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Monoclonal Antibodies

Anti-Talin2 [121A]

Invented at University Of Leicester
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Adapted from Praekelt U, Kopp PM, Rehm K, Linder S, Bate N, Patel B, Debrand E, Manso AM, Ross RS, Conti F, Zhang MZ, Harris RC, Zent R, Critchley DR, Monkley SJ. New isoform-specific monoclonal antibodies reveal different sub-cellular localisations for talin1 and talin2. Eur J Cell Biol. 2012 Mar;91(3):180-91. doi: 10.1016/j.ejcb.2011.12.003. Epub 2012 Feb 3. PMID: 22306379; PMCID: PMC3629562.Characterisation of isoform-specific talin monoclonal antibodies. (A) Domain structure of talin. The N-terminal talin head, which is comprised of an atypical FERM domain, is linked to the talin rod by an unstructured region (zig-zag) containing a calpain-II cleavage site. The talin rod contains 61 α-helices (grey ovals) organised into a series of amphipathic α-helical bundles, and terminates in a single helix responsible for talin dimerisation. Boxes indicate established domain boundaries. The location of the recombinant polypeptides used to generate Mabs and the position of their epitopes is shown either above (talin1) or below (talin2) the diagram. (B) Purified recombinant talin rod polypeptides (50 ng) spanning residues 489–911 (upper panel) or 2300–2542 (lower panel) were resolved by SDS-PAGE and either stained with Coomassie blue (coo) or analysed by Western blotting using talin Mabs 97H6, 93E12, 68E7, 121A as well as the commercial talin Mab 8d4 (Sigma). (C) The epitopes recognised by Mab 93E12 and 97H6 were further defined using recombinant talin1 rod polypeptides containing either residues 482–655 or residues 656–911. Mab 97H6 recognises the former while 93E12 recognises the latter. (D) Western blots of whole cell lysates from untransfected mouse embryo fibroblasts (40 μg protein) or lysates from cells expressing either GFP-talin1, GFP-talin2 or GFP alone (2.5 μg protein) probed with the talin Mabs indicated. (E) Mouse embryo fibroblasts homozygous for a conditional Tln1 allele and carrying a tamoxifen-inducible Cre recombinase were treated with 100 nM 4-hydoxy tamoxifen (4-OHT) dissolved in ethanol (+) or ethanol alone (−). After 72 h, cells were replated, and after a further 24 h analysed for talin1 expression by Western blotting using the 97H6 Mab. Vinculin was used as a loading control. (F) Heterozygous Tln2 knockout mice (+/−) were crossed, and 12.5dpc (+/−) or (−/−) embryos analysed for expression of talin1 (97H6 Mab) or talin2 (68E7 Mab) by Western blotting. (G) Lysates from NIH3T3 were immunoprecipitated with either a control IgG, the talin1-specific Mab 97H6 or the talin2-specific Mabs 68E7 and 121A, and the immunoprecipitates and whole cell lysate (WCL) analysed by Western blotting using the talin1-specific Mab 97H6, the talin2-specific Mab 68E7, the β1-integrin Mab (BD Transduction) and a vinculin Mab (Sigma). (H) Tissue lysates (TL) from mouse kidney were immunoprecipitated with either a control IgG, the talin2-specific Mab 68E7 or the talin1-specific, Mab 97H6 and immunoprecipitates analysed by Western blotting using the talin2-specific Mab 121A. The ∼90 kDa protein detected by Mab 121A in kidney TL is a C-terminal talin2 polypeptide spanning residues 1608–2543 (Debrand et al., 2009) which is not recognised by the 68E7 Mab.
Adapted from Praekelt U, Kopp PM, Rehm K, Linder S, Bate N, Patel B, Debrand E, Manso AM, Ross RS, Conti F, Zhang MZ, Harris RC, Zent R, Critchley DR, Monkley SJ. New isoform-specific monoclonal antibodies reveal different sub-cellular localisations for talin1 and talin2. Eur J Cell Biol. 2012 Mar;91(3):180-91. doi: 10.1016/j.ejcb.2011.12.003. Epub 2012 Feb 3. PMID: 22306379; PMCID: PMC3629562. Expression of talin1 and talin2 in various mouse tissues and their immuno-localisation in muscle and kidney. (A) Western blot of whole tissue lysates (40 μg protein) probed with the talin1- or talin2-specific antibodies shown. Tissues: B, brain; H, heart; S, spleen; K, kidney; Lu, lung; Li, liver; M, skeletal muscle. (B) Cryo-sections of gastrocnemius muscle were immunostained with Mab 97H6 directed against talin1 (a and b) and Mab 68E7 against talin2 (c–f). Talin1 was mostly localised to the myotendinous junction (MTJ) (arrows in a), with weaker staining of costameres (b), while talin2 staining was strong at both MTJ (arrows in c) and costameres (d). In talin2-null muscle, no staining was detected at the MTJ (arrows in e) or at costameres (f). Scale bar = 50 μm. (C) Immuno-staining of paraffin-embedded sections of wild type and talin2 null mouse kidney with antibodies to talin1 (93E12 and 97H6) and talin2 (68E7 and 121A). Staining is present in the glomerulus, tubules and interstitium in the cortex, and the tubules and interstitium in the medulla. There was no staining for talin2 in kidneys from Tln2-null mice.
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Catalogue Number 151628
Applications ELISA IHC IF IP WB
Antigen/Gene or Protein Targets Talin 2
Reactivity Human
Relevance Talin 2 is a high molecular weight protein found in a variety of tissues and cell types. Talin 2 interacts with integrins but it's full functions are not known. This antibody can be used for the specific detection of the focal adhesion protein talin 2 and does not cross react with talin 1.
Host Mouse
Immunogen Recombinant protein fragment
Positive Control Fibroblasts
Subclass IgG2b
Myeloma Used NS0
Recommended Growing Conditions RPMI 1650 + Glu + Pen + Strep + 10% FCS @ 37C + 5% CO2
Notes Suggested Dilutions: 1/100 - 1/1000
Research Area Cell Signaling & Signal Transduction

References

There are 2 reference entries for this reagent.

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References: 2 entries

Praekelt et al. 2012. Eur J Cell Biol. 91(3):180-91. PMID: 22306379.

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New isoform-specific monoclonal antibodies reveal different sub-cellular localisations for talin1 and talin2.

Europe PMC ID: 22306379

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References: 2 entries

Praekelt et al. 2012. Eur J Cell Biol. 91(3):180-91. PMID: 22306379.

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New isoform-specific monoclonal antibodies reveal different sub-cellular localisations for talin1 and talin2.

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Invented at

University Of Leicester
University Of Leicester

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