Human Ki67 Recombinant Rabbit Monoclonal Antibody [SR00-02]
cat.: HA724537
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human
Applications: ELISA(Det)
Clonality: Monoclonal
Clone number: SR00-02
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
Storage buffer: 1*PBS (pH7.4).
Concentration: 1ug/ul
Purification: Protein A affinity purified.
Isotype: IgG
Immunogen: Recombinant protein within Human Ki67 aa 1,000-1,234 (HA211578).
Positive control: Recombinant Human Ki67 protein (HA211578).
Subcellular location: Chromosome, Nucleus, nucleolus.
Recommended Dilutions:
  ELISA(Det)

Use at an assay dependent concentration. Can be paired for Sandwich ELISA with Rabbit monoclonal [PSH25-46] to Human Ki67 (Capture) (HA724536) and Recombinant Human Ki67 protein (HA211578) as the standard. The reference range value is 31.3-8,000 pg/mL.
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
HA724537_1.png Fig1: Sandwich ELISA analysis of Human Ki67 matched pair antibodies

Capture: HA724536, Human Ki67 Rabbit mAb [PSH25-46]
Detector: HA724537, Human Ki67 Rabbit mAb [SR00-02]

Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA724536) diluted in carbonate/bicarbonate buffer, at a concentration of 5 μg/mL overnight at 4℃. Wells of the plate were washed, blocked with 150 µl 0.05% tween-20 1% BSA blocking buffer, and incubated with serial diluted Recombinant Human Ki67 protein (HA211578) starting from 8,000 pg/mL to 0 pg/mL and detect antibody (HA724537, Biotin, 0.2 μg/mL) for 1 hour at 30℃ with shaking. Then the plate was washed and incubated with 100 µl per well of SA-HRP for 0.5 hour at 30℃ with shaking. Detection was performed using an Ultra TMB Substrate for 10 minutes at room temperature in the dark. The reaction was stopped with sulfuric acid and absorbances were read on a spectrophotometer at 450 nm.
HA724537_2.png Fig2: Interpolated concentrations of native Ki67 in A431 and HeLa cell extract samples based on a 1,000 µg/mL extract load.

Capture: HA724536, Human Ki67 Rabbit mAb [PSH25-46]
Detector: HA724537, Human Ki67 Rabbit mAb [SR00-02]

The concentrations of Ki67 were measured in duplicates, interpolated from the Ki67 standard curve and corrected for sample dilution. Undiluted samples are A431 cell extract 50% and HeLa cell extract 25%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). The mean Ki67 concentration was determined to be 13,077 pg/mL in A431 cell extract and 39,701 pg/mL in HeLa cell extract.
HA724537_3.png Fig3: Interpolated concentrations of spiked Ki67 in human cell culture media samples.

Capture: HA724536, Human Ki67 Rabbit mAb [PSH25-46]
Detector: HA724537, Human Ki67 Rabbit mAb [SR00-02]

The concentrations of Ki67 were measured in duplicates, interpolated from the Ki67 standard curves and corrected for sample dilution. Diluted samples are as follows: 50% cell culture media with FBS. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2).
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.