Notch 1 Recombinant Rabbit Monoclonal Antibody [SJ205]
cat.: ET1606-55
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: SJ205
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: 273 kDa
Isotype: IgG
Immunogen: Synthetic peptide within Human Notch 1 aa 2,481-2,530 / 2,555.
Positive control: MDA-MB-231 cell lysate, THP-1 cell lysate, human pancreas tissue, mouse lung tissue, mouse pancreas tissue, Hela cell.
Subcellular location: Cell membrane, Nucleus(intracellular domain).
Recommended Dilutions:
  WB
  IF-Cell
  IF-Tissue
  IHC-P
  FC
  IP
1:1,000-1:2,000
1:50-1:200
1:50-1:200
1:50-1:200
1:50-1:100
Use at an assay dependent concentration.
Uniprot #: SwissProt: P46531 Human | Q01705 Mouse
Alternative names: 9930111A19Rik AOS5 AOVD1 hN1 Lin-12 LIN12 Mis6 Motch A mT14 Neurogenic locus notch homolog protein 1 Neurogenic locus notch protein homolog NICD NOTC1_HUMAN Notch 1 Notch 1 intracellular domain NOTCH Notch gene homolog 1 (Drosophila) Notch homolog 1, translocation-associated (Drosophila) NOTCH, Drosophila, homolog of, 1 notch1 TAN1 Translocation associated notch homolog Translocation associated notch protein TAN 1 Translocation-associated notch protein TAN-1 xotch
Images
ET1606-55_1.jpg Fig1: Western blot analysis of Notch 1 on different lysates with Rabbit anti-Notch 1 antibody (ET1606-55) at 1/2,000 dilution.

Lane 1: MDA-MB-231 cell lysate
Lane 2: THP-1 cell lysate

Lysates/proteins at 15 µg/Lane.

Predicted band size: 273 kDa
Observed band size: 125 kDa

Exposure time: 3 minutes 10 seconds;

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 (ET1606-55) 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.
ET1606-55_2.jpg Fig2: Western blot analysis of Notch 1 on different lysates with Rabbit anti-Notch 1 antibody (ET1606-55) at 1/1,000 dilution.

Lane 1: Hela-si NT cell lysate
Lane 2: Hela-si Notch 1 cell lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 273 kDa
Observed band size: 125 kDa

Exposure time: 6 minutes;

4-20% SDS-PAGE gel.

ET1606-55 was shown to specifically react with Notch 1 in Hela-si NT cells. Weakened band was observed when Hela-si Notch 1 sample was tested. Hela-si NT and Hela-si Notch 1 samples were subjected to SDS-PAGE. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM in TBST for 1 hour at room temperature. The primary antibody (ET1606-55, 1/1,000) and Loading control antibody (Rabbit anti-GAPDH, ET1601-4, 1/10,000) were used in 5% BSA at room temperature for 2 hours. Goat Anti-rabbit IgG-HRP Secondary Antibody (HA1001) at 1:100,000 dilution was used for 1 hour at room temperature.
ET1606-55_3.jpg Fig3: ICC staining of Notch 1 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 (ET1606-55, 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).
ET1606-55_4.jpg Fig4: ICC staining of Notch 1 in HepG2 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 (ET1606-55, 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).
ET1606-55_5.jpg Fig5: ICC staining of Notch 1 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 (ET1606-55, 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).
ET1606-55_6.jpg Fig6: Immunohistochemical analysis of paraffin-embedded human pancreas tissue using anti-Notch 1 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) 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 (ET1606-55, 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.
ET1606-55_7.jpg Fig7: Immunohistochemical analysis of paraffin-embedded mouse lung tissue using anti-Notch 1 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) 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 (ET1606-55, 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.
ET1606-55_8.jpg Fig8: Immunohistochemical analysis of paraffin-embedded mouse pancreas tissue using anti-Notch 1 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) 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 (ET1606-55, 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.
ET1606-55_9.jpg Fig9: Immunofluorescence analysis of paraffin-embedded human liver cancer tissue labeling Notch 1 with Rabbit anti-Notch 1 antibody (ET1606-55) at 1/50 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 10% negative goat serum for 1 hour at room temperature, washed with PBS, and then probed with the primary antibody (ET1606-55, green) at 1/50 dilution overnight at 4 ℃, washed with PBS. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. Nuclei were counterstained with DAPI (blue).
ET1606-55_10.jpg Fig10: Flow cytometric analysis of Notch 1 was done on Hela cells. The cells were fixed, permeabilized and stained with the primary antibody (ET1606-55, 1/50) (blue). 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; red).
ET1606-55_11.jpg Fig11: Western blot analysis of Notch 1 on different lysates with Rabbit anti-Notch 1 antibody (ET1606-55) at 1/2,000 dilution.

Lane 1: C6 cell lysate
Lane 2: Rat brain tissue lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 272 kDa
Observed band size: 125 kDa

Exposure time: 60 seconds;

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 (ET1606-55) 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.
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.