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Western blot analysis of PCNA was performed by loading 20 ug of HeLa (lane1), Raji (lane2), MCF-7 (lane3), Jurkat (lane4), HEK-293 (lane5), NIH/3T3 (lane6), MDA-MB-231 (lane7) cell lysate using Novex® NuPAGE® 4-12 % Bis-Tris gel (NP0322BOX), XCell SureLock™ Electrophoresis System (EI0002), Novex® Sharp Pre-Stained Protein Standard (LC5800), and iBlot® Dry Blotting System (IB21001). Proteins were transferred to a nitrocellulose membrane and blocked with 5 % skim milk for 1 hour at room temperature. PCNA was detected at ~ 34 kDa using PCNA, Biotin conjugate Mouse Monoclonal Antibody (133940) at 1:1000 dilution in 5 % skim milk at 4°C overnight on a rocking platform. Streptavidin - HRP (SA1007) at 1:2000 dilution was used and chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (WP20005).
|Tested species reactivity||Human, Insect, Mouse, Primate, Yeast|
|Published species reactivity||Sheep, Mouse|
|Host / Isotype||Mouse / IgG2a|
|Immunogen||Rat recombinant PCNA polypeptides.|
|Storage buffer||PBS, pH 7.4, with 50% glycerol, 1% BSA|
|Contains||0.1% sodium azide|
|Storage Conditions||4° C|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||3-5 µg/million cells|
|Immunohistochemistry (Paraffin) (IHC (P))||1:10-1:100|
|Western Blot (WB)||1:1000|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
PCNA is a marker for cells in early G1 phase and S phase of the cell cycle. It is found in the nucleus and is a cofactor of DNA polymerase delta. It acts as a homotrimer and helps increase the processivity of leading strand synthesis during DNA replication. In response to DNA damage, PCNA is ubiquitinated and is involved in the RAD6-dependent DNA repair pathway. Two transcript variants encoding the same protein have been found for PCNA. Pseudogenes of this gene have been described on chromosome 4 and on the X chromosome.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
The role of JNK in the development of hepatocellular carcinoma.
13-3940 was used in immunohistochemistry - paraffin section to elucidate JNK-regulated growth of hepatocellular carcinoma.
|Das M,Garlick DS,Greiner DL,Davis RJ||Genes & development (25:634)||2011|
|Mouse||Not Cited||Induction of KLF4 in basal keratinocytes blocks the proliferation-differentiation switch and initiates squamous epithelial dysplasia.||Foster KW,Liu Z,Nail CD,Li X,Fitzgerald TJ,Bailey SK,Frost AR,Louro ID,Townes TM,Paterson AJ,Kudlow JE,Lobo-Ruppert SM,Ruppert JM||Oncogene (24:1491)||2005|
|Sheep||Not Cited||Injury, inflammation, and remodeling in fetal sheep lung after intra-amniotic endotoxin.||Kramer BW,Kramer S,Ikegami M,Jobe AH||American journal of physiology. Lung cellular and molecular physiology (283:L452)||2002|
Anticancer activity of docetaxel in murine salivary gland carcinoma.
13-3940 was used in immunohistochemistry (paraffin) to assess the effects of docetaxel on salivary gland carcinoma.
|Piechocki MP,Lonardo F,Ensley JF,Nguyen T,Kim H,Yoo GH||Clinical cancer research : an official journal of the American Association for Cancer Research (8:870)||2002|
cyclin; DNA polymerase delta auxiliary protein; proliferating cell nuclear antigen