Anti-CD40 antibody [EPR20540] - Low endotoxin, Azide free

5 product images

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  Western blot - Anti-CD40 antibody [EPR20540] - Low endotoxin, Azide free (ab223546)

  This data was developed using the same antibody clone in a different buffer formulation (Anti-CD40 antibody [EPR20540] ab213205).
  

  Lanes 1 - 3: Merged signal (red and green). Green - Anti-CD40 antibody [EPR20540] ab213205 observed at 42 kDa. Red - loading control, Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (Mouse anti-Alpha Tubulin [DM1A] observed at 55kDa.

  Anti-CD40 antibody [EPR20540] ab213205 was shown to react with CD40 in U-2 OS wild-type cells in Western blot. Loss of signal was observed when CD40 knockout sample was used. U-2 OS wild-type and CD40 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% Milk in TBS-T (0.1% Tween®) before incubation with Anti-CD40 antibody [EPR20540] ab213205 and Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (Mouse anti-Alpha Tubulin [DM1A] overnight at 4°C at a 1 in 2000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

  All lanes: Western blot - Anti-CD40 antibody [EPR20540] (Anti-CD40 antibody [EPR20540] ab213205) at 1/2000 dilution

  Lane 1: Wild-type U-2 OS whole cell lysate at 20 µg

  Lane 2: CD40 knockout U-2 OS whole cell lysate at 20 µg

  Lane 3: Raji (Human Burkitt's lymphoma cell line) whole cell lysate at 20 µg

  Performed under reducing conditions.

  Predicted band size: 30 kDa

  Observed band size: 42 kDa

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  This image is courtesy of the Single Cell & Imaging Mass Cytometry Analysis Platform, Goodman Cancer Research Centre, McGill University

  Mass Cytometry - Anti-CD40 antibody [EPR20540] - Low endotoxin, Azide free (ab223546)

  Imaging Mass Cytometry™ (IMC™) image of human tonsil tissue stained with Anti-CD40 antibody [EPR20540]. ab223546 (carrier-free antibody, purified) was metal-conjugated using a Maxpar® Antibody Labeling Kit from Fluidigm. Immunostaining was performed according to Fluidigm’s protocols. Briefly, slides were subject to deparaffinization and heat-induced epitope retrieval, followed by overnight incubation at 4°C with an antibody cocktail containing metal-tagged antibodies in blocking buffer. Slides were subsequently washed with 0.2% Triton-X and 1x PBS, counterstained with Cell-ID™ Intercalator-Ir diluted at 1/400 in 1x PBS for 30 min at room temperature, rinsed for 5 min with distilled H2O, and air-dried prior to IMC™ acquisition. IMC™ acquisition was performed using the Fluidigm Hyperion™ Imaging System.
  Imaging Mass Cytometry™, IMC™, Cell-ID™, Hyperion™ and Maxpar® are trademarks of Fluidigm Canada

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD40 antibody [EPR20540] - Low endotoxin, Azide free (ab223546)

  This IHC data was generated using the same anti-CD40 antibody clone [EPR20540] in a different buffer formulation (cat# Anti-CD40 antibody [EPR20540] ab213205).
  

  Immunohistochemical analysis of paraffin-embedded human tonsil tissue labeling CD40 with Anti-CD40 antibody [EPR20540] ab213205 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/500 dilution.

  Membranous and cytoplasmic staining on germinal center of human tonsil is observed [PMID: 10360965] [PMID: 7507299] [PMID:24452203].

  Counter stained with Hematoxylin.

  Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/500 dilution.

  Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD40 antibody [EPR20540] - Low endotoxin, Azide free (ab223546)

  Immunohistochemical analysis of paraffin-embedded human large B cell lymphoma tissue labeling CD40 with Anti-CD40 antibody [EPR20540] ab213205 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/500 dilution.
  

  Membranous staining on human large B cell lymphoma is observed [PMID: 7507299].

  Counter stained with Hematoxylin.

  Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/500 dilution.

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-CD40 antibody [EPR20540] ab213205).

  Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD40 antibody [EPR20540] - Low endotoxin, Azide free (ab223546)

  Tissue Microarrays stained for "Anti-CD40 antibody [EPR20540]" using "Anti-CD40 antibody [EPR20540] ab213205"in immunohistochemical analysis. This table provides a detailed overview of positive (tick mark) and negative (cross mark) staining per sample type tested. The sections were pre-treated using Heat mediated antigen retrieval using Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) ab93684 (Tris/EDTA buffer, pH 9.0). The sections were incubated with Anti-CD40 antibody [EPR20540] ab213205 at +4°C overnight followed by Goat Anti-Rabbit IgG H&L (HRP) Goat Anti-Rabbit IgG H&L (HRP) ab97051 at 1/500.