CREB Recombinant Rabbit Monoclonal Antibody [SA04-04]
cat.: ET1601-15
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Zebrafish, Rat
Applications: WB, IF-Cell, IF-Tissue, IHC-P, IP, FC
Clonality: Monoclonal
Clone number: SA04-04
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: 35 kDa
Isotype: IgG
Immunogen: Synthetic peptide within human CREB aa 250-290.
Positive control: HeLa cell lysate, A431 cell lysate, U-2 OS cell lysate, HEK-293 cell lysate, COS-1 cell lysate, NIH/3T3 cell lysate, C2C12 cell lysate, C6 cell lysate, PC-12 cell lysate, rat testis tissue lysate, mouse brain tissue lysate, rat brain tissue lysate, zebrafish tissue lysates, Hela, human tonsil tissue, human lung carcinoma tissue, human breast carcinoma tissue, human thyroid tissue, human stomach carcinoma tissue, human pancreas tissue, mouse colon tissue, rat brain tissue, NIH/3T3, C6, human cerebellum tissue, mouse cerebellum tissue, rat cerebellum tissue.
Subcellular location: Nucleus
Recommended Dilutions:
  WB
  IF-Cell
  IF-Tissue
  IHC-P
  FC
  IP
1:500-1:1,000
1:500
1:200
1:500-1:5,000
1:1,000
Use at an assay dependent concentration.
Uniprot #: SwissProt: P16220 Human | Q01147 Mouse | P15337 Rat
Alternative names: Active transcription factor CREB cAMP response element binding protein 1 cAMP response element binding protein cAMP responsive element binding protein 1 cAMP-responsive element-binding protein 1 CREB CREB-1 CREB1 CREB1_HUMAN Cyclic AMP-responsive element-binding protein 1 MGC9284 OTTHUMP00000163864 OTTHUMP00000163865 OTTHUMP00000206660 OTTHUMP00000206662 OTTHUMP00000206667 Transactivator protein
Images
ET1601-15_1.jpg Fig1: Western blot analysis of CREB on different lysates with Rabbit anti-CREB antibody (ET1601-15) at 1/2,000 dilution.

Lane 1: HeLa cell lysate (20 µg/Lane)
Lane 2: A431 cell lysate (20 µg/Lane)
Lane 3: U-2 OS cell lysate (20 µg/Lane)
Lane 4: HEK-293 cell lysate (20 µg/Lane)
Lane 5: COS-1 cell lysate (20 µg/Lane)
Lane 6: NIH/3T3 cell lysate (20 µg/Lane)
Lane 7: C2C12 cell lysate (20 µg/Lane)
Lane 8: C6 cell lysate (20 µg/Lane)
Lane 9: PC-12 cell lysate (20 µg/Lane)
Lane 10: Rat testis tissue lysate (40 µg/Lane)
Lane 11: Mouse brain tissue lysate (40 µg/Lane)
Lane 12: Rat brain tissue lysate (40 µg/Lane)

Predicted band size: 35 kDa
Observed band size: 42 kDa

Exposure time: 6 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 (ET1601-15) at 1/2,000 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.
ET1601-15_2.jpg Fig2: Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue using anti-CREB antibody.

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 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1601-15, 1/500) 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.
ET1601-15_3.jpg Fig3: Immunohistochemical analysis of paraffin-embedded human thyroid tissue using anti-CREB antibody.

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 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1601-15, 1/500) 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.
ET1601-15_4.jpg Fig4: Immunohistochemical analysis of paraffin-embedded human stomach carcinoma tissue using anti-CREB antibody.

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 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1601-15, 1/500) 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.
ET1601-15_5.jpg Fig5: Immunohistochemical analysis of paraffin-embedded human pancreas tissue using anti-CREB antibody.

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 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1601-15, 1/500) 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.
ET1601-15_6.jpg Fig6: Immunohistochemical analysis of paraffin-embedded mouse colon tissue using anti-CREB antibody.

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 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1601-15, 1/500) 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.
ET1601-15_7.jpg Fig7: Immunohistochemical analysis of paraffin-embedded rat brain tissue with Rabbit anti-CREB antibody (ET1601-15) 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 (ET1601-15) 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.
ET1601-15_8.jpg Fig8: Immunocytochemistry analysis of NIH/3T3 cells labeling CREB with Rabbit anti-CREB antibody (ET1601-15) at 1/500 dilution.

Cells were fixed in 100% precooled methanol 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-CREB antibody (ET1601-15) at 1/500 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.
ET1601-15_9.jpg Fig9: Immunocytochemistry analysis of C6 cells labeling CREB with Rabbit anti-CREB antibody (ET1601-15) at 1/500 dilution.

Cells were fixed in 100% precooled methanol 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-CREB antibody (ET1601-15) at 1/500 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.
ET1601-15_10.jpg Fig10: Immunohistochemical analysis of paraffin-embedded human cerebellum tissue with Rabbit anti-CREB antibody (ET1601-15) at 1/5,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 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 (ET1601-15) at 1/5,000 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.
ET1601-15_11.jpg Fig11: Immunohistochemical analysis of paraffin-embedded mouse cerebellum tissue with Rabbit anti-CREB antibody (ET1601-15) at 1/5,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 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 (ET1601-15) at 1/5,000 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.
ET1601-15_12.jpg Fig12: Immunohistochemical analysis of paraffin-embedded rat cerebellum tissue with Rabbit anti-CREB antibody (ET1601-15) at 1/5,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 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 (ET1601-15) at 1/5,000 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.
ET1601-15_13.jpg Fig13: Flow cytometric analysis of NIH/3T3 cells labeling CREB.

Cells were fixed and permeabilized. Then stained with the primary antibody (ET1601-15, 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).
ET1601-15_14.jpg Fig14: Flow cytometric analysis of C6 cells labeling CREB.

Cells were fixed and permeabilized. Then stained with the primary antibody (ET1601-15, 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).
ET1601-15_15.jpg Fig15: Western blot analysis of CREB on zebrafish tissue lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1601-15, 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.
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