Anti-Sarcomeric Alpha Actinin antibody [EP2529Y]

9 product images

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Sarcomeric Alpha Actinin antibody [EP2529Y] (ab68167)

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse cardiac muscle tissue labelling Sarcomeric Alpha Actinin with purified ab68167 at 1/100. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. Goat Anti-Rabbit IgG H&L (HRP) ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

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  Western blot - Anti-Sarcomeric Alpha Actinin antibody [EP2529Y] (ab68167)

  Blocking and dilution buffer: 5% NFDM /TBST.

  All lanes: Western blot - Anti-Sarcomeric Alpha Actinin antibody [EP2529Y] (ab68167) at 1/20000 dilution

  All lanes: Human fetal heart tissue lysate at 10 µg

  Secondary

  All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution

  Predicted band size: 103 kDa

  Observed band size: 103 kDa

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  Immunoprecipitation - Anti-Sarcomeric Alpha Actinin antibody [EP2529Y] (ab68167)

  ab68167 (purified) at 1/20 immunoprecipitating Sarcomeric Alpha Actinin in HepG2 whole cell lysate.
  

  Lane 1 (input): HepG2 whole cell lysate (10μg)

  Lane 2 (+): ab68167 + HepG2 whole cell lysate.

  Lane 3 (-): Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab68167 in HepG2 whole cell lysate.

  For western blotting, VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366), was used for detection at 1/10,000 dilution.

  Blocking buffer and concentration: 5% NFDM/TBST.

  Diluting buffer and concentration: 5% NFDM /TBST.

  All lanes: Immunoprecipitation - Anti-Sarcomeric Alpha Actinin antibody [EP2529Y] (ab68167) at 1/20 dilution

  Lane 1: HepG2 whole cell lysate at 10 µg

  Lane 2: ab68167 + HepG2 whole cell lysate at 10 µg

  Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab68167 in HepG2 whole cell lysate

  Predicted band size: 103 kDa

  Observed band size: 103 kDa

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Sarcomeric Alpha Actinin antibody [EP2529Y] (ab68167)

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human muscle tissue labelling Sarcomeric Alpha Actin with unpurified ab68167 at a dilution of 1/100.
  

  Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Sarcomeric Alpha Actinin antibody [EP2529Y] (ab68167)

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human cardiac muscle tissue labelling Sarcomeric Alpha Actinin with purified ab68167 at 1/100. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. Goat Anti-Rabbit IgG H&L (HRP) ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

• 

  Western blot - Anti-Sarcomeric Alpha Actinin antibody [EP2529Y] (ab68167)

  All lanes: Western blot - Anti-Sarcomeric Alpha Actinin antibody [EP2529Y] (ab68167) at 1/20000 dilution

  Lane 1: Human skeletal muscle lysate at 10 µg

  Lane 2: Rat heart lysate at 10 µg

  Lane 3: Human heart lysate at 10 µg

  Secondary

  All lanes: HRP-conjugated goat anti-rabbit IgG at 1/2000 dilution

  Predicted band size: 103 kDa

  Observed band size: 103 kDa

• 

  Western blot - Anti-Sarcomeric Alpha Actinin antibody [EP2529Y] (ab68167)

  Blocking and dilution buffer: 5% NFDM /TBST.

  All lanes: Western blot - Anti-Sarcomeric Alpha Actinin antibody [EP2529Y] (ab68167) at 1/20000 dilution

  All lanes: Mouse heart tissue lysate at 10 µg

  Secondary

  All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution

  Predicted band size: 103 kDa

  Observed band size: 103 kDa

• 

  Western blot - Anti-Sarcomeric Alpha Actinin antibody [EP2529Y] (ab68167)

  Blocking and dilution buffer: 5% NFDM /TBST.

  All lanes: Western blot - Anti-Sarcomeric Alpha Actinin antibody [EP2529Y] (ab68167) at 1/10000 dilution

  Lane 1: Mouse brain tissue lysate at 10 µg

  Lane 2: Rat brain tissue lysate at 10 µg

  Secondary

  All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution

  Predicted band size: 103 kDa

  Observed band size: 103 kDa

• 

  Image collected and cropped by CiteAb under a CC-BY license from the publication

  Western blot - Anti-Sarcomeric Alpha Actinin antibody [EP2529Y] (ab68167)

  Sarcomeric Alpha Actinin western blot using anti-Sarcomeric Alpha Actinin antibody [EP2529Y] ab68167. Publication image and figure legend from Hodges, J. L., Vilchez, S. M., et al., 2014, PLoS One, PubMed 25007055.

  
  

  ab68167 was used in this publication in western blot. This may not be the same as the application(s) guaranteed by Abcam. For a full list of applications guaranteed by Abcam for ab68167 please see the product overview.

  α-actinin-2 localizes to post-synaptic sites in dendritic spines on hippocampal neurons.A) An anti-α-actinin antibody (ab68167) recognizes α-actinin-2 and not α-actinin-1 or α-actinin-4. CHO-K1 cells were transfected with human α-actinin-1-GFP, α-actinin-2-GFP, or α-actinin-4-GFP. Cells were lysed and immunoblotted for α-actinin-2 and GFP. B) α-Actinin-2 is enriched in hippocampal neurons but not in glia cells or COS-7 cells, which lacks α-actinin-2. Cells were lysed and immunoblotted for α-actinin-2. Actin is the loading control. C) α-Actinin-2 localizes to dendritic spines. Hippocampal neurons were transfected at DIV 17 with GFP (green), and fixed, and immunostained for endogenous α-actinin-2 (magenta) at DIV 21. D) α-Actinin-2 co-localizes with post-synaptic markers, but not with a pre-synaptic marker. Hippocampal neurons were fixed at DIV 16 or 21 and immunostained for endogenous α-actinin-2 (green) and either endogenous synaptophysin, PSD-95, or the NR1 subunit of the NMDA receptor (magenta). E–G) The siRNA is specific for α-actinin-2. Hippocampal neurons were co-transfected at DIV 17 with GFP and either a control empty vector (pSUPER), or a vector containing siRNA against α-actinin-2 (pSUPER-α-actinin-2), or the α-actinin-2 siRNA-containing vector plus a α-actinin-2 vector conferring resistance to RNAi (pSUPER-α-actinin-2+ α-actinin-2-SS). The cells were fixed at DIV 21 and immunostained for endogenous α-actinin-2. Arrows point to the neurons co-expressing GFP and its immunostaining for α-actinin-2. For each condition (55 control cells and 46 α-actinin-2 knockdown cells), the integrated density of the cell body and dendrites were measured from the transfected neuron and adjacent untransfected neuron of the same image and the percent change was plotted, F. Error bars represent SEM. p-values were derived using the paired t-test. G) CHO-K1 cells were co-transfected with GFP, pSUPER or pSUPER-α-actinin-2, plus either α-actinin-2-Flag or α-actinin-2-SS-Flag. Transfection efficiency was close to 100% as >95% of the cells in each condition exhibited GFP fluorescence (data not shown). Cells were lysed 72 hours after transfection and immunoblotted for α-actinin-2 and α-actinin-4. Tubulin is the loading control.