CD99 Recombinant Rabbit Monoclonal Antibody [JF0991]
cat.: ET1702-35
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human
Applications: WB, IF-Cell, IF-Tissue, IHC-P, IP, FC
Clonality: Monoclonal
Clone number: JF0991
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: 25 kDa
Isotype: IgG
Immunogen: Recombinant protein within Human CD99 aa 1-130 / 185.
Positive control: THP-1 cell lysates, Hela, PANC-1, PC-3M, human pancreas tissue, human uterus tissue, human tonsil tissue, Jurkat.
Subcellular location: Membrane.
Recommended Dilutions:
  WB
  IF-Cell
  IF-Tissue
  IHC-P
  FC
1:1,000-1:2,000
1:50-1:200
1:50-1:200
1:50-1:200
1:50-1:100
Uniprot #: SwissProt: P14209 Human
Alternative names: 12E7 Antigen identified by monoclonal 12E7, Y homolog Antigen identified by monoclonal antibodies 12E7, F21 and O13 CD99 CD99 antigen CD99 molecule CD99_HUMAN Cell surface antigen 12E7 Cell surface antigen HBA 71 Cell surface antigen O13 E2 antigen HBA71 MIC 2X MIC 2Y MIC2 (monoclonal 12E7) MIC2 MIC2X MIC2Y MSK5X Protein MIC2 Surface antigen MIC2 T cell surface glycoprotein E2 T-cell surface glycoprotein E2
Images
ET1702-35_1.jpg Fig1: Western blot analysis of CD99 on THP-1 cell lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1702-35, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:40,000 dilution was used for 1 hour at room temperature.
ET1702-35_2.jpg Fig2: ICC staining of CD99 in Hela cells (red). 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 (ET1702-35, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®594 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
ET1702-35_3.jpg Fig3: ICC staining of CD99 in PANC-1 cells (red). 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 (ET1702-35, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®594 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
ET1702-35_4.jpg Fig4: ICC staining of CD99 in PC-3M cells (red). 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 (ET1702-35, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®594 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
ET1702-35_5.jpg Fig5: Immunohistochemical analysis of paraffin-embedded human pancreas tissue using anti-CD99 antibody. 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 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1702-35, 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.
ET1702-35_6.jpg Fig6: Immunohistochemical analysis of paraffin-embedded human uterus tissue using anti-CD99 antibody. 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 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1702-35, 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.
ET1702-35_7.jpg Fig7: Immunohistochemical analysis of paraffin-embedded human tonsil tissue using anti-CD99 antibody. 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 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1702-35, 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.
ET1702-35_8.jpg Fig8: Flow cytometric analysis of CD99 was done on Jurkat cells. The cells were fixed, permeabilized and stained with the primary antibody (ET1702-35, 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”.