Fragilis Recombinant Rabbit Monoclonal Antibody [JU73-02]
cat.: ET1706-09
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human
Applications: WB, IF-Cell, IF-Tissue, IHC-P, FC
Clonality: Monoclonal
Clone number: JU73-02
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: 15 kDa
Isotype: IgG
Immunogen: Synthetic peptide within Human Fragilis aa 1-50 / 133.
Positive control: HepG2 cell lysate, HeLa cell lysate, THP-1 cell lysate, HeLa, human breast cancer tissue, human colon cancer tissue, MCF-7.
Subcellular location: Cell membrane, Late endosome membrane, Lysosome membrane, perinuclear region.
Recommended Dilutions:
  WB
  IF-Cell
  IF-Tissue
  IHC-P
  FC
1:2,000
1:100
1:50-1:200
1:1,000
1:50-1:100
Uniprot #: SwissProt: Q01628 Human
Alternative names: 1 8U Fragilis IFITM3 IFM3_HUMAN Interferon induced transmembrane protein 3 (1 8U) Interferon Induced Transmembrane Protein 3 Interferon inducible Interferon inducible protein 1 8U Interferon Inducible Protein 15 Interferon Inducible Protein Homolog Interferon-induced transmembrane protein 3 Interferon-inducible protein 1-8U IP15
Images
ET1706-09_1.jpg Fig1: Western blot analysis of Fragilis on different lysates with Rabbit anti-Fragilis antibody (ET1706-09) at 1/2,000 dilution.

Lane 1: HepG2 cell lysate
Lane 2: HeLa cell lysate
Lane 3: THP-1 cell lysate
Lane 4: 293T cell lysate (negative)

Lysates/proteins at 15 µg/Lane.

Predicted band size: 15 kDa
Observed band size: 15 kDa

Exposure time: 4 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 (ET1706-09) 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.
ET1706-09_2.jpg Fig2: Immunocytochemistry analysis of HeLa cells labeling Fragilis with Rabbit anti-Fragilis antibody (ET1706-09) at 1/100 dilution.

Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 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-Fragilis antibody (ET1706-09) 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.
ET1706-09_3.jpg Fig3: Immunohistochemical analysis of paraffin-embedded human breast cancer tissue with Rabbit anti-Fragilis antibody (ET1706-09) at 1/200 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 (ET1706-09) 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.
ET1706-09_4.jpg Fig4: Immunohistochemical analysis of paraffin-embedded human colon cancer tissue with Rabbit anti-Fragilis antibody (ET1706-09) at 1/200 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 (ET1706-09) 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.
ET1706-09_5.jpg Fig5: Immunofluorescence analysis of paraffin-embedded human breast cancer tissue labeling Fragilis with Rabbit anti-Fragilis antibody (ET1706-09) at 1/200 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 10% negative goat serum for 1 hour at room temperature, washed with PBS, and then probed with the primary antibody (ET1706-09, green) at 1/200 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).
ET1706-09_6.jpg Fig6: Flow cytometric analysis of Fragilis was done on MCF-7 cells. The cells were fixed, permeabilized and stained with the primary antibody (ET1706-09, 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).
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.