13-8500

antibody from Invitrogen Antibodies

Targeting: JUP CTNNG, DP3, DPIII, PDGB, PKGB

Provider product page for 13-8500

Western blot

Immunocytochemistry

Immunoprecipitation

Immunohistochemistry

Flow cytometry

Other assay

Antibody data

Antibody Data
Antigen structure
References [26]
Comments [0]
Validations
Immunocytochemistry [1]
Flow cytometry [1]
Other assay [17]

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Product number: 13-8500 - Provider product page
Provider: Invitrogen Antibodies
Product name: gamma Catenin Monoclonal Antibody (PG-11E4)
Antibody type: Monoclonal
Antigen: Other
Reactivity: Human, Mouse
Host: Mouse
Isotype: IgG
Antibody clone number: PG-11E4
Vial size: 100 µg
Concentration: 0.5 mg/mL
Storage: -20°C

JUP protein structure - 13-8500 shown in red.

Supportive validation

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

Supportive validation

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

Supportive validation

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

Supportive validation

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

Supportive validation

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

Supportive validation

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

Supportive validation

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

Supportive validation

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

Supportive validation

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

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 1 Plakoglobin binds to SOX4 in LNCaP cells . A . Whole cell lysates prepared from LNCaP cells transfected with pREP4-BLRPwt-HASOX4-IRES-BirA-XL9 or vector only were purified using an streptavidin-magnetic beads, and affinity-purified Vector- and SOX4-complexes were visualized by silver staining. SOX4 protein is indicated by an arrow. The molecular weight markers are as shown on the left. B . Amino acid sequence of plakoglobin with trypsinized peptides from LC-MS/MS analysis indicated in bold and underlined. C , pREP4-BLRPwt-HASOX4-IRES-BirA-XL9 and vector control transfected LNCaP cells. SOX4-associated plakoglobin were analyzed by Western blot with antibodies indicated. Equivalent amounts of the Vector-purified fractions were used to confirm specificity. 5% of transfected LNCaP whole cell lysate was used as Input. D , endogenous plakoglobin IPs were analyzed for transfected SOX4 by Western blot with anti-HA 16B12 mAb. E , endogenous plakoglobin IPs were analyzed for endogenous SOX4 in untransfected LNCaP cells. F, G, endogenous plakoglobin IPs were analyzed for endogenous SOX4 in primary human keratinocytes and breast cancer cell line, MDA-MB-231, respectively. H, I, HASOX4 IPs were analyzed for endogenous plakoglobin in stably-expressed SOX4 PC3M and LNCaP cell lines. F-I, whole cell lysates were harvested and treated with DNase I for 1 hr at room temperature prior to immunoprecipitation.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 2 Wnt signaling induces nuclear colocalization of SOX4 and plakoglobin . Subcellular localization of plakoglobin and SOX4 were examined by confocal microscopy. PC3M HASOX4 stable cell line was treated with 100 ng/ml human recombinant WNT3A or 20 muM leptomycin b (LMB), or both WNT3A+LMB for 24 hr. The fields shown were visualized independently by confocal microscopy at the appropriate wavelength for plakoglobin (488) and SOX4 (543), and Topro (633) respectively, and then the three images were overlaid (Merge). Strong nuclear localization of plakoglobin was observed in the WNT3A+LMB treated cells that expressed HASOX4. Representative fields from these independent repeated experiments are shown. Plakoglobin localizes to the nucleus following WNT3A treatment, and this effect is strongly enhanced by LMB co-treatment, suggesting shuttling of plakoglobin into and out of the nucleus following WNT3A stimulation. Note that plakoglobin nuclear localization is much stronger in cells expressing HASOX4, suggesting SOX4 may facilitate plakoglobin nuclear import.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 3 Interaction between SOX4 and plakoglobin is affected by Wnt3A signaling . A . LNCaP HASOX4 stable cell lines were treated with vehicle, Wnt3A, LMB, or Wnt3A + LMB and anti-HA immunoprecipitations were immunoblotted for the presence of plakoglobin. B , Quantitative analysis of co-IP and Western blots from panel A using the Odyssey (r) Infrared Imaging System (Li-Cor Bioscience). Fold changes were calculated as the ratio of immunoprecipitated plakoglobin to SOX4 relative to the vehicle control panel (untreat). (n = 3, independent biological replicates performed on separate days; error bars represent SEM; *, p < 0.05; **, p < 0.01 for a one-sided paired t-test) C . (Upper) LNCaP HASOX4 stable cell lines were treated with vehicle, WNT3A, LMB, or WNT3A + LMB and anti-HA immunoprecipitations were performed using separate cytosolic and nuclear fractions. (Lower) Anti-AKT and anti-Lamin A antibodies were used as controls to examine markers of cytosolic and nuclear fractions, respectively, to confirm the purity of each fraction. D . Quantitative analysis of co-IP and Western blots from panel C.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 4 SOX4 transcriptional activity is modulated by Wnt-induced interaction with plakoglobin . A . ChIP assay of HA-SOX4 bound to the predicted SOX4-binding sites on DICER1 , SOX4 , DHX9 , and AXIN2 promoters. GAPDH is shown as a negative control. B . Quantitative results of ChIP by realtime-PCR. SOX4 promoter occupancy is increased with Wnt treatment, but is strongly reduced by combined WNT3A+LMB treatment. (n = 4; error bars represent SEM; *, p < 0.05 for a one-sided paired t-test) C . SOX4 targets and Wnt downstream genes are inhibited after Wnt induction and LMB treatment. Realtime-PCR expression analysis of SOX4 direct targets and Wnt signaling downstream genes after 24 hrs of WNT3A and LMB treatment. D . siRNA directed against plakoglobin downregulates protein expression in LNCaP HA-SOX4 cell lines. Cells were harvested 48-hr post-transfection and Western blot were probed with anti-plakoglobin and anti-beta-actin antibodies in the presence or absence of WNT3A treatment. E . Quantitative ChIP-qPCR assay of HA-SOX4 following plakoglobin knockdown. HA-SOX4 promoter occupancy is strongly reduced by combined WNT3A and LMB treatment but is partially restored when plakoglobin is knocked down by siRNA treatment. Control scrambled siRNA had no effect (n = 3; error bars represent SEM; *, p < 0.05 for a one-sided paired t-test).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Fig. 3 Activation of autophagy during syncytialization. a Primary trophoblastic cells were labeled with acridine orange 24, 48, 72 and 96 h after seeding and observed by fluorescence microscopy. Scale bar represents 100 mum. b Acridine orange quantification of the intensity and area of red signal, normalized to the total number of nuclei. n = 4. Data represented as mean +- SEM. ** P

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Fig. 4 Consequences of autophagy modulation with autophagy inhibitors or activators on trophoblastic cell fusion and differentiation. vCTB were purified from human term placenta and seeded for 24 h prior treatment with 10 nM Bafilomycin A1 (BAF), 2 uM Chloroquine (CQ), 10 nM Trichostatin A (TRI), 200 uM Valproic acid (VA), or Ethanol (Control Ethanol, Cnt Eth) for 48 h. a , b Western blotting was performed on the cells. LC3b-II and GAPDH levels were quantified using the ImageJ software, and data are expressed as the fold change relative to the control. n = 4. Data represented as mean +- SEM. * P

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Fig. 6 Effect of UPR modulation on autophagy activation in trophoblastic cells. vCTB were purified from human term placenta and seeded for 24 h prior treatment with 100 nM GSK2656157 (GSK), 100 uM STF-083010 (STF), 200 uM 4-(2-aminoethyl)benzenesulfonyl fluoride hydrochloride (AEBSF), or DMSO (Control DMSO, Cnt DMSO) for 48 h. a Cells were labeled with acridine orange 48 h post-treatment and observed by fluorescence microscopy. Scale bar represents 100 mum. b Acridine orange quantification of the intensity and area of red signal, normalized to the total number of nuclei. n = 4. Data represented as mean +- SEM. * P

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 2 Solubility assays . Solubility patterns for (A) desmosomal and (B) nuclear envelope proteins were assessed by immunoblots for soluble and insoluble protein fractions extracted from COS-7 cells transfected with wildtype (WT) TMEM43 , mutant (Mut) TMEM43 , or a 1:2 ratio of wildtype to mutant (WT + Mut) TMEM43 . Sol, soluble fraction containing cytoplasmic (Cyt) proteins; Insol, insoluble fraction containing membrane bound proteins. No differences in solubility patterns were found, suggesting that the presence of mutant TMEM43 did not alter the stability of TMEM43 or of other desmosomal or nuclear envelope proteins.