Ki-67 antibody (AA 1547-1742)

Catalog No. ABIN968284

Product Details anti-Ki-67 Antibody

Target: Ki-67 (MKI67) (Antigen Identified By Monoclonal Antibody Ki-67 (MKI67))

Binding Specificity: AA 1547-1742

Reactivity: Human

Host: Mouse

Clonality: Monoclonal

Conjugate: This Ki-67 antibody is un-conjugated

Application: Western Blotting (WB), BioImaging (BI)

Characteristics: 1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
2. Please refer to us for technical protocols.
3. This antibody has been developed and certified for the bioimaging application. However, a routine bioimaging test is not performed on every lot. Researchers are encouraged to titrate the reagent for optimal performance.
4. Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
5. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
6. Triton is a trademark of the Dow Chemical Company.

Purification: The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography.

Immunogen: Human Ki-67 aa. 1547-1742

Clone: 35-Ki

Isotype: IgG1

Application Details

Application Notes: Bioimaging
1. Seed the cells in appropriate culture medium at ~10,000 cells per well in an 96-well Imaging Plate and culture overnight.
2. Remove the culture medium from the wells, and fix the cells by adding 100 myl of Fixation Buffer to each well. Incubate for 10 minutes at room temperature (RT).
3. Remove the fixative from the wells, and permeabilize the cells using either 90% methanol, or Triton™ X-100: a. Add 100 myl of -20°C 90% methanol to each well and incubate for 5 minutes at RT. OR b. Add 100 myl of 0.1% Triton™ X-100 to each well and incubate for 5 minutes at RT.
4. Remove the permeabilization buffer, and wash the wells twice with 100 myl of 1× PBS.
5. Remove the PBS, and block the cells by adding 100 myl of to each well. Incubate for 30 minutes at RT.
6. Remove the blocking buffer and add 50 myl of the optimally titrated primary antibody (diluted in Stain Buffer) to each well, and incubate for 1 hour at RT.
7. Remove the primary antibody, and wash the wells three times with 100 myl of 1× PBS.
8. Remove the PBS, and add the second step reagent at its optimally titrated concentration in 50 myl to each well, and incubate in the dark for 1 hour at RT.
9. Remove the second step reagent, and wash the wells three times with 100 myl of 1× PBS.
10. Remove the PBS, and counter-stain the nuclei by adding 200 myl per well of 2 myg/ml Hoechst 33342 in 1× PBS to each well at least 15 minutes before imaging.
11. View and analyze the cells on an appropriate imaging instrument.

Comment:

Related Products: ABIN967389, ABIN968535

Restrictions: For Research Use only

Handling

Format: Liquid

Concentration: 250 μg/mL

Buffer: Aqueous buffered solution containing BSA, glycerol, and ≤0.09 % sodium azide.

Preservative: Sodium azide

Precaution of Use: This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.

Storage: -20 °C

Storage Comment: Store undiluted at -20°C.

References for anti-Ki-67 antibody (ABIN968284)

Saitoh, Pizzi, Wang: "Perturbation of SUMOlation enzyme Ubc9 by distinct domain within nucleoporin RanBP2/Nup358." in: The Journal of biological chemistry, Vol. 277, Issue 7, pp. 4755-63, (2002) (PubMed).

Starborg, Gell, Brundell, Höög: "The murine Ki-67 cell proliferation antigen accumulates in the nucleolar and heterochromatic regions of interphase cells and at the periphery of the mitotic chromosomes in a process essential for cell cycle progression." in: Journal of cell science, Vol. 109 ( Pt 1), Issue 12, pp. 143-53, (1996) (PubMed).

Schlüter, Duchrow, Wohlenberg, Becker, Key, Flad, Gerdes: "The cell proliferation-associated antigen of antibody Ki-67: a very large, ubiquitous nuclear protein with numerous repeated elements, representing a new kind of cell cycle-maintaining proteins." in: The Journal of cell biology, Vol. 123, Issue 3, pp. 513-22, (1993) (PubMed).

Target Details for Ki-67

Target: Ki-67 (MKI67) (Antigen Identified By Monoclonal Antibody Ki-67 (MKI67))

Alternative Name: Ki-67 (MKI67 Products)

Synonyms: MKI67 antibody, kia antibody, mki67 antibody, MGC132156 antibody, D630048A14Rik antibody, Ki-67 antibody, Ki67 antibody, KIA antibody, MIB-1 antibody, marker of proliferation Ki-67 antibody, marker of proliferation Ki-67 L homeolog antibody, marker of proliferation Ki-67 S homeolog antibody, antigen identified by monoclonal antibody Ki 67 antibody, antigen identified by monoclonal antibody Ki-67 antibody, nucleolar protein interacting with the FHA domain of MKI67 antibody, MKI67 antibody, mki67 antibody, mki67.L antibody, mki67.S antibody, Mki67 antibody, NIFK antibody

Background: Cellular proliferation is a complex multi-faceted process, central to biological events ranging from embryonic development to wound healing. Although proliferation is tightly controlled, dysregulation often results in tumorigenesis. The regulatory process involves the expression of many cell-cycle-associated proteins that are subject to cycle-dependent modification. Such protein expression may be monitored to assess the proliferative capacity of cells, especially tumorigenic cells.The Ki-67 antigen is a nuclear protein expressed exclusively in proliferating cells during all active parts of the cell cycle. However, it is absent in quiescent cells and during DNA repair. The distribution of Ki-67 changes during different stages of the cell cycle. During G1, it is localized to the perinuclear region, but is primarily found in the nuclear matrix in later phases. During mitosis, it associates with the condensed chromosomes. The nuclear localization of Ki-67 and strict association with the cell cycle indicate its importance in the regulation of cell division. Therefore, Ki-67 has become an important marker of proliferating cells, and may also be a marker for disitinct nuclear matrix compartments.

Molecular Weight: 395 kDa

Pathways: Glycosaminoglycan Metabolic Process