Anti-Melanoma gp100 antibody [EP4863(2)]

8 product images

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  Immunoprecipitation - Anti-Melanoma gp100 antibody [EP4863(2)] (ab137078)

  ab137078 immunoprecipitating Melanoma gp100. 10μg of cell lysate was incubated with primary antibody at a dilution of 1/500 and VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) at a dilution of 1/10000.
  

  Lane 1: B16-F0 (mouse melanoma epithelial cell-like) whole cell lysate 10ug
  Lane 2: B16-F0 (mouse melanoma epithelial cell-like) whole cell lysate
  Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab137078 in B16-F0 whole cell lysate

  Exposure time: 1 second

  Observed MW: 85-100 KDa

  All lanes: Immunoprecipitation - Anti-Melanoma gp100 antibody [EP4863(2)] (ab137078)

  Predicted band size: 70 kDa

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  Flow Cytometry (Intracellular) - Anti-Melanoma gp100 antibody [EP4863(2)] (ab137078)

  Overlay histogram showing B16-F0 (Mouse skin melanoma cell) cells stained with ab137078 (red line). The cells were fixed with 4% paraformaldehydeand then permeabilized with 90% methanol. The cells were then incubated with ab137078 at 1/40 dilution. The secondary antibody used wasGoat anti rabbit IgG (Alexa Fluor^® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) at 1/2000 dilution. Isotype control antibody (black line) was Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730). Unlabelled control (blue line) was cells without incubation with primary antibody and secondary antibody.
  

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  Immunocytochemistry/ Immunofluorescence - Anti-Melanoma gp100 antibody [EP4863(2)] (ab137078)

  ab137078 staining Melanoma gp100 in B16-F0 (mouse skin) cells by ICC (Immunocytochemistry). Cells were fixed with 4% Paraformaldehyde and permeabilized with 0.1% Triton X-100. Samples were incubated with primary antibody at a dilution of 1/50. A goat anti rabbit IgG (Alexa Fluor® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) was used as the secondary antibody at a dilution of 1/1000. Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 was used as a counterstain at a dilution of 1/200. DAPI was used as a nuclear counterstain.

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Melanoma gp100 antibody [EP4863(2)] (ab137078)

  ab137078 staining Melanoma gp100 in human melanoma tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde and antigen retrieval was by heat mediation in a EDTA buffer. Samples were incubated with primary antibody at a dilution of 1/250 for 30 minutes at room temperature. Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101) was used as the secondary antibody.
  

  The immunostaining staining was performed on a Leica Biosystems BOND® RX instrument.

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  Western blot - Anti-Melanoma gp100 antibody [EP4863(2)] (ab137078)

  Exposure time: 18-0 seconds
  

  Observed MW: 85-100 KDa

  The molecular weights observed are consistent with what have been described in PMID: 16492709 and PMID: 7961886

  All lanes: Western blot - Anti-Melanoma gp100 antibody [EP4863(2)] (ab137078) at 1/1000 dilution

  All lanes: Human melanoma lysate 15μg

  Secondary

  All lanes: Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

  Predicted band size: 70 kDa

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  Western blot - Anti-Melanoma gp100 antibody [EP4863(2)] (ab137078)

  Exposure time: 50 seconds
  

  Observed MW: 85-100 KDa

  The molecular weights observed are consistent with what have been described in PMID: 16492709 and PMID: 7961886

  All lanes: Western blot - Anti-Melanoma gp100 antibody [EP4863(2)] (ab137078) at 1/10000 dilution

  All lanes: B16-F0 (Mouse melanoma epithelial cell-like) whole cell lysate at 15 µg

  Secondary

  All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution

  Predicted band size: 70 kDa

  Exposure time: 50 seconds
  

  Observed MW: 85-100 KDa

  The molecular weights observed are consistent with what have been described in PMID: 16492709 and PMID: 7961886

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab137078).

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  Flow Cytometry (Intracellular) - Anti-Melanoma gp100 antibody [EP4863(2)] (ab137078)

  Flow cytometry overlay histogram showing left B16-F10 positive cells and right negative Raw264.7 stained with ab137078 (red line). The cells were fixed with 80% methanol (5 mins) and then permeabilised with 0.1% PBS-Triton X-100 for 15 mins. The cells were then incubated in 1x PBS containing 10μg/ml anti CD16/CD32 and 10% normal goat serum to block FC receptors and non-specific protein-protein interaction followed by the antibody (ab137078) (1x 10⁶ in 100μl at 1.0μg/ml (1/500 dilution)) for 30 mins at 22°C.

  The secondary antibody Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed was incubated at 1/4000 dilution for 30 mins at 22°C

  Isotype control antibody (black line) was Recombinant Rabbit IgG, monoclonal [EPR25A] - Isotype Control (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) used at the same concentration and conditions as the primary antibody. Unlabelled sample (blue line) was also used as a control.

  Acquisition of >5000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter.

  This antibody gave a positive signal in B16-F10 Fixed with 4% formaldehyde (10 min) / permeabilised with 0.1% PBS-Triton X-100 for 15 minss under the same conditions.

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  Flow Cytometry (Intracellular) - Anti-Melanoma gp100 antibody [EP4863(2)] (ab137078)

  Flow cytometry overlay histogram showing left Malme-3M positive cells and right negative A-375 stained with ab137078 (red line). The cells were fixed with 4% formaldehyde (10 min) and then permeabilised with 0.1% PBS-Triton X-100 for 15 mins. The cells were then incubated in 1x PBS containing 10% normal goat serum to block non-specific protein-protein interaction followed by the antibody (ab137078) (1x 10⁶ in 100μl at 1.0μg/ml (1/500 dilution)) for 30 mins at 22°C.

  The secondary antibody Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed was incubated at 1/4000 dilution for 30 mins at 22°C

  Isotype control antibody (black line) was Recombinant Rabbit IgG, monoclonal [EPR25A] - Isotype Control (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) used at the same concentration and conditions as the primary antibody. Unlabelled sample (blue line) was also used as a control.

  Acquisition of >5000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter.