Antigen Identified By Monoclonal Antibody Ki-67 (MKI67) (C-Term) antibody

Details for Product No. ABIN269440
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Antigen
Synonyms MKI67, kia, mki67, MGC132156, D630048A14Rik, Ki-67, Ki67, KIA, MIB-1
Epitope
C-Term
(37), (7), (6), (5), (2), (2), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1)
Reactivity
Human
(199), (30), (28), (2), (2), (1), (1)
Host
Rabbit
(125), (74), (6), (2), (2)
Clonality (Clone)
Monoclonal ()
Conjugate
Un-conjugated
(2), (2), (2), (1), (1), (1), (1), (1), (1), (1), (1)
Application
Immunocytochemistry (ICC), Immunofluorescence (IF), Immunohistochemistry (IHC), Immunohistochemistry (Frozen Sections) (IHC (fro)), Immunohistochemistry (Paraffin-embedded Sections) (IHC (p))
(98), (70), (64), (58), (35), (29), (19), (10), (10), (1), (1)
Pubmed 1 reference available
Quantity 0.5 mL
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Catalog No. ABIN269440
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Immunogen A synthetic peptide from C-terminus of human Ki-67.
Clone SP6
Isotype IgG
Specificity Ki67 (SP6)
Cross-Reactivity (Details) Mouse reactivity reported in scientific literature (PMID: 22020958).
Purification Tissue culture supernatant
Alternative Name Ki-67
Background Ki67 antigen is the prototypic cell cycle related nuclear protein, expressed byproliferating cells in all phases of the active cell cycle (G1, S, G2 and M phase). It isabsent in resting (G0) cells. Ki67 antibodies are useful in establishing the cell growingfraction in neoplasms (immunohistochemically quantified by determining the number ofKi67 positive cells among the total number of resting cells = Ki67 index). In neoplastictissues the prognostic value is comparable to the tritiated thymidine labelling index. Thecorrelation between low Ki67 index and histologically low grade tumours is strong. Ki67is routinely used as a neuronal marker of cell cycling and proliferation. Alternate Names: anti-Antigen identified by monoclonal antibody Ki 67 antibody, anti-Antigen KI67antibody, anti-KIA antibody, anti-MKI67 antibody, anti-Proliferation related Ki 67 antigenantibody.
Gene Symbol: MKI67
Gene ID 4288, 291234
Research Area Proliferation Markers, Cell Cycle
Application Notes Suggested incubation period of 30 minutes at room temperature. However, depending upon the fixation conditions and the staining system employed, optimal incubation should be determined by the user. Formalin fixed paraffin embedded tissue sections require high temperature antigen unmasking with 10 mM citrate buffer, pH 6.0 prior to immunostaining. Optimal dilutions/concentrations should be determined by the end user. Use in Immunocytochemistry/immunofluorescence reported in scientific literature (PMID 20235278) Use in Immunohistochemistry-Frozen reported in scientific literature (PMID 23300752)
Recommended dilutions: Immunocytochemistry/Immunofluorescence 1:10-1:500, Immunohistochemistry 1:10-1:500, Immunohistochemistry-Frozen 1:10-1:500, Immunohistochemistry-Paraffin 1:100-1:200
Protocol Immunohistochemistry-Paraffin Protocol Specific for NB600-1252: Ki67 Antibody (SP6)Materials1) 1 Phosphate buffered saline (pH 7.6): NaCl 137mmol/L, KCl 2.7mmol/L, Na2HPO4 4.3mmol/L, KH2PO4 1.4 mmol/L2) Citrate buffer, 0.01 M, pH6.0, Sodium Citrate 3g, Citric acid 0.4g3) 3 % Hydrogen peroxide4) Primary antibody5) Blocking serum (normal serum)6) Biotinylated secondary antibody7) DAB staining kitMethods
1. Dewax and hydration of slides using xylene and EtOH:Dry slides for 20 min in a 60 C ovenAdd Xylene, 2 x 10 min100 %, 95 %, 80 %, and 70 % EtOH, 5 min each EtOH concentration Rinse in PBS, 5'2 Antigen retrieval method (only for paraffin slides)1a. High-pressure antigen retrieval procedure (recommended method)Place slides in a glass slide holder (ensure that the slide holder is completely filled with slides, slides without sections if necessary, to ensure even heating. The entire slide holder is immersed in 1000 mL of Citrate buffer (0.01M, pH6.
0) within a pressure cookerOnce steam is produced, and ONLY when steam is visible, from the pressure cooker (usually 15-20 min), the required high-pressure will have been reached, and slides will be incubated for 2 min.Turn off heat, and allow buffer and slides to cool to room temperatureSlides are then rinsed in PBS for 5 minutes
. Add 3 % hydrogen peroxide solution, 10'at RT, then PBS, 3X5'
. Normal blocking serum, 20'at RT
. Incubate with Primary Ab, 4C overnight or 1.5 hours at 37C
. Rinse with PBS, 3 X 5' each rinse
. Add Biotin-conjugated second antibody, 10'at RT
. Rinse with PBS, 3 X 5' each rinse
. Add Streptavidin-Peroxidase, 10'at RT
. Rinse with PBS, 3 X 5' each rinse
. Staining with DAB solution, 2-5'under microscope
. Stop the reaction by washing in tap water
. Counterstain in Haematoxylin for 3-5 minutes
. 75 %, 80 %, 95 % and 100 % ethanol, 5x2', xylene 2 x 10'
Restrictions For Research Use only
Format Liquid
Buffer PBS, BSA, Sodium Azide
Preservative Sodium azide
Precaution of Use WARNING: Reagents contain sodium azide. Sodium azide is very toxic if ingested or inhaled. Avoid contact with skin, eyes, or clothing. Wear eye or face protection when handling. If skin or eye contact occurs, wash with copious amounts of water. If ingested or inhaled, contact a physician immediately. Sodium azide yields toxic hydrazoic acid under acidic conditions. Dilute azide-containing compounds in running water before discarding to avoid accumulation of potentially explosive deposits in lead or copper plumbing.
Handling Advice Do not freeze.
Storage 4 °C
General Koya, Lu, Sun et al.: "Reversal of streptozotocin-induced diabetes in mice by cellular transduction with recombinant pancreatic transcription factor pancreatic duodenal homeobox-1: a novel protein transduction domain-based therapy." in: Diabetes, Vol. 57, Issue 3, pp. 757-69, 2008 (PubMed).

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