- 51-0700 - Invitrogen Antibodies YWHAZ antibody

Submitted references Parkin Coordinates Platelet Stress Response in Diabetes Mellitus: A Big Role in a Small Cell.

Lee SH, Du J, Hwa J, Kim WH
International journal of molecular sciences 2020 Aug 15;21(16)

Junctional tumor suppressors interact with 14-3-3 proteins to control planar spindle alignment.

Nakajima YI, Lee ZT, McKinney SA, Swanson SK, Florens L, Gibson MC
The Journal of cell biology 2019 Jun 3;218(6):1824-1838

Protein Phosphatase (PP2C9) Induces Protein Expression Differentially to Mediate Nitrogen Utilization Efficiency in Rice under Nitrogen-Deficient Condition.

Waqas M, Feng S, Amjad H, Letuma P, Zhan W, Li Z, Fang C, Arafat Y, Khan MU, Tayyab M, Lin W
International journal of molecular sciences 2018 Sep 19;19(9)

14-3-3 proteins regulate Tctp-Rheb interaction for organ growth in Drosophila.

Le TP, Vuong LT, Kim AR, Hsu YC, Choi KW
Nature communications 2016 May 6;7:11501

14-3-3 Epsilon antagonizes FoxO to control growth, apoptosis and longevity in Drosophila.

Nielsen MD, Luo X, Biteau B, Syverson K, Jasper H
Aging cell 2008 Oct;7(5):688-99

14-3-3zeta is indispensable for aggregate formation of polyglutamine-expanded huntingtin protein.

Omi K, Hachiya NS, Tanaka M, Tokunaga K, Kaneko K
Neuroscience letters 2008 Jan 24;431(1):45-50

Proteomic analysis of bovine sperm YWHA binding partners identify proteins involved in signaling and metabolism.

Puri P, Myers K, Kline D, Vijayaraghavan S
Biology of reproduction 2008 Dec;79(6):1183-91

Phosphorylation-dependent interaction of tyrosine 3-monooxygenase/tryptophan 5-monooxygenase activation protein (YWHA) with PADI6 following oocyte maturation in mice.

Snow AJ, Puri P, Acker-Palmer A, Bouwmeester T, Vijayaraghavan S, Kline D
Biology of reproduction 2008 Aug;79(2):337-47

C-terminal recognition by 14-3-3 proteins for surface expression of membrane receptors.

Coblitz B, Shikano S, Wu M, Gabelli SB, Cockrell LM, Spieker M, Hanyu Y, Fu H, Amzel LM, Li M
The Journal of biological chemistry 2005 Oct 28;280(43):36263-72

No comments: Submit comment

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of HeLa cells using Rb anti-14-3-3 (Product # 51-0700). Lane 1: 1 µg/mL, Lane 2: 0.5, g/mL, Lane 3: 0.1 µg/mL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis was performed on whole cell extracts (30 µg) of A-431 (Lane 1), HCT 116 (Lane 2), HEK-293 (Lane 3), HeLa (Lane 4), SH-SY5Y (Lane 5), NIH/3T3 (Lane 6) and PC-12 (Lane 7). The blots were probed with Anti- 14-3-3 theta Rabbit Polyclonal Antibody (Product # 51-0700, 2 µg/mL) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conj µgate (Product # A27036, 0.4 µg/mL, 1:2500 dilution). A ~ 28 kDa band corresponding to 14-3-3 theta was observed across cell lines tested. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris gel (Product # NP0321BOX), 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 iBlot® 2 Dry Blotting System (Product # IB21001). 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: Immunofluorescence analysis of 14-3-3 theta was performed using 70% confluent log phase MCF-7 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 with 14-3-3 theta Rabbit Polyclonal Antibody (Product # 51-0700) at 2 µg/mL in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conj µgate (Product # A27034) 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 represents the merged image showing cytoplasmic localization. Panel e shows the no primary antibody control. The images were captured at 60X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Flow Cytometry analysis of 14-3-3 theta was done on MCF7 cells (untreated, red histogram) and MCF7 cells treated with 100uM Etoposide for 6 hours (blue histogram). 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 theta Rabbit Polyclonal Antibody (Product # 51-0700) or with rabbit isotype control (pink histogram) at 3-5 µg/million cells in 2.5% BSA. After incubation at room temperature for 2 hours, the cells were labeled with Alexa Fluor® 488 Goat Anti-Rabbit Secondary Antibody (Product # A11008) 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.

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: NULL

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: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 3 Effects of Tctp mutation on Rheb and 14-3-3s. ( a - d ) Clones of Tctp h59 mutant cells in eye disc. ( e - h ) Clones of Tctp h59 mutant cells in wing disc. These clones are marked by the absence of Arm-lacZ. In both organs, Tctp mutant clones show reduction of Rheb ( c , g ) but not 14-3-3s ( d , h ). Red rectangles in a and e indicate the enlarged regions shown in b - d and f - h respectively. Scale bar, 100 mum ( a - h ).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 5 14-3-3s are required for the interaction between Tctp and Rheb and organ development. ( a ) Knockdown of 14-3-3s in S2 cells shows depletion of 14-3-3s levels after 6 days of RNAi treatment. ei, zetai and ei+zetai indicate RNAi for 14-3-3e, 14-3-3zeta and both isoforms, respectively. Levels of Tctp and Rheb were not noticeably affected by 14-3-3 depletion. The treatment performed to detect Tctp, Rheb, and actin level was double knockdown of both isoforms of 14-3-3. ( b ) 14-3-3 depletion affects the interaction between V5-Tctp and V5myc-Rheb after 6 days of RNAi treatment. Anti-Myc antibody was used for IP. Tctp was detected by anti-Tctp antibody. Depletion of both isoforms abolished the Tctp-Rheb interaction. ( c - j ) Double knockdown of 14-3-3s causes loss of targeted tissues. ey-Gal4 control ( c ) 14-3-3e RNAi ( d ) and 14-3-3zeta RNAi ( e ) show normal pattern of eye-head in pupae. Knockdown of both 14-3-3 isoforms causes pupal lethality with loss of eye and head ( f ). ( g - j ) Pattern of Elav staining in larval eye discs from indicated genotypes. Knockdown of 14-3-3e or 14-3-3zeta has no obvious effect on eye disc development ( h , i ). Knockdown of both isoforms results in small disc (the area marked with dotted line) with no retinal differentiation ( j ). ( k - n ) Double knockdown of 14-3-3s results in reduced proliferation. MS1096-Gal4 control ( k ) 14-3-3e RNAi ( l ) and 14-3-3zeta RNAi ( m ) show normal level of PH3 staining in wing disc. Knockdown of b

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 6 Effects of 14-3-3 RNAi on Tor targets and the suppression by CycE. ( a ) Effects of 14-3-3 RNAi on S6k and Thor. Either one of 14-3-3 isoforms or both 14-3-3s were knocked down in S2 cells. Single or double knockdown of 14-3-3s reduced the level of pS6k (Thr 398) but does not affect the level of S6k protein. Knockdown of both 14-3-3s, but not one isoform, causes slight reduction of Thor. In contrast, pThor (Thr 37/46) is strongly reduced by single or double knockdown of 14-3-3s. ( b - d ) Loss of 14-3-3s results in reduction of CycE level. ( b ) 14-3-3e mutant clones induced by ey-flp are marked by the absence of GFP. Mutant clones show strong reduction of CycE ( c ). ( e - g ) 14-3-3zeta mutant clones induced by hs-flp are marked by the absence of Arm-lacZ. CycE is reduced in mutant clones ( f ). ( h - m ) Effects of CycE in eye. Control with one copy of ey-Gal4 ( h ) or overexpression of CycE ( i ) shows normal eye. ( j ) Double knockdown of Tctp and 14-3-3 results in small and rough eye phenotype (Tctp RNAi/ CycE GFP ; 14-3-3 RNAi/+ were scored. ( o - r ) CycE suppresses the wing pheno

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 9 ( A )Western blotting of differentially expressed proteins in PP2C9TL and WT at 10 DAF stage. The top photo shows the SDS-PAGE separation of two protein samples used as loading control. The bottom photo shows gel transferred onto nitrocellulose membrane for western blotting to detect actin, anti-Flag- PP2C9 , and 14-3-3 protein for WT and PP2C9TL at the 10 DAF stage. ( B ) Co-immunoprecipitation (Co-IP) assays of PP2C9 associated protein analyzed by SDS-PAGE. Protein molecular weight marker, negative control without antibodies, WT protein, and the third is the PP2C9TL protein. 2.8. Identification of PP2C9 Associated Protein by Co-Immunoprecipitation (CO-IP) and Mass Spectrometry.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 3 Confirmation of interactions between Parkin and selected target proteins in human and murine platelets. ( A ) Immunoprecipitation of each specific antibody in pooled human DM platelets (4 HC, 5DM1, and 6DM2). We incubated 500 mug protein lysates incubated with specific antibodies overnight at 4 degC with 10% input as control. G1 represents interacting protein groups from LC-MS/MS results. G2 represents the non-interacting proteins group in LC-MS/MS results. G3 represents protein that interacted with Parkin and that were not found in the LC-MS/MS results. ( B ) Immunoprecipitation of each specific antibody in pooled murine platelets (3 WT and 5DM). We incubated 500 mug protein lysates incubated with specific antibodies overnight at 4 degC with 10% input as control.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 4. Drosophila 14-3-3 proteins interact with Dlg in the wing disc epithelium. (A) Coimmunoprecipitation of endogenous 14-3-3 proteins and Dlg from wild-type wing disc samples. Western blots were probed with anti-14-3-3 antibody (upper lanes) and anti-Dlg antibody (lower lanes) in input and immunoprecipitates pulled down by anti-Dlg antibody. IP, immunoprecipitation. (B-G) PLA indicates close proximity or interactions of two proteins. PLA between Scrib and Dlg in control ( nub-Gal4/ +; B) and dlg -knockdown ( nub-Gal4>dlg-RNAi ; C) wing discs. (D) Quantification of the number of PLA (Scrib/Dlg) spots for control ( n = 6) and dlg-RNAi ( n = 5) wing discs. PLA between 14-3-3 and Dlg in control (E) and dlg -knockdown (F) wing discs. (G) Quantification of the number of PLA (14-3-3/Dlg) spots for control ( n = 5) and dlg-RNAi ( n = 5) wing discs. Error bars are SD. **, P < 0.01 by Kolmogorov-Smirnov test. Note that PLA signals in MZ of the disc proper diminished in dlg -RNAi wing discs, but not in PE where nub-Gal4 is not expressed. PE, peripodial epithelium; MZ, mitotic zone. (H and I) Subcellular localization of 14-3-3 proteins detected by anti-14-3-3 antibody staining. 14-3-3 proteins localize to the apical cortex and nucleus during interphase (H) and cytosol, including spindle microtubules (alpha-tubulin), during mitosis (I). Yellow arrows indicate metaphase cells. Scale bars: 5 um.

51-0700

Antibody data