Anti-EGFR antibody [EP38Y] - BSA and Azide free

11 product images

• 

  Immunoprecipitation - Anti-EGFR antibody [EP38Y] - BSA and Azide free (ab272293)

  Anti-EGFR antibody [EP38Y] ab52894 (purified) at 1:20 dilution (0.5 μg) immunoprecipitating EGFR in HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate.
  
  Lane 1 (input): HeLa whole cell lysate 10 μg
  
  Lane 2 (+): Anti-EGFR antibody [EP38Y] ab52894 in HeLa whole cell lysate
  
  Lane 3 (-): Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of Anti-EGFR antibody [EP38Y] ab52894 in HeLa whole cell lysate
  
  For western blotting, VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) was used for detection at 1:1000 dilution.
  
  Blocking and diluting buffer: 5% NFDM/TBST.
  

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-EGFR antibody [EP38Y] ab52894).

  All lanes: Immunoprecipitation - Anti-EGFR antibody [EP38Y] (Anti-EGFR antibody [EP38Y] ab52894)

  Predicted band size: 134 kDa

  Observed band size: 175 kDa

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EGFR antibody [EP38Y] - BSA and Azide free (ab272293)

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human cervical carcinoma tissue sections labeling EGFR with purified Anti-EGFR antibody [EP38Y] ab52894 at 1:100 dilution (0.95 μg/ml).
  

  Heat mediated antigen retrieval was performed using EDTA buffer, pH 9.0. Tissue was counterstained with hematoxylin. Goat Anti-Rabbit IgG H&L (HRP) ab97051 Goat Anti-Rabbit IgG H&L (HRP) secondary antibody was used at 1:500 dilution. PBS instead of the primary antibody was used as the negative control.

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-EGFR antibody [EP38Y] ab52894).

• 

  Western blot - Anti-EGFR antibody [EP38Y] - BSA and Azide free (ab272293)

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-EGFR antibody [EP38Y] ab52894).
  

  False colour image of Western blot: Anti-EGFR antibody [EP38Y] staining at 1/1000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, Anti-EGFR antibody [EP38Y] ab52894 was shown to bind specifically to EGFR. A band was observed at 160 kDa in wild-type HCT 116 cell lysates with no signal observed at this size in Egfr CRISPR-Cas9 edited cell line Human EGFR knockout HCT116 cell line ab281597 (CRISPR-Cas9 edited cell lysate ab282949). The band observed in the CRISPR-Cas9 edited lysate lane below 160 kDa is likely to represent a truncated form of EGFR. This has not been investigated further and the functional properties of the gene product have not been determined. To generate this image, wild-type and Egfr CRISPR-Cas9 edited HCT 116 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4°C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were 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) at 1/20000 dilution.

  Lane 1: Wild-type HCT 116 cell lysate at 20 µg

  Lane 2: EGFR CRISPR-Cas9 edited HCT 116 cell lysate at 20 µg

  Lane 3: Caco-2 cell lysate at 20 µg

  Performed under reducing conditions.

  Predicted band size: 134 kDa

  Observed band size: 160 kDa

• 

  Flow Cytometry (Intracellular) - Anti-EGFR antibody [EP38Y] - BSA and Azide free (ab272293)

  Intracellular Flow Cytometry analysis of A431 (Human epidermoid carcinoma epithelial cell) cells labeling EGFR with purified Anti-EGFR antibody [EP38Y] ab52894.
  

  Cells were fixed with 4% paraformaldehyde (10 mins) and permeabilized with 90% methanol for 30 mins. Then incubated in 1x PBS / 10% normal goat serum to block non-specific protein-protein interactions followed by Anti-EGFR antibody [EP38Y] ab52894 at 1/20 dilution (red) for 30 mins. A Goat anti rabbit IgG (Alexa Fluor^® 488) secondary antibody was used at 1/2000 dilution. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-EGFR antibody [EP38Y] ab52894).

• 

  Immunocytochemistry/ Immunofluorescence - Anti-EGFR antibody [EP38Y] - BSA and Azide free (ab272293)

  Immunocytochemistry/ Immunofluorescence analysis of A431 (Human epidermoid carcinoma epithelial cell) cells labeling EGFR with purified Anti-EGFR antibody [EP38Y] ab52894 at 1:250 dilution (0.4 μg/ml).
  

  Cells were fixed in 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. Cells were counterstained with Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) 1:200 (2.5 μg/ml). Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 Goat anti rabbit IgG(Alexa Fluor^® 488) was used as the secondary antibody at 1:1000 dilution. DAPI nuclear counterstain.

  PBS instead of the primary antibody was used as the secondary antibody only control.

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-EGFR antibody [EP38Y] ab52894).

• 

  OI-RD Scanning - Anti-EGFR antibody [EP38Y] - BSA and Azide free (ab272293)

  We have systematically measured KD (the equilibrium dissociation constant between the antibody and its antigen), of more than 840 recombinant antibodies to assess not only their individual KD values but also to see the average affinity of antibody.
  Based on the comparison with published literature values for mouse monoclonal antibodies, Recombinant antibodies appear to be on average 1-2 order of magnitude higher affinity.

• 

  Western blot - Anti-EGFR antibody [EP38Y] - BSA and Azide free (ab272293)

  This data was developed using Anti-EGFR antibody [EP38Y] ab52894, the same antibody clone in a different buffer formulation. Different batches of Anti-EGFR antibody [EP38Y] ab52894 were tested on HeLa (Human cervix adenocarcinoma epithelial cell) lysate at 1.0 µg/ml. 15 µg of lysate was loaded in each lane. Bands observed at 175 kDa.

  All lanes: Western blot - Anti-EGFR antibody [EP38Y] (Anti-EGFR antibody [EP38Y] ab52894)

  Predicted band size: 134 kDa

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EGFR antibody [EP38Y] - BSA and Azide free (ab272293)

  This data was developed using the same antibody clone in a different buffer formulation (Anti-EGFR antibody [EP38Y] ab52894).
  

  Tissue microarrays stained for Anti-EGFR antibody [EP38Y] using Anti-EGFR antibody [EP38Y] ab52894 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 was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins. The section was incubated with Anti-EGFR antibody [EP38Y] ab52894 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EGFR antibody [EP38Y] - BSA and Azide free (ab272293)

  This data was developed using the same antibody clone in a different buffer formulation (Anti-EGFR antibody [EP38Y] ab52894).

  Immunohistochemical analysis of formalin fixed paraffin embedded human placenta labelling EGFR with Anti-EGFR antibody [EP38Y] ab52894 at a concentration of 0.1 µg/ml. The immunostaining was performed on a Ventana DISCOVERY ULTRA (Roche Tissue Diagnostics) instrument with an OptiView DAB IHC Detection Kit. Heat mediated antigen retrieval was conducted for 32 mins at 100°C with ULTRA cell conditioning solution (CC1, pH 8.5). Anti-EGFR antibody [EP38Y] ab52894 Anti-EGFR antibody [EP38Y] was incubated at 37°C for 16 mins. Sections were counterstained with Hematoxylin II. Image inset shows absence of staining in secondary antibody only control.

• 

  Western blot - Anti-EGFR antibody [EP38Y] - BSA and Azide free (ab272293)

  Western blot: Anti-EGFR antibody [EP38Y] (Anti-EGFR antibody [EP38Y] ab52894) staining at 1/1000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, Anti-EGFR antibody [EP38Y] ab52894 was shown to bind specifically to EGFR. A band was observed at 175 kDa in wild-type A549 cell lysates with no signal observed at this size in EGFR knockout cell line. To generate this image, wild-type and EGFR knockout A549 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5% milk in TBS-0.1% Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4°C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

  Lanes 1 - 4: Western blot - Anti-EGFR antibody [EP38Y] - BSA and Azide free (ab272293)

  Lanes 1 - 4: Western blot - Human EGFR knockout A549 cell line (Human EGFR knockout A549 cell line ab286394)

  Lane 1: Wild-type A549 cell lysate at 20 µg

  Lane 2: EGFR knockout A549 cell lysate at 20 µg

  Lane 3: Wild-type HeLa cell lysate at 20 µg

  Lane 4: EGFR knockout HeLa cell lysate at 20 µg

  Developed using the ECL technique.

  Performed under reducing conditions.

  Observed band size: 175 kDa

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EGFR antibody [EP38Y] - BSA and Azide free (ab272293)

  This data was developed using the same antibody clone in a different buffer formulation (Anti-EGFR antibody [EP38Y] ab52894).

  Immunohistochemical analysis of formalin fixed paraffin embedded human placenta labelling EGFR with Anti-EGFR antibody [EP38Y] ab52894 at a concentration of 0.21 µg/ml. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument with a Bond™ Polymer Refine Detection kit. Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution 2) for 20 mins. Anti-EGFR antibody [EP38Y] ab52894 Anti-EGFR antibody [EP38Y] was incubated for 30 mins at room temperature. Sections were counterstained with Hematoxylin. Image inset shows absence of staining in secondary antibody only control.