- MA5-11663 - Invitrogen Antibodies SFN antibody

Submitted references Activation of the Ca(2+) sensing receptor and the PKC/WNK4 downstream signaling cascade induces incorporation of ZO-2 to tight junctions and its separation from 14-3-3.

Amaya E, Alarcón L, Martín-Tapia D, Cuellar-Pérez F, Cano-Cortina M, Ortega-Olvera JM, Cisneros B, Rodriguez AJ, Gamba G, González-Mariscal L
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Hypoxia induces an undifferentiated phenotype of oral keratinocytes in vitro.

Kato H, Izumi K, Uenoyama A, Shiomi A, Kuo S, Feinberg SE
Cells, tissues, organs 2014;199(5-6):393-404

Identification of novel target proteins in sebaceous gland carcinoma.

Erovic BM, Al Habeeb A, Harris L, Goldstein DP, Kim D, Ghazarian D, Irish JC
Head & neck 2013 May;35(5):642-8

14-3-3 sigma is a useful immunohistochemical marker for diagnosing ovarian granulosa cell tumors and steroid cell tumors.

Chen L, Yang B
International journal of gynecological pathology : official journal of the International Society of Gynecological Pathologists 2013 Mar;32(2):156-62

Prognostic and predictive markers in medullary thyroid carcinoma.

Erovic BM, Kim D, Cassol C, Goldstein DP, Irish JC, Asa SL, Mete O
Endocrine pathology 2012 Dec;23(4):232-42

Biomarkers of parathyroid carcinoma.

Erovic BM, Harris L, Jamali M, Goldstein DP, Irish JC, Asa SL, Mete O
Endocrine pathology 2012 Dec;23(4):221-31

Novel snail1 target proteins in human colon cancer identified by proteomic analysis.

Larriba MJ, Casado-Vela J, Pendás-Franco N, Peña R, García de Herreros A, Berciano MT, Lafarga M, Casal JI, Muñoz A
PloS one 2010 Apr 20;5(4):e10221

Proteomic profiling of differential display analysis for human oral squamous cell carcinoma: 14-3-3 σ Protein is upregulated in human oral squamous cell carcinoma and dependent on the differentiation level.

Hayashi E, Kuramitsu Y, Fujimoto M, Zhang X, Tanaka T, Uchida K, Fukuda T, Furumoto H, Ueyama Y, Nakamura K
Proteomics. Clinical applications 2009 Nov;3(11):1338-47

Loss of 14-3-3 sigma protein expression and presence of human papillomavirus type 16 E6 in oral squamous cell carcinoma.

Bhawal UK, Sugiyama M, Nomura Y, Kuniyasu H, Tsukinoki K
Archives of otolaryngology--head & neck surgery 2008 Oct;134(10):1055-9

The clinicopathological and prognostic impact of 14-3-3 sigma expression on vulvar squamous cell carcinomas.

Wang Z, Tropè CG, Suo Z, Trøen G, Yang G, Nesland JM, Holm R
BMC cancer 2008 Oct 24;8:308

Immunostaining as an adjunct to cytology for diagnosis of pancreatic adenocarcinoma.

Agarwal B, Ludwig OJ, Collins BT, Cortese C
Clinical gastroenterology and hepatology : the official clinical practice journal of the American Gastroenterological Association 2008 Dec;6(12):1425-31

p63 Is essential for the proliferative potential of stem cells in stratified epithelia.

Senoo M, Pinto F, Crum CP, McKeon F
Cell 2007 May 4;129(3):523-36

Growth inhibitory effect of the human NIT2 gene and its allelic imbalance in cancers.

Lin CH, Chung MY, Chen WB, Chien CH
The FEBS journal 2007 Jun;274(11):2946-56

Promoter methylation of PARG1, a novel candidate tumor suppressor gene in mantle-cell lymphomas.

Ripperger T, von Neuhoff N, Kamphues K, Emura M, Lehmann U, Tauscher M, Schraders M, Groenen P, Skawran B, Rudolph C, Callet-Bauchu E, van Krieken JH, Schlegelberger B, Steinemann D
Haematologica 2007 Apr;92(4):460-8

Promoter methylation of PARG1, a novel candidate tumor suppressor gene in mantle-cell lymphomas.

Ripperger T, von Neuhoff N, Kamphues K, Emura M, Lehmann U, Tauscher M, Schraders M, Groenen P, Skawran B, Rudolph C, Callet-Bauchu E, van Krieken JH, Schlegelberger B, Steinemann D
Haematologica 2007 Apr;92(4):460-8

The p53 pathway and outcome among patients with T1G3 bladder tumors.

López-Knowles E, Hernández S, Kogevinas M, Lloreta J, Amorós A, Tardón A, Carrato A, Kishore S, Serra C, Malats N, Real FX, EPICURO Study Investigators
Clinical cancer research : an official journal of the American Association for Cancer Research 2006 Oct 15;12(20 Pt 1):6029-36

Identification of 14-3-3sigma as a contributor to drug resistance in human breast cancer cells using functional proteomic analysis.

Liu Y, Liu H, Han B, Zhang JT
Cancer research 2006 Mar 15;66(6):3248-55

Expression and prognostic significance of 14-3-3sigma and ERM family protein expression in periampullary neoplasms.

Hustinx SR, Fukushima N, Zahurak ML, Riall TS, Maitra A, Brosens L, Cameron JL, Yeo CJ, Offerhaus GJ, Hruban RH, Goggins M
Cancer biology & therapy 2005 May;4(5):596-601

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot of 14-3-3 sigma using 14-3-3 sigma Monoclonal Antibody (Product # MA5-11663) on HT29 Cells.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot of 14-3-3 sigma using 14-3-3 sigma Monoclonal Antibody (Product # MA5-11663) on HT29 Cells.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis was performed on whole cell extracts (30 µg lysate) of A549 (lane 1), MCF-7 (lane 2), A431 (lane 3), HT-29 (Lane 4) and HeLa (lane 5). The blots were probed with Anti-14-3-3 Sigma Mouse Monoclonal Antibody (Product # MA5-11663, 0.5-1 µg/mL) and detected by chemiluminescence using Goat anti-Mouse IgG (H+L) Secondary Antibody, HRP conjugate (Product # 62-6520, 1:4000 dilution). A 28 kDa band corresponding to 14-3-3 Sigma was observed across cell line tested. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 10 % Bis-Tris gel (Product # NP0301BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with PierceTM Power Blotter System (Product # 22834) The membrane was probed with the relevant primary and secondary Antibody following blocking with 5 % skimmed milk. Chemiluminescent detection was performed using Pierce™ ECL Western Blotting Substrate (Product # 32106).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of 14-3-3 sigma (green) showing staining in the cytoplasm of A431 cells (right) compared to a negative control without primary antibody (left). Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were probed with a 14-3-3 sigma monoclonal antibody (Product # MA5-11663) in 3% BSA-PBS at a dilution of 1:100 and incubated overnight at 4 ºC in a humidified chamber. Cells were washed with PBST and incubated with a DyLight-conjugated secondary antibody in PBS at room temperature in the dark. F-actin (red) was stained with a fluorescent red phalloidin and nuclei (blue) were stained with Hoechst or DAPI. Images were taken at a magnification of 60x.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of 14-3-3 sigma (green) showing staining in the cytoplasm of Hela cells (right) compared to a negative control without primary antibody (left). Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were probed with a 14-3-3 sigma monoclonal antibody (Product # MA5-11663) in 3% BSA-PBS at a dilution of 1:100 and incubated overnight at 4 ºC in a humidified chamber. Cells were washed with PBST and incubated with a DyLight-conjugated secondary antibody in PBS at room temperature in the dark. F-actin (red) was stained with a fluorescent red phalloidin and nuclei (blue) were stained with Hoechst or DAPI. Images were taken at a magnification of 60x.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis of 14-3-3 sigma was done on 70% confluent log phase HeLa cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled 14-3-3 sigma (1433S01) Mouse Monoclonal Antibody (Product # MA5-11663) at 2 µg/mL in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A28175) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Alexa Fluor® 555 Rhodamine Phalloidin (Product # R415, 1:300). Panel d is a merged image showing cytoplasmic localization. Panel e is a no primary antibody control. The images were captured at 60X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Formalin-fixed, paraffin-embedded human skin stained with 14.3.3 sigma antibody using peroxidase-conjugate and AEC chromogen. Note cytoplasmic staining of epidermis.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Flow cytometry analysis of 14-3-3 sigma was done on A-431 cells. Cells were fixed with 70% ethanol for 10 minutes, permeabilized with 0.25% Triton™ X-100 for 20 minutes, and blocked with 5% BSA for 30 minutes at room temperature. Cells were labeled with 14-3-3 sigma Mouse Monoclonal Antibody (MA511663, red histogram) or with mouse isotype control (yellow histogram) at 3-5 ug/million cells in 2.5% BSA. After incubation at room temperature for 2 hours, the cells were labeled with Alexa Fluor® 488 Rabbit Anti-Mouse Secondary Antibody (A11059) at a dilution of 1:400 for 30 minutes at room temperature. The representative 10,000 cells were acquired and analyzed for each sample using an Attune® Acoustic Focusing Cytometer. The purple histogram represents unstained control cells and the green histogram represents no-primary-antibody control.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: FIGURE 9: The cellular content of ZO-2 diminishes after the CS due to lysosome-mediated degradation. (A) The amount of ZO-2, 14-3-3sigma, and 14-3-3zeta diminishes with the CS. Cell lysates were obtained from monolayers cultured in the LC condition for 20 h or 2, 4, and 6 h after a CS. Top panels, representative Western blots; bottom panels, densitometric analysis. Statistical analysis done with One Way ANOVA followed by Dunnett's multiple comparison test. ns, not significant; * p < 0.05; ** p < 0.01; *** p < 0.001. All the results in this figure correspond to at least three independent experiments. (B) The decay of ZO-2 triggered by the CS can be reversed with cloroquine but not with MG132. Cell lysates were obtained from monolayers cultured in the LC condition for 20 h and incubated or not for 5 additional h with 30 muM cycloheximide (CHX) or transferred to NC media (CS) for 5 h in the presence or absence of 30 muM cycloheximide (CHX) with or without 30 muM MG132 or 50 muM cloroquine. Top panel, representative Western blot; bottom panel, densitometric analysis. Statistical analysis done with one way ANOVA followed by Dunnett's multiple comparison test; ns, not significant; *** p < 0.001; ** p < 0.01; * p < 0.05. (C) The 14-3-3sigma and 14-3-3zeta are degraded in the proteosome and not in the lysosome during the CS. Cell lysates were obtained from monolayers cultured in the LC condition for 20 h or transferred to NC media (CS) for 6 h in the presence or absence of 30 muM MG1

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: FIGURE 11: The U2 segment of ZO-2 favors the interaction of ZO-2 with 14-3-3. MDCK monolayers cultured in LC medium were transfected with WT hZO-2 (Flag-hZO-2) or a hZO-2 lacking the U2 region (Flag-hZO-2 DeltaU2) and treated with 30 muM MG132 for 4 h. (A) A PLA assay was done employing a rabbit antibody against Flag and mouse antibodies against 14-3-3sigma or 14-3-3zeta. Transfected cells were identified by their green staining after treatment with an antibody against rabbit coupled to Alexa Fluor 488. Background, untransfected cells. Bars, 15 mum. Top panel, representative images; bottom panel, quantitative analysis was done using BlobFinder. Statistical analysis done with one-way ANOVA followed by Dunnett's multiple comparison test; *** p < 0.001; **** p < 0.0001. Results were obtained analyzing 20 transfected cells per condition derived from two independent experiments. (B) Immunoprecipitation assay was done with an antibody against Flag and blotted with antibodies against Flag, 14-3-3sigma, and 14-3-3zeta. PIS, preimmune serum. Left panels, representative images; right panels, quantitative analysis of three independent experiments. Statistical analysis done with student's t test; * p < 0.05; ** p < 0.01.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: FIGURE 13: cZO-2 mutants S261A and T248A displays reduced binding to 14-3-3sigma and 14-3-3zeta. (A) The interaction of endogenous 14-3-3sigma and transfected HA-cZO-2 diminishes with cZO-2 mutants S261A and T248A. Cells were transfected with WT HA-cZO-2 or the HA-cZO-2 mutants T248A and S261A. PLA was done with a rabbit antibody against HA and a mouse antibody against 14-3-3sigma in the presence of MG132. In this experiment, the cells transfected with HA-cZO-2 or the HA-cZO-2 mutants T248A and S261A were identified with a rabbit antibody against HA, followed by a secondary anti rabbit antibody coupled to Alexa Fluor 488. Background corresponds to cells not transfected with HA-cZO-2. Bars, 15 mum. Left panel, representative images; right panel, quantitative analysis done using BlobFinder. Statistical analysis was done with Kruskal-Wallis test followed by Dunn's multiple comparison test; * p < 0.05; **** p < 0.0001. Results were obtained with 30 transfected cells per condition derived from two independent experiments. (B) The interaction of endogenous 14-3-3zeta with transfected HA-cZO-2 diminished in mutants cZO-2 S261A and T248A. Cells were transfected with HA-cZO-2 or the HA-cZO-2 mutants T248A and S261A. PLA were done in the presence of MG132 with a rabbit antibody against HA and a mouse antibody against 14-3-3zeta. In this experiment, the transfected cells were identified with a rabbit antibody against HA, followed by a secondary anti rabbit antibody coupled to Alexa Fluo

MA5-11663

Antibody data