TAXREB67 DNA-binding protein TAXREB67 Tax Responsive Enhancer Element B67 Tax-responsive enhancer element-binding protein 67 TaxREB67 TXREB
Images
ET1612-37_1.jpg Fig1: Western blot analysis of ATF4 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 (ET1612-37, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.
Positive control:
Lane 1: Hela cell lysate
Lane 2: PC-12 cell lysate
Lane 3: HL-60 cell lysate
Lane 4: K562 cell lysate
Lane 5: human lung carcinoma tissue lysate
ET1612-37_2.jpg Fig2: ICC staining of ATF4 in N2A 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 (ET1612-37, 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).
ET1612-37_3.jpg Fig3: Immunohistochemical analysis of paraffin-embedded human liver carcinoma tissue using anti-ATF4 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 (ET1612-37, 1/50) 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.
ET1612-37_4.jpg Fig4: Immunohistochemical analysis of paraffin-embedded human thyroid tissue using anti-ATF4 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 (ET1612-37, 1/50) 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.
ET1612-37_5.jpg Fig5: Immunohistochemical analysis of paraffin-embedded human prostate carcinoma tissue using anti-ATF4 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 (ET1612-37, 1/50) 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.
ET1612-37_6.jpg Fig6: Immunohistochemical analysis of paraffin-embedded human skin tissue using anti-ATF4 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 (ET1612-37, 1/50) 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.
ET1612-37_7.jpg Fig7: Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue using anti-ATF4 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 (ET1612-37, 1/50) 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.
ET1612-37_8.jpg Fig8: Immunohistochemical analysis of paraffin-embedded human stomach carcinoma tissue using anti-ATF4 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 (ET1612-37, 1/50) 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.
ET1612-37_9.jpg Fig9: Immunohistochemical analysis of paraffin-embedded human small intestine tissue using anti-ATF4 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 (ET1612-37, 1/50) 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.
ET1612-37_10.jpg Fig10: Immunohistochemical analysis of paraffin-embedded mouse colon tissue using anti-ATF4 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 (ET1612-37, 1/50) 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.
ET1612-37_11.jpg Fig11: Immunohistochemical analysis of paraffin-embedded human colon carcinoma tissue with Rabbit anti-ATF4 antibody (ET1612-37) at 1/1,000 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 (ET1612-37) at 1/1,000 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.
ET1612-37_12.jpg Fig12: Flow cytometric analysis of ATF4 was done on Hela cells. The cells were fixed, permeabilized and stained with the primary antibody (ET1612-37, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Rabbit IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
ET1612-37_13.jpg Fig13: Western blot analysis of ATF4 on mouse testis tissue lysates with Rabbit anti-ATF4 antibody (ET1612-37) at 1/500 dilution.

Lysates/proteins at 20 µg/Lane.

Predicted band size: 39 kDa
Observed band size: 55 kDa

Exposure time: 2 minutes;

10% 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 (ET1612-37) at 1/500 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:300,000 dilution was used for 1 hour at room temperature.
ET1612-37_14.jpg Fig14: Immunohistochemical analysis of paraffin-embedded rat
colon tissue with Rabbit anti-ATF4 antibody (ET1612-37) 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 (ET1612-37) 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.
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