Ki67 Recombinant Rabbit Monoclonal Antibody [ST50-01]
cat.: ET1609-34
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human
Applications: WB, IF-Cell, IHC-P, FC
Clonality: Monoclonal
Clone number: ST50-01
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: 359 kDa
Isotype: IgG
Immunogen: Synthetic peptide within human Ki67 aa 1040-1080.
Positive control: HepG2 cell lysates, HepG2, HeLa, human tonsil tissue,human stomach carcinoma tissue, human colon carcinoma tissue.
Subcellular location: Nucleus, Chromosome.
Recommended Dilutions:
  WB
  IF-Cell
  IHC-P
  FC
1:1,000-1:2,000
1:50-1:1,000
1:100-1:500
1:50-1:100
Uniprot #: SwissProt: P46013 Human
Alternative names: Antigen identified by monoclonal Ki 67 Antigen identified by monoclonal Ki-67 Antigen KI-67 Antigen KI67 Antigen Ki67 KI67_HUMAN KIA Marker of proliferation Ki-67 MIB 1 MIB MKI67 PPP1R105 Proliferation marker protein Ki-67 Proliferation related Ki 67 antigen Protein phosphatase 1 regulatory subunit 105 RP11-380J17.2
Images
ET1609-34_1.jpg Fig1: Western blot analysis of Ki67 on HepG2 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 (ET1609-34, 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.
ET1609-34_2.jpg Fig2: ICC staining of Ki67 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 (ET1609-34, 1/1,000) 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).
ET1609-34_3.jpg Fig3: Immunocytochemistry analysis of HeLa cells labeling Ki67 with Rabbit anti-Ki67 antibody (ET1609-34) 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-Ki67 antibody (ET1609-34) 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.
ET1609-34_4.jpg Fig4: Immunohistochemical analysis of paraffin-embedded human stomach carcinoma tissue using anti-Ki67 antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) 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 (ET1609-34, 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.
ET1609-34_5.jpg Fig5: Immunohistochemical analysis of paraffin-embedded human colon carcinoma tissue using anti-Ki67 antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) 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 (ET1609-34, 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.
ET1609-34_6.jpg Fig6: Immunohistochemical analysis of paraffin-embedded human tonsil tissue with Rabbit anti-Ki67 antibody (ET1609-34) at 1/800 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 (ET1609-34) at 1/800 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.
ET1609-34_7.jpg Fig7: Flow cytometric analysis of Ki67 was done on Hela cells. The cells were fixed, permeabilized and stained with the primary antibody (ET1609-34, 1/100) (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).
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.