SESN1 Recombinant Rabbit Monoclonal Antibody [JE36-66]
cat.: HA722307
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human
Applications: WB, IF-Cell, IHC-P, FC
Clonality: Monoclonal
Clone number: JE36-66
Form: Liquid
Storage condition: Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term.
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: 57 kDa
Isotype: IgG
Immunogen: Synthetic peptide within Human SESN1 aa 100-200.
Positive control: HeLa cell lysate, K-562 cell lysate, Ramos cell lysate, K-562, human breast cancer tissue, mouse heart tissue.
Subcellular location: Nucleus, Cytoplasm.
Recommended Dilutions:
  WB
  IF-Cell
  IHC-P
  FC

1:1,000
1:100
1:1,000
1:1,000
Uniprot #: SwissProt: Q9Y6P5 Human
Alternative names: 1110002G11Rik AU044290 MGC118148 MGC138241 MGC142129 OTTMUSP00000022799 p53 activated gene 26 p53 regulated PA26 nuclear protein p53 regulated protein PA26 p53-regulated protein PA26 PA26 RP11-787I22.1 sesn1 SESN1_HUMAN SEST1 sestrin 1 Sestrin-1
Images
HA722307_1.jpg Fig1: Western blot analysis of SESN1 on different lysates with Rabbit anti-SESN1 antibody (HA722307) at 1/1,000 dilution.

Lane 1: HeLa cell lysate
Lane 2: K-562 cell lysate
Lane 3: Ramos cell lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 57 kDa
Observed band size: 70 kDa

Exposure time: Lane 1-2: 24 seconds; Lane 3: 1 minute 2 seconds; K1802;

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 (HA722307) 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.
HA722307_2.jpg Fig2: Immunocytochemistry analysis of K-562 cells labeling SESN1 with Rabbit anti-SESN1 antibody (HA722307) 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-SESN1 antibody (HA722307) 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.
HA722307_3.jpg Fig3: Immunohistochemical analysis of paraffin-embedded human breast cancer tissue with Rabbit anti-SESN1 antibody (HA722307) 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 (HA722307) 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.
HA722307_4.jpg Fig4: Immunohistochemical analysis of paraffin-embedded mouse heart tissue with Rabbit anti-SESN1 antibody (HA722307) 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 (HA722307) 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.
HA722307_5.jpg Fig5: Flow cytometric analysis of K-562 cells labeling SESN1.

Cells were fixed and permeabilized. Then stained with the primary antibody (HA722307, 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).
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.