CAF-1 Subunit antibody

Details for Product No. ABIN153017
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Antigen
Reactivity
Fruit Fly (Drosophila melanogaster), Human, Yeast
(5), (1)
Host
Mouse
(7)
Clonality (Clone)
Monoclonal ()
Application
Western Blotting (WB), Immunocytochemistry (ICC), Immunofluorescence (IF), Immunoprecipitation (IP)
(6), (5), (4), (2), (1), (1), (1)
Pubmed 2 references available
Quantity 0.05 mL
Shipping to United States (Change)
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Catalog No. ABIN153017
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Immunogen Recombinant human CHAF1B
Clone SS 24 1-68
Isotype IgG1
Purification Ascites
Background Chromatin assembly factor 1 (CAF-1) is a member of the histone chaperone family. It isrequired for inheritance of epigenetically determined chromosomal states, and in vitro, itpromotes the assembly of chromatin during DNA replication and DNA repair. CAF-1 isa complex of three subunits, p150, p60, and p48. This complex is localized to thenucleus of human cells.Both the composition and the biochemical activity of CAF-1 are conserved amonghuman, yeast, and Drosophila. Alternate Names: anti-p60 CAF1 antibody, CHAF1B antibody Chromatin assembly factor 1 subunit B antibody, anti-Chromatin assembly factor Iantibody, anti-M phase phosphoprotein 7 antibody, anti-MPHOSPH7 antibody.
Gene Symbol: CHAF1B
Gene ID 8208
UniProt Q13112
Application Notes This CHAF1B (SS 24 1-68) antibody is useful for Immunoprecipitation, Western blot and Immunocytochemistry/Immunofluorescence. In WB a band can be seen at ~60 kDa.
Recommended dilutions: Immunocytochemistry/Immunofluorescence 1:10-1:2000, Immunoprecipitation 0.1-1 µg, Western Blot 1:1000
Protocol Immunoprecipitation protocol specific for CAF-1 p60 Antibody Procedure Guide for Monoclonal anti- CAF-1 p60 (clone 24)Catalog# NB 500-211Immunoprecipitation Procedure1) Cells were prepared in 0.4-1 % NP-40 lysis buffer.
2) After osmotic shock and nuclear extractions of the cells, use the following buffer: 25mM Tris, 150mM NaCl, 1mM EDTA, 1mM DTT, and 0.01 % NP-
40. 3) Clarify the extracts with two successive 5 minute clearing spins (in a microfuge).
) Preclear the extracts with some protein G beads (~20 µL). 5) Incubate the extracts with ~ 0.1-1ug of anti-CAF-1p60-24 [NB 500-211]/ control antibody for 1-3 hours. 6) Spin the antibody/extract for 5 minutes.
) Incubate the antibody/extract with protein G beads, rotating for 1-2 hours.
) Wash the beads, twice, for 5 minutes, at low ionic strength [150mM MaCl] in 1ml volume (*Optional: wash again, twice, at high ionic strength [ 400mM NaCl]).
) Resuspend the washed beads in Laemlli sample buffer and boil for 5 minutes. NOTE: a. All steps for IP are performed at 4C.b. CAF-1p60 is phosphorylated and contains a PEST sequence. Therefore, it is recommended that protease and phophatase inhibitors be added to all buffers.c. The protocol for crosslinking CAF-1p60 to protein G beads was obtained from the Harlow and Lane book, Using Antibodies.
Restrictions For Research Use only
Format Liquid
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 Avoid freeze-thaw cycles
Storage -20 °C
Storage Comment Aliquot and store at -20 °C or -80 °C.
General Smith, Stillman: "Immunological characterization of chromatin assembly factor I, a human cell factor required for chromatin assembly during DNA replication in vitro." in: The Journal of biological chemistry, Vol. 266, Issue 18, pp. 12041-7, 1991 (PubMed).

Shibahara, Stillman: "Replication-dependent marking of DNA by PCNA facilitates CAF-1-coupled inheritance of chromatin." in: Cell, Vol. 96, Issue 4, pp. 575-85, 1999 (PubMed).

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