PRODUCT CODE: ER1902-75

Ki67 Antibody (ER1902-75)

Applications

  • WB

  • IHC-P

  • ICC

REACTIVITY

  • Human

Western blot analysis of Ki67 on HepG2 cell lysate. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ER1902-75, 1/10000) 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.
  • Western blot analysis of Ki67 on HepG2 cell lysate. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ER1902-75, 1/10000) 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.
  • ICC staining of Ki67 in A549 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 (ER1902-75, 1/200) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/100 dilution. The nuclear counter stain is DAPI (blue).
  • Immunohistochemical analysis of paraffin-embedded human colon cancer 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 (ER1902-75, 1/400) 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.
Western blot analysis of Ki67 on HepG2 cell lysate. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ER1902-75, 1/10000) 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.

Applications

  • WB

  • IHC-P

  • ICC

REACTIVITY

  • Human

SPECIFICATIONS

Product Type

Rabbit polyclonal primary

Product Name

Ki67 Antibody (ER1902-75)

Immunogen

Synthetic peptide within human ki67 aa 1900-2000.

Host

Rabbit

Positive Control

HepG2 cell lysates, A549, human colon cancer tissue.

Conjugation

Unconjugated

Clonality

Polyclonal

PROPERTIES

Form

Liquid

Storage Condition

Store at +4C after thawing. Aliquot store at -20C. Avoid repeated freeze / thaw cycles.

Storage Buffer

1*PBS (pH7.4), 0.2% BSA, 50% Glycerol. Preservative: 0.05% Sodium Azide.

Concentration

1 ug/ul

PURIFICATION

Peptide affinity purified.

MOLECULAR WEIGHT

Predicted band size 359 kDa.

Isotype

IgG

APPLICATION DILUTION

  • WB

  • 1:10,000-1:50,000

  • IHC-P

  • 1:200-1:1,000

  • ICC

  • 1:50-1:200

TARGET

UNIPROT #

PROTEIN NAME

Ki67

SYNONYMS

Antigen identified by monoclonal antibody Ki 67 antibody; Antigen identified by monoclonal antibody Ki-67 antibody; Antigen KI-67 antibody; Antigen KI67 antibody; Antigen Ki67 antibody; KI67_HUMAN antibody; KIA antibody; Marker of proliferation Ki-67 antibody; MIB 1 antibody; MIB antibody; MKI67 antibody; PPP1R105 antibody; Proliferation marker protein Ki-67 antibody; Proliferation related Ki 67 antigen antibody; Protein phosphatase 1 regulatory subunit 105 antibody; RP11-380J17.2 antibody

DEVELOPMENTAL STAGE

Expression occurs preferentially during late G1, S, G2 and M phases of the cell cycle, while in cells in G0 phase the antigen cannot be detected (at protein level). Present at highest level in G2 phase and during mitosis (at protein level). In interphase, forms fiber-like structures in fibrillarin-deficient regions surrounding nucleoli.

POST-TRANSLATIONAL MODIFICATION

Phosphorylated. Hyperphosphorylated in mitosis. Hyperphosphorylated form does not bind DNA.

SUBCELLULAR LOCATION

Chromosome, Nucleus

FUNCTION

Required to maintain individual mitotic chromosomes dispersed in the cytoplasm following nuclear envelope disassembly. Associates with the surface of the mitotic chromosome, the perichromosomal layer, and covers a substantial fraction of the chromosome surface. Prevents chromosomes from collapsing into a single chromatin mass by forming a steric and electrostatic charge barrier: the protein has a high net electrical charge and acts as a surfactant, dispersing chromosomes and enabling independent chromosome motility. Binds DNA, with a preference for supercoiled DNA and AT-rich DNA. Does not contribute to the internal structure of mitotic chromosomes. May play a role in chromatin organization. It is however unclear whether it plays a direct role in chromatin organization or whether it is an indirect consequence of its function in maintaining mitotic chromosomes dispersed (Probable).