Anti-PLCG 2 antibody [EPR5914-34]

7 product images

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PLCG 2 antibody [EPR5914-34] (ab133522)

  Immunohistochemistry analysis of Paraffin Embedded Human gastric adenocarcinoma tissue labelling PLCG2 with ab133522 at 1/100 dilution.
  

  Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

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  Western blot - Anti-PLCG 2 antibody [EPR5914-34] (ab133522)

  All lanes: Western blot - Anti-PLCG 2 antibody [EPR5914-34] (ab133522) at 1/10000 dilution

  Lane 1: Raji cell lysates at 10 µg

  Lane 2: Ramos cell lysates at 10 µg

  Secondary

  All lanes: HRP-labelled Goat anti-Rabbit at 1/2000 dilution

  Predicted band size: 148 kDa

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  Flow Cytometry (Intracellular) - Anti-PLCG 2 antibody [EPR5914-34] (ab133522)

  Flow Cytometry analysis of Human Peripheral Blood Mononuclear Cell (PBMC) cells labeling PLCG 2 with purified ab133522 at 1/1000 dilution (1 μg/mL) (Red). Cells were fixed with 2% Paraformaldehyde and permeabilised with 0.1% Tween-20. A Goat anti rabbit IgG (Alexa Fluor^® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Left).
  Cells were surface stained with anti-CD19 conjugated to PE/Cy7, then fixed with 2% PFA followed by intracellularly stained with rabbit IgG or ab133522. Gated on lymphocytes.

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  OI-RD Scanning - Anti-PLCG 2 antibody [EPR5914-34] (ab133522)

  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.

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  Immunoprecipitation - Anti-PLCG 2 antibody [EPR5914-34] (ab133522)

  This data was developed using Anti-PLCG 2 antibody [EPR5914-34] - BSA and Azide free ab248535, the same antibody clone in a different buffer formulation.
  Western blot: antibody description (Anti-PLCG 2 antibody [EPR5914-34] - BSA and Azide free ab248535) staining at 10,000 dilution, shown in Black. In Western blot, Anti-PLCG 2 antibody [EPR5914-34] - BSA and Azide free ab248535 was shown to bind specifically to PLCG2. An enriched band was observed at 160 kDa in samples pulled down with Anti-PLCG 2 antibody [EPR5914-34] - BSA and Azide free ab248535 with no band observed in Isotype control sample. Following IP, 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 before development with a high-sensitivity ECL substrate kit and imaged. Secondary antibodies used was VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) at 1/1000 dilution.

  Lanes 1 - 2: Immunoprecipitation - Anti-PLCG 2 antibody [EPR5914-34] - BSA and Azide free (Anti-PLCG 2 antibody [EPR5914-34] - BSA and Azide free ab248535) at 1/10000 dilution

  Lanes 1 - 2: Immunoprecipitation - Anti-PLCG 2 antibody [EPR5914-34] (ab133522)

  Lane 1: HAP1 Input cell lysate at 10 µg

  Lane 2: HAP1 pulldown with Anti-PLCG 2 antibody [EPR5914-34] - BSA and Azide free ab248535 at 10 µg

  Lane 3: HAP1 pulldown with Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730 Isotype control at 10 µg

  Secondary

  All lanes: Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) at 1/1000 dilution

  Developed using the ECL technique.

  Observed band size: 160 kDa

  Exposure time: 4min

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  Western blot - Anti-PLCG 2 antibody [EPR5914-34] (ab133522)

  Western blot: Anti-PLCG2 antibody [EPR5914-34] (ab133522) staining at 1/10000 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, ab133522 was shown to bind specifically to PLCG2. A band was observed at 150 kDa in wild-type HCT 116 cell lysates with no signal observed at this size in PLCG2 knockout cell line. To generate this image, wild-type and PLCG2 knockout 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 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

  All lanes: Western blot - Anti-PLCG 2 antibody [EPR5914-34] (ab133522) at 1/10000 dilution

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

  Lane 2: Western blot - Human PLCG2 knockout HCT116 cell line (Human PLCG2 knockout HCT116 cell line ab301166)

  Lane 2: PLCG2 knockout HCT 116 cell lysate at 20 µg

  Secondary

  Lanes 1 - 2: Goat anti-Rabbit IgG H&L 800CW at 1/20000 dilution

  Lanes 1 - 2: Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

  Performed under reducing conditions.

  Observed band size: 150 kDa

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  Western blot - Anti-PLCG 2 antibody [EPR5914-34] (ab133522)

  Anti-PLCG2 antibody [EPR5914-34] (ab133522) staining at 1/10000 dilution, shown in green; Mouse anti-CANX [CANX/1543] (Anti-Calnexin antibody [CANX/1543] ab238078) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab133522 was shown to bind specifically to PLCG2. A band was observed at 148 kDa in wild-type HAP1 cell lysates with no signal observed at this size in PLCG2 knockout cell line. To generate this image, wild-type and PLCG2 knockout HAP1 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 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

  All lanes: Western blot - Anti-PLCG 2 antibody [EPR5914-34] (ab133522) at 1/10000 dilution

  All lanes: Western blot at 20 µg

  Secondary

  All lanes: Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

  Performed under reducing conditions.

  Predicted band size: 148 kDa

  Observed band size: 148 kDa