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, Non-human primate, Yeast|
|Published species reactivity||Sheep, Mouse, Human, Not Applicable|
|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.
|Immunohistochemistry (Paraffin) (IHC (P))||See 3 publications below|
|Immunohistochemistry (IHC)||See 1 publications below|
|Immunohistochemistry (Paraffin, Frozen) (IHC (P, F))||See 2 publications below|
|Miscellaneous PubMed (MISC)||See 1 publications below|
|Immunocytochemistry (ICC)||See 1 publications below|
|Western Blot (WB)||See 1 publications below|
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.
Oncogenic transformation of diverse gastrointestinal tissues in primary organoid culture.
13-3940 was used in immunohistochemistry - paraffin section to study the oncogenic transformation in primary organoid culture of gastrointestinal tissues
|Li X,Nadauld L,Ootani A,Corney DC,Pai RK,Gevaert O,Cantrell MA,Rack PG,Neal JT,Chan CW,Yeung T,Gong X,Yuan J,Wilhelmy J,Robine S,Attardi LD,Plevritis SK,Hung KE,Chen CZ,Ji HP,Kuo CJ||Nature medicine (20:769)||2014|
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 and development (25:634)||2011|
The CD4+ T cell-mediated IFN-gamma response to Helicobacter infection is essential for clearance and determines gastric cancer risk.
13-3940 was used in immunohistochemistry - paraffin section to describe a dual role for IFN-gamma in Helicobacter pathogenesis
|Sayi A,Kohler E,Hitzler I,Arnold I,Schwendener R,Rehrauer H,Müller A||Journal of immunology (Baltimore, Md. : 1950) (182:7085)||2009|
|Not Applicable||Not Cited||
Partial SP-B deficiency perturbs lung function and causes air space abnormalities.
13-3940 was used in immunohistochemistry to use transgenic mice to investigate how surfactant protein B deficiency perturbs pulmonary function
|Nesslein LL,Melton KR,Ikegami M,Na CL,Wert SE,Rice WR,Whitsett JA,Weaver TE||American journal of physiology. Lung cellular and molecular physiology (288:L1154)||2005|
|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|
||The 104-123 amino acid sequence of the beta-domain of von Hippel-Lindau gene product is sufficient to inhibit renal tumor growth and invasion.||Datta K,Sundberg C,Karumanchi SA,Mukhopadhyay D||Cancer research (61:1768)||2001|
|Human||Not Cited||The 104-123 amino acid sequence of the beta-domain of von Hippel-Lindau gene product is sufficient to inhibit renal tumor growth and invasion.||Datta K,Sundberg C,Karumanchi SA,Mukhopadhyay D||Cancer research (61:1768)||2001|