14-3-3 Isoform Panel

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14-3-3 Isoform Panel 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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14-3-3 Isoform Panel gallery image 2

Published customer image:
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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14-3-3 Isoform Panel gallery image 3

Published customer image:
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). 14-3-3η accumulates, in part, in the meiotic spindle in eggs as shown by simultaneous labeling with 14-3-3η (Q) and tubulin (R) antibodies. These sequential scans are merged (14-3-3η + 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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14-3-3 Isoform Panel gallery image 4

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Representative immunohistochemistry images of 14-3-3 β in the different stages of follicular development in ovarian sections. Brown staining represents 14-3-3 β against regions counterstained blue with hematoxylin. (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), the more intense stain along the zona pellucida of the oocyte (C, yellow arrow), and the more intense staining in cells lining the antral cavity (D and 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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14-3-3 Isoform Panel gallery image 5

Published customer image:
Representative immunohistochemistry images of 14-3-3 γ 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 (D and 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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14-3-3 Isoform Panel gallery image 6

Published customer image:
Representative immunohistochemistry images of 14-3-3 ε 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), the more intense stain along the zona pellucida of the oocyte (C, yellow arrow), 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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14-3-3 Isoform Panel gallery image 7

Published customer image:
Representative immunohistochemistry images of 14-3-3 ζ 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 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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14-3-3 Isoform Panel gallery image 8

Published customer image:
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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14-3-3 Isoform Panel gallery image 9

Published customer image:
Representative immunohistochemistry images of 14-3-3 τ 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), the more intense stain along the zona pellucida of the oocyte (C, yellow arrow), 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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14-3-3 Isoform Panel gallery image 10

Published customer image:
Representative immunohistochemistry images of 14-3-3 σ 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), the more intense stain along the zona pellucida of the oocyte (C, yellow arrow), and the more intense staining in cells lining the antral cavity (D and 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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14-3-3 Isoform Panel gallery image 11

Published customer image:
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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14-3-3 Isoform Panel gallery image 12

Published customer image:
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 β. (C,D) 14-3-3 γ. (E,F) 14-3-3 ε. (G,H) 14-3-3 ζ. (I,J) 14-3-3 η. (K,L) 14-3-3 τ. (M,N) 14-3-3 ε. (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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14-3-3 Isoform Panel gallery image 13

Published customer image:
Representative control immunohistochemistry images for the different 14-3-3 protein isoforms in tissue sections. (A) Example of a positive control brain section obtained for all 14-3-3 isoforms except for 14-3-3 σ. (B) Skin positive control for 14-3-3 σ. (C) Example of a negative control section with no primary antibody obtained for all isoforms. (D) Example of a negative control with rabbit serum in place of primary antibody obtained for all isoforms. 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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  • Rabbit anti 14-3-3 Isoform Panel
(Rated 0.0 out of 5 based on 0 customer reviews)
  • Product Type
    Kits
  • Isotype
    Polyclonal IgG
1 Formats Available
    Product CodeApplicationsDatasheetMSDSPack SizeList PriceQuantity
    PAN017E *, P *, WB*datasheet pdfdatasheet pdf25 µl X 7
    PAN017
    Summary
    Secondary Antibodies
    Negative Isotype Controls
    Useful Reagents
    Positive Controls
    Histology Controls
    More Images
    References
    Reviews
    -
    • Rabbit anti 14-3-3 Isoform antibody Panel provides the user with a sample panel of the 7 mammalian isoforms of 14-3-3 protein, each component part recognising the acetylated N-terminal region of the relevant isoform. Note: may not react with recombinant proteins that are not N-acetylated.

      14-3-3 is a member of the 14-3-3 family, consisting of acidic 30 kDa proteins 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 preferences for the different isoforms, and are regulated by phosphorylation of both 14-3-3 and the bound protein.
    • Intended Use
    • Target Species
      Sheep
    • Product Form
      Serum - liquid
    • Reconstitution
    • Preparation
    • Antiserum Preparation
      Antisera to the relevant sheep 14-3-3 isoform were raised by repeated immunisations of rabbits with highly purified antigen.
    • Preservative Stabilisers
      0.09%Sodium Azide (NaN3)
    • Immunogen
      Synthetic peptides corresponding to acetylated N-terminal sequences of sheep 14-3-3 beta, epsilon, eta, gamma, sigma, tau/theta, zeta accordingly.
    • Purity
    • Approx. Protein Concentrations
    • Reagents In The Kit
      PAN017 is made up of 25ul of each of the following component parts, which are also available individually in 0.1ml size.

      1. 14-3-3 beta (AHP1044) - plays a role in the pathogenesis of systemic lupus erythematosus and is involved in negative regulation of p90 ribosomal S6 kinase activity and tuberous sclerosis tumour suppressor proteins. Binds IGF-1 receptor, RAF1 and CDC25 phosphatases.

      Cross-reactivity: Human, Rat, Mouse, Rabbit, Bovine N.B. Antibody reactivity and working conditions may vary between species.

      Immunohistology: requires antigen retrieval using heat treatment prior to staining of paraffin sections. Normal brain tissue may be used as a positive control.

      2. 14-3-3 epsilon (AHP1048) is expressed in all mammals and is a potential suicide susceptibility gene. It is deleted in small cell lung cancers disrupting the G(2) checkpoint. During apoptosis, cleavage by caspase-3 increases the affinity between bad and Bcl-x(L). When bound by ERK, 14-3-3 epsilon is thought to deactivate heat shock factor 1 in cells recovering from heat shock.

      Cross-reactivity: Human, Rat, Mouse, Rabbit, Bovine, Chicken N.B. Antibody reactivity and working conditions may vary between species.

      Immunohistology: does not require antigen retrieval using heat treatment prior to staining of paraffin sections. Normal brain tissue may be used as a positive control.

      3. 14-3-3 eta (AHP1046) is expressed principally in the brain and at low levels in other tissues. It binds alpha-synuclein, forming part of Lewy bodies in Parkinson’s disease and has been linked to early-onset schizophrenia. It 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.

      Cross-reactivity: Human, Rat, Mouse, Rabbit, Bovine, Chicken N.B. Antibody reactivity and working conditions may vary between species.

      Immunohistology: does not require antigen retrieval using heat treatment prior to staining of paraffin sections. Normal brain tissue may be used as a positive control.

      4. 14-3-3 gamma (AHP1047) is thought to play an important role in muscle, being induced by growth factors in human vascular smooth muscle cells, and being expressed in skeletal and heart muscles. It binds amyloid beta-protein precursor intracellular domain fragment (AICD) and FE65, facilitating FE65-dependent gene transactivation. Binding of 14-3-3 gamma to glial fibrillary acidic protein is important in the regulation of glial filaments. It binds and may regulate CDK11 during the cell cycle and apoptosis. The level of this isoform is lowered in foetal Down’s syndrome brains.

      Cross-reactivity: Human, Rat, Mouse, Bovine N.B. Antibody reactivity and working conditions may vary between species.

      Immunohistology: requires antigen retrieval using heat treatment prior to staining of paraffin sections. Normal brain tissue may be used as a positive control.

      5. 14-3-3 sigma (AHP1050) is expressed mostly in epithelial cells and has been associated with cancers including, nasopharyngeal, endometrial adenocarcinoma, prostate, epithelial ovarian, colorectal, adenoid cystic, papillary and breast.

      14-3-3 sigma binds c-fos activating the MAPK pathway and regulates mitosis through binding keratin filaments and sequestering cyclin B1 and cdc2. Sequestration of Bax by 14-3-3 sigma inhibits apoptosis whereas sequestration by CARP-1 induces apoptosis. p73-dependent induction of 14-3-3 sigma regulates chemo sensitivity in breast cancers with p53 mutations. Along with RACK1, 14-3-3 sigma targets DeltaNp63alpha for proteasome degradation, in reaction to DNA damage.

      Cross-reactivity: Human, Rat, Mouse, Rabbit, Bovine, Chicken N.B. Antibody reactivity and working conditions may vary between species.

      6. 14-3-3 tau/theta (AHP1051) is primarily expressed in T-cells and occurs at lower levels in the brain, heart, pancreas placenta and kidneys. It plays a role in cell cycle progression via interaction with p27(Kip1) and, along with COPI, binds the GB1 RSR sequence involved in protein trafficking. 14-3-3 tau/theta forms a complex with Hsp60 and cellular prion protein which may be involved in prion diseases. It binds BCR/Abl, bacterial effector protein Tir, yes-associated protein and the FSH receptor. Elevated levels of 14-3-3 tau/theta are observed in amyotrophic lateral sclerosis.

      Cross-reactivity: Human, Rat, Mouse, Rabbit, Bovine, Chicken N.B.

      Antibody reactivity and working conditions may vary between species. Immunohistology: does not require antigen retrieval using heat treatment prior to staining of paraffin sections. Normal tonsil tissue may be used as a positive control.

      7. 14-3-3 zeta (AHP1052) is an adaptor protein implicated as a regulator of an array of signalling pathways. It binds to and modulates the activity of a large number of protein partners, usually by recognition of a phosphoserine or phosphothreonine motif.

      Cross-reactivity: Human, Rat, Mouse, Bovine, Chicken N.B. Antibody reactivity and working conditions may vary between species.

      Immunohistology: does not require antigen retrieval using heat treatment prior to staining of paraffin sections. Normal brain tissue may be used as a positive control.
    • 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.
    • 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
      ELISA(1)
      Immunohistology - Paraffin(2)
      Western Blotting(3)
      (1)
      All isoforms except eta and sigma
      (2)
      All isoforms except sigma
      (3)
      All isoforms

    • 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
    • Immunofluorescence
    • Western Blotting
      All PAN017 component parts detect a band of approximately 30kDa in HEK293 cell lysates.
    • Instructions For Use

    Additional 14-3-3 Isoform Panel Formats

    Formats Applications Sizes available
    14-3-3 Isoform Panel : Serum E *, P *, WB* 25 µl X 7
    • Copyright © 2016 Bio-Rad

    Recommended Secondary Antibody

      Recommended Negative Isotype Control

        Useful Reagents

          DescriptionProduct CodePack SizeApplicationsList PriceQuantity
          Rabbit anti 14-3-3 BetaAHP10440.1 mlE, P *, WB
          AHP1044
          Rabbit anti 14-3-3 Epsilon (C-Terminal)AHP10450.1 mlE, P, WB
          AHP1045
          Rabbit anti 14-3-3 etaAHP10460.1 mlP *, WB
          AHP1046
          Rabbit anti 14-3-3 GammaAHP10470.1 mlE, P *, WB
          AHP1047
          Rabbit anti 14-3-3 Epsilon (N-Terminal)AHP10480.1 mlE, P *, WB
          AHP1048
          Rabbit anti 14-3-3 SigmaAHP10500.1 mlWB
          AHP1050
          Rabbit anti 14-3-3 tau/thetaAHP10510.1 mlE, P *, WB
          AHP1051
          Rabbit anti 14-3-3 Zeta/DeltaAHP10520.1 mlE, P *, WB
          AHP1052

          Recommended Positive Controls

            Histology Controls

              References

              Further Reading

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