CDC5 Cell Division Cycle 5-Like (S. Pombe) (CDC5L) (AA 109-303) antibody

Details for Product No. ABIN968837
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Antigen
Synonyms
zgc:55853, CDC5L, CDC5, CDC5-LIKE, CEF1, PCDC5RP, dJ319D22.1, 1200002I02Rik, AA408004, ARABIDOPSIS THALIANA CELL DIVISION CYCLE 5, ARABIDOPSIS THALIANA MYB DOMAIN CELL DIVISION CYCLE 5, ATCDC5, ATMYBC ... show more
zgc:55853, CDC5L, CDC5, CDC5-LIKE, CEF1, PCDC5RP, dJ319D22.1, 1200002I02Rik, AA408004, ARABIDOPSIS THALIANA CELL DIVISION CYCLE 5, ARABIDOPSIS THALIANA MYB DOMAIN CELL DIVISION CYCLE 5, ATCDC5, ATMYBCDC5, F21M12.15, F21M12_15, cell division cycle 5 show less
Epitope
AA 109-303
(7), (5), (2), (2), (2), (1), (1), (1), (1), (1), (1)
Reactivity
Rat (Rattus)
(46), (22), (21), (13), (12), (1), (1), (1), (1)
Host
Mouse
(39), (7)
Clonality (Clone)
Monoclonal ()
Conjugate
Un-conjugated
(2), (2), (2), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1)
Application
Western Blotting (WB), BioImaging (BI)
(30), (17), (15), (12), (10), (5), (5), (4), (3), (2), (1)
Pubmed 4 references available
Quantity 50 μg
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Catalog No. ABIN968837
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Immunogen Rat CDC5L
Clone LPR-01
Isotype IgG1
Cross-Reactivity Chicken, Dog (Canine), Human, Mouse (Murine)
Characteristics 1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
2. 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.
3. Triton is a trademark of the Dow Chemical Company.
4. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
5. 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.
6. Please refer to us for technical protocols.
Purification Purified from tissue culture supernatant or ascites by affinity chromatography.
Alternative Name CDC5L
Background Splicing, the removal of introns from pre-mRNA, is mediated by spliceosomal complexes and occurs in two distinct catalytic steps. The first step involves cleavage of the 5' exon and the production of a lariat intermediate. In the second step, the 3'-splice site is cleaved and the exons are fused with concomitant release of the intron lariat. The spliceosome contains multiple snRNPs and a number of non-snRNP splicing factors. CDC5L is a non-snRNP component of the splicesome that is homologous to the yeast Cdc5 protein. The sequence for CDC5L contgains a helix-turn-helix DNA binding domain (DBD), four nuclear localization signals, and a hydrophilic, proline-rich central region that is similar to the transcriptional activating domain in Myb family transcription factors. CDC5L can interact with the nuclear PP1 inhibitor NIPP-1 and the WD40 domain protein PLRG1. Both of these interactions are critical for pre-mRNA splicing and may also be important for G 2/M phase transition. In addition, CDC5L translocates from the cytoplasm to the nucleus in the presence of serum. Thus, CDC5L localization and splicesome-regulating activity may be controlled by mitogen-activated signal transduction pathways.
Molecular Weight 105 kDa
Research Area Chromatin and Nuclear Signaling, DNA/RNA
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: ABIN968546, ABIN967389

Restrictions For Research Use only
Format Liquid
Concentration 250 µg/ml
Buffer Aqueous buffered solution containing BSA, glycerol.
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
Supplier Images
anti-CDC5 Cell Division Cycle 5-Like (S. Pombe) (CDC5L) (AA 109-303) antibody Immunofluorescent staining of U-2 OS (ATCC HTB-96) cells. Cells were seeded in a 96 well imaging plate at ~ 10 000 cells per well. After overnight incubation, cells were stained using the alcohol perm protocol and the anti-CDC5L antibody. The second step reagent was FITC goat anti mouse Ig. Images were taken on a BD Pathway™ 855 bioimaging system using a 20x objective. This antibody also stained HeLa (ATCC CCL-2) cells and worked with both the Triton™ X-100 and alcohol perm protocols. This antibody is not recommended for staining A549 cells.
anti-CDC5 Cell Division Cycle 5-Like (S. Pombe) (CDC5L) (AA 109-303) antibody (2) Western blot analysis of CDC5L on a rat cerebrum lysate. Lane 1: 1:5000, lane 2: 1:10000, lane 3: 1:20000 dilution of the CDC5L antibody.
Product cited in: Bernstein, Coughlin: "Pombe Cdc5-related protein. A putative human transcription factor implicated in mitogen-activated signaling." in: The Journal of biological chemistry, Vol. 272, Issue 9, pp. 5833-7, 1997 (PubMed).

Boudrez, Beullens, Groenen et al.: "NIPP1-mediated interaction of protein phosphatase-1 with CDC5L, a regulator of pre-mRNA splicing and mitotic entry." in: The Journal of biological chemistry, Vol. 275, Issue 33, pp. 25411-7, 2000 (PubMed).

Ajuh, Kuster, Panov et al.: "Functional analysis of the human CDC5L complex and identification of its components by mass spectrometry." in: The EMBO journal, Vol. 19, Issue 23, pp. 6569-81, 2000 (PubMed).

Ajuh, Sleeman, Chusainow et al.: "A direct interaction between the carboxyl-terminal region of CDC5L and the WD40 domain of PLRG1 is essential for pre-mRNA splicing." in: The Journal of biological chemistry, Vol. 276, Issue 45, pp. 42370-81, 2001 (PubMed).

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