14-3-3 eta Antibody

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14-3-3 eta Antibody gallery image 1

Western blots of 14-3-3 isoforms in HEK293 cell extracts using Bio-Rad antibodies.
Lane 1 = anti-gamma 14-3-3 (AHP1047)
Lane 2 = anti-zeta 14-3-3 (AHP1052)
Lane 3 = anti-C-epsilon-14-3-3 (AHP1045)
Lane 4 = anti-N-epsilon-14-3-3 (AHP1048)
Lane 5 = anti-eta 14-3-3. (AHP1046)
The 14-3-3 isoforms run at 30 kDa except epsilon 14-3-3 which migrates at 33 kDa

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Immunoblots identifying 14-3-3 isoforms in extracts from mouse oocytes, eggs and ovaries. Proteins from cell lysates were separated by electrophoresis under reducing conditions, transferred to membranes and probed with antibodies directed against the 14-3-3 isoforms indicated. Each isoform shown was detected in lysates of 200 oocytes or 200 eggs. Protein extracts of ovaries and brain from adult mice are included for comparison. The 14-3-3 protein is approximately 30 kDa.

From: De S, Marcinkiewicz JL, Vijayaraghavan S, Kline D. Expression of 14-3-3 protein isoforms in mouse oocytes, eggs and ovarian follicular development. BMC Res Notes. 2012 Jan 23;5:57.

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Representative immunofluorescence images of 14-3-3 isoforms in oocytes and eggs isolated from adult mice. (A, B) 14-3-3β. (C, D) 14-3-3γ. (E, F) 14-3-3ε. (G, H) 14-3-3ζ. (I, J) 14-3-3 τ. (K, L) 14-3-3σ. (M, N) 14-3-3η. Confocal sections with regions of red fluorescence indicating the corresponding isoforms studied (see Methods). The inset in N shows the same egg labeled blue with Hoechst DNA stain (non-confocal image) and confirms that the darker areas in this region of the larger image are condensed metaphase II chromosomes. Control cells were included for each isoform experiment and were imaged using the same confocal settings. Representative control oocytes and eggs are shown in bright-field (O) and fluorescence (P). 14-3-3? accumulates, in part, in the meiotic spindle in eggs as shown by simultaneous labeling with 14-3-3? (Q) and tubulin (R) antibodies. These sequential scans are merged (14-3-3? + tubulin) in (S). The scale bars represent 10 µm for all images.

From: De S, Marcinkiewicz JL, Vijayaraghavan S, Kline D. Expression of 14-3-3 protein isoforms in mouse oocytes, eggs and ovarian follicular development. BMC Res Notes. 2012 Jan 23;5:57.

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Representative immunohistochemistry images of 14-3-3 η in the different stages of follicular development in ovarian sections. (A) Primordial follicle. (B) Primary follicle. (C) Secondary follicle. (D) Early antral follicle. (E) Graafian (advanced antral) follicle. (F) Corpus luteum. White arrows indicate the primordial or primary follicles in (A and B). Note the weaker staining in mural granulosa cells in secondary follicles (C, green arrows) and the more intense staining in cells lining the antral cavity (E, red arrows). The scale bars represent 10 µm (A-C) or 100 µm (D-F).

From: De S, Marcinkiewicz JL, Vijayaraghavan S, Kline D. Expression of 14-3-3 protein isoforms in mouse oocytes, eggs and ovarian follicular development. BMC Res Notes. 2012 Jan 23;5:57.

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Representative immunohistochemistry images of 14-3-3 protein isoforms in atretic follicles of adult mouse ovaries. (A) 14-3-3 β. (B) 14-3-3 γ. (C) 14-3-3 ε. (D) 14-3-3 ζ. (E) 14-3-3η. (F) 14-3-3 τ. (G) 14-3-3 σ. All scale bars indicate 100 µm.

From: De S, Marcinkiewicz JL, Vijayaraghavan S, Kline D. Expression of 14-3-3 protein isoforms in mouse oocytes, eggs and ovarian follicular development. BMC Res Notes. 2012 Jan 23;5:57.

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Representative immunocytochemistry images of 14-3-3 protein isoforms along and in the zonae pellucidae of cumulus-free oocytes isolated from ovaries of adult mice. The zona-intact cells were fixed in paraformaldehyde but not treated with detergent (see Methods). Paired images of an oocyte (left image is brightfield and right is immunofluorescence) indicate staining along the zona and/or the cell membrane for all isoforms except 14-3-3ζ. Note: the intent of this part of the study was not to examine the intracellular distribution of the isoforms (see Figure 3 for those experiments) since the cells were not permeabilized to permit complete antibody penetration; however, cells may be partially permeabilized by fixation accounting for the detection of intracellular proteins in some cells. (A,B) 14-3-3 ß. (C,D) 14-3-3 ?. (E,F) 14-3-3 e. (G,H) 14-3-3 ?. (I,J) 14-3-3 ?. (K,L) 14-3-3 t. (M,N) 14-3-3s. (O,P) Control image without primary antibody addition.

From: De S, Marcinkiewicz JL, Vijayaraghavan S, Kline D. Expression of 14-3-3 protein isoforms in mouse oocytes, eggs and ovarian follicular development. BMC Res Notes. 2012 Jan 23;5:57.

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The expression of 14-3-3 proteins in the vulvar squamous epithelium. Representative samples stained using immunohistochemistry are shown. The expression of the 14-3-3 proteins (β, γ, ζ, ε, η and τ) in normal vulvar squamous epithelium (A–F). The increased cytoplasmic expression of 14-3-3β, γ, ζ, ε and η (G–K) and the reduced nuclear expression of 14-3-3τ (L) in vulvar carcinomas.

From: Wang Z, Nesland JM, Suo Z, Trope CG, Holm R (2011) The Prognostic Value of 14-3-3 Isoforms in Vulvar Squamous Cell Carcinoma Cases: 14-3-3ß and e Are Independent Prognostic Factors for These Tumors. PLoS ONE 6(9): e24843.

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The 14-3-3η protein accumulates at the metaphase II spindle of the mouse egg matured in vivo and in vitro. Cells were fixed, permeabilized and immunolabeled for confocal double immunofluorescence using one of two primary antibodies against the 14-3-3η protein (red), an antibody to a-tubulin (green) and counterstained with Hoechst 33342 (blue) to visualize DNA. (A-D) A representative in vivo-matured and ovulated egg labeled with a goat antibody that recognizes the C-terminal end of the 14-3-3η protein (C). (E-H) A representative in vitro-matured egg cell that was held in prophase I arrest for 24 hours, released from the arrest and examined at 13 hours with a rabbit antibody recognizing the N-terminal end of the 14-3-3? protein (G). PB, First Polar Body. The merged image is an overlay of immunofluorescence images from the three channels. Scale bars represent 10 µm.

From: De S, Kline D. Evidence for the requirement of 14-3-3η (YWHAH) in meiotic spindle assembly during mouse oocyte maturation. BMC Dev Biol. 2013 Apr 1;13:10.

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The 14-3-3η protein accumulates and co-localizes with α-tubulin in MI and MII meiotic spindles during oocyte maturation. Representative cells were fixed, permeabilized and immunolabeled for confocal microscopy during oocyte in vitro maturation at the times indicated. The left column shows the merged images of a-tubulin (green) and the counterstain Hoechst 33342 (blue) to visualize the DNA. The right column shows each corresponding image of the 14-3-3η protein (red). (A-H) Paired confocal images at a single confocal plane of the spindle region of representative cells at the times indicated. (I-L) Representative egg imaged at the plane of metaphase II spindle (I,J) and at a different confocal plane to show the first polar body (K,L) attached to the same egg cell. Scale bars represent 10 µm.

From: De S, Kline D. Evidence for the requirement of 14-3-3η (YWHAH) in meiotic spindle assembly during mouse oocyte maturation. BMC Dev Biol. 2013 Apr 1;13:10.

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The 14-3-3η protein interacts directly with α-tubulin. (A-C) A single confocal section through the region of the meiotic spindle of a representative in vitro-matured egg. Sites of protein 14-3-3η and a-tubulin are indicated by the green fluorescent spots (B). The cell was counterstained with Hoechst 33342 (blue) to visualize DNA (A) and the merged image is shown (C). (D) The non-confocal, brightfield image of this cell and spindle (white box). (E) A compressed stack of seven consecutive confocal scans performed at 2 µm intervals from the bottom of the spindle to its top. A marked accumulation of the in situ PLA sites of interaction of 14-3-3η with α-tubulin was observed at the meiotic spindles in eggs and along the egg cortices about the spindles (highlighted by the white box). (F-H) A representative control egg processed simultaneously for in situ PLA by identical procedure, in absence of the primary antibodies for 14-3-3η and α-tubulin. (F) The non-confocal, brightfield image of the cell and its spindle (white box). (G) A single, confocal fluorescence scan through the spindle region of the egg cell showing no in situ PLA fluorescent reaction spot. (H) A compressed stack of all confocal scans from the bottom of the egg cell to its top, showing complete absence of in situ PLA fluorescent reaction spots. Background fluorescence was minimal (G, H). Scale bars represent 10 µm.

From: De S, Kline D. Evidence for the requirement of 14-3-3η (YWHAH) in meiotic spindle assembly during mouse oocyte maturation. BMC Dev Biol. 2013 Apr 1;13:10.

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Microinjection of a morpholino against 14-3-3η causes absence or deformation of meiotic spindles in cells matured in vitro. Oocytes were injected with 0.1 mM 14-3-3η morpholino, held for 24 hours in prophase I arrest, released from the arrest for 13 hours, fixed, permeabilized and immunolabeled for confocal immunofluorescence with an antibody to &alpha-tubulin (green), the antibody to 14-3-3η protein (red), and counterstained with Hoechst 33342 (blue) to visualize the DNA. The panel on the far right is the merged overlay of immunofluorescence images from all three channels. (A-D and E-H) The upper two rows (cells 1 and 2) are images of two representative cells that have clumped DNA, no spindle and no 14-3-3η accumulation. (I-L, M-P, and Q-T) The lower 3 rows (cells 1–3) are representative images of cells that have deformed spindles, disorganized DNA and no accumulation of 14-3-3η at the spindle region. None of these cells injected with the 14-3-3η morpholino formed a first polar body, indicating that the disrupted spindles shown here are MI spindles. Scale bars represent 10 µm.

From: De S, Kline D. Evidence for the requirement of 14-3-3η (YWHAH) in meiotic spindle assembly during mouse oocyte maturation. BMC Dev Biol. 2013 Apr 1;13:10.

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Representative control eggs matured in vitro from injected oocytes, showing normal, bipolar meiotic spindles. (A-D) Images of an oocyte injected with 0.01 mM of the inverted 14-3-3η morpholino. (E-H) Images of an oocyte injected with 0.01 mM the 14-3-3γ morpholino. (I-L) Images of an oocyte injected with 10 pL of deionized water. In all cases these oocytes, representative of others (see Figure 4) both injected and uninjected, were held for 24 hours in prophase arrest, released from arrest for13 hours, fixed, permeabilized and immunolabeled for confocal immunofluorescence with an antibody to α-tubulin (green), the antibody to 14-3-3γ protein (red), and counterstained with Hoechst 3342 (blue) to visualize DNA. The panel at the far right is the merged overlay of immunofluorescence images from all three channels. In all cases, the spindle appears normal and 14-3-3η accumulates at the spindle. PB, first polar body. MO, morpholino oligonucleotide. Scale bars represent 10 µm..

From: De S, Kline D. Evidence for the requirement of 14-3-3η (YWHAH) in meiotic spindle assembly during mouse oocyte maturation. BMC Dev Biol. 2013 Apr 1;13:10.

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  • Rabbit anti 14-3-3 eta
(Rated 0.0 out of 5 based on 0 customer reviews)
  • Product Type
    Polyclonal Antibody
  • Isotype
    Polyclonal IgG
1 Formats Available
    Product CodeApplicationsDatasheetMSDSPack SizeList PriceQuantity
    AHP1046P *, WBdatasheet pdfdatasheet pdf0.1 ml
    AHP1046
    Summary
    Secondary Antibodies
    Negative Isotype Controls
    Useful Reagents
    Positive Controls
    Histology Controls
    More Images
    References
    Reviews
    -
    • Rabbit anti 14-3-3η antibody recognizes 14-3-3η in all mammals. It is a member of the 14-3-3 family which consists of 30 kDa proteins that are involved in multiple protein kinase signalling pathways, regulation of cell cycle progression, cytoskeletal structure, transcription, intracellular trafficking and targeting. Protein interactions with 14-3-3 show distinct preference for it’s different isoforms and are regulated by phosphorylation of both 14-3-3 and the bound protein.

      14-3-3η is expressed principally in the brain although it is expressed at low levels in other tissues. The protein binds alpha-synuclein, forming part of lewy bodies in parkinsons disease and has been linked to early-onset schizophrenia. The eta isoform binds both gremlin 1, which is overexpressed in human cancers, and the DAL-1/Protein 4.1B tumour suppressor. Furthermore the glucocortcoid receptor is positively regulated by 14-3-3- eta.

      Rabbit anti 14-3-3η antibody may not react with recombinant proteins that are not N-acetylated.
    • Intended Use
    • Target Species
      Sheep
    • Species Cross-Reactivity
      Target SpeciesCross Reactivity
      Mouseyes
      Humanyes
      Bovineyes
      Chickenyes
      Rabbityes
      Ratyes
      MammalsExpected from Sequence
      N.B. Antibody reactivity and working conditions may vary between species.
    • Product Form
      Serum - liquid
    • Reconstitution
    • Preparation
    • Antiserum Preparation
      Antisera to sheep 14-3-3 eta were raised by repeated immunisations of rabbit with highly purified antigen.
    • Preservative Stabilisers
      0.09%Sodium Azide (NaN3)
    • Immunogen
      Synthetic peptide corresponding to acetylated N-terminal sequence of sheep 14-3-3 eta.

      Peptide sequence: Ac.GDREQLLQRARC
    • Purity
    • Approx. Protein Concentrations
    • Reagents In The Kit
    • Preparing The Antibody
    • Test Principle
    • Buffer Solution
    • Storage
      Store at +4oC or at -20oC if preferred.
      Storage in frost-free freezers is not recommended.
      This product should be stored undiluted. Avoid repeated freezing and thawing as this may denature the antibody. Should this product contain a precipitate we recommend microcentrifugation before use.
    • Shelf Life
      18 months from date of despatch.
    • GO Terms
      regulation of synaptic plasticity
      glucocorticoid receptor signaling pathway
      insulin-like growth factor receptor binding
      transcription activator activity
      enzyme binding
      negative regulation of dendrite morphogenesis
      glucocorticoid receptor binding
      protein domain specific binding
      intracellular protein transport
      cytoplasm
      glucocorticoid catabolic process
    • UniProt
    • Entrez Gene
    • Acknowledgements
    • Regulatory
      For research purposes only
    • This product has been reported to work in the following applications. This information is derived from testing within our laboratories, peer-reviewed publications or personal communications from the originators. Please refer to references indicated for further information. For general protocol recommendations, please visit the antibody protocols page.

    • Application NameYesNoMin DilutionMax Dilution
      Western Blotting1/3000
      Immunohistology - Paraffin(1)1/3200
      (1)
      This product requires antigen retrieval using heat treatment prior to staining of paraffin sections.

    • Where this product has not been tested for use in a particular technique this does not necessarily exclude its use in such procedures. Suggested working dilutions are given as a guide only. It is recommended that the user titrates the product for use in their own system using appropriate negative/positive controls.
    • Technical Advice
    • Recommended Protocol
    • ELISA
    • Immunohistology
    • Histology Positive Control Tissue
      Normal brain tissue
    • Immunofluorescence
    • Western Blotting
      AHP1046 detects a band of approximately 30kDa in HEK293 cell lysates.
    • Instructions For Use

    Additional 14-3-3 eta Antibody Formats

    Formats Applications Sizes available
    14-3-3 eta Antibody : Serum P *, WB 0.1 ml
    • Copyright © 2016 Bio-Rad

    Recommended Secondary Antibody

      DescriptionProduct CodePack SizeApplicationsList PriceQuantity
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      STAR121B
      Sheep anti Rabbit IgG:Biotin2AB02B1 mlC, E, P, WB
      2AB02B
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      STAR36D649GA
      Sheep anti Rabbit IgG:Dylight®800STAR36D800GA0.1 mgF, IF, WB
      STAR36D800GA
      Goat anti Rabbit IgG (Fc):FITCSTAR121F1 mgF
      STAR121F
      Sheep anti Rabbit IgG:FITCSTAR34B1 mgC, F
      STAR34B
      Goat anti Rabbit IgG (Fc):HRPSTAR121P1 mgE, WB
      STAR121P
      Goat anti Rabbit IgG (H/L):HRPSTAR124P1 mgC, E, WB
      STAR124P
      Sheep anti Rabbit IgG:HRP (Multi Adsorbed)STAR541 mgC, E, WB
      STAR54
      Sheep anti Rabbit IgG:RPESTAR35A1 mlF
      STAR35A

      Recommended Negative Isotype Control

        Useful Reagents

          DescriptionProduct CodePack SizeApplicationsList PriceQuantity
          TidyBlot Western Blot Detection Reagent:HRPSTAR209P0.5 mlWB*
          STAR209P
          Rabbit anti 14-3-3 Isoform PanelPAN01725 µl X 7E *, P *, WB*
          PAN017

          Recommended Positive Controls

            Histology Controls

              Normal brain tissue

              References

              Further Reading

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