Anti-RAB7 antibody [EPR7589] - BSA and Azide free

6 product images

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  Western blot - Anti-RAB7 antibody [EPR7589] - BSA and Azide free (ab214806)

  Lanes 1 - 2: Merged signal (red and green). Green - Anti-RAB7 antibody [EPR7589] ab137029 observed at 23 kDa. Red - loading control Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55 kDa.
  

  Anti-RAB7 antibody [EPR7589] ab137029 was shown to react with RAB7 in wild-type HeLa cells in Western blot with loss of signal observed in RAB7A knockout cell line Human RAB7A (RAB7) knockout HeLa cell line ab255423 (RAB7A knockout cell lysate Human RAB7A (RAB7) knockout HeLa cell lysate ab263831). Wild-type HeLa and RAB7A knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween^®) before incubation with Anti-RAB7 antibody [EPR7589] ab137029 and Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) overnight at 4 °C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated 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 h at room temperature before imaging.

  All lanes: Western blot - Anti-RAB7 antibody [EPR7589] - BSA and Azide free (ab214806)

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

  Lane 2: RAB7A knockout HeLa cell lysate at 20 µg

  Performed under reducing conditions.

  Observed band size: 23 kDa

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  Immunocytochemistry/ Immunofluorescence - Anti-RAB7 antibody [EPR7589] - BSA and Azide free (ab214806)

  Immunocytochemistry/Immunofluorescence analysis of HeLa (human cervix adenocarcinoma) labelling RAB7 with purified Anti-RAB7 antibody [EPR7589] ab137029 at 1/500. Cells were fixed with 100% methanol. An Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077). Nuclei counterstained with DAPI (blue).

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-RAB7 antibody [EPR7589] ab137029).

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  Flow Cytometry (Intracellular) - Anti-RAB7 antibody [EPR7589] - BSA and Azide free (ab214806)

  Overlay histogram showing HAP1 wildtype (green line) and HAP1-RAB7 knockout cells (red line) stained with Anti-RAB7 antibody [EPR7589] ab137029. The cells were fixed with 4% formaldehyde (10 min) and then permeabilized with 0.1% PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS / 10% normal goat serum to block non-specific protein-protein interactions followed by the antibody (Anti-RAB7 antibody [EPR7589] ab137029, 0.1μg/ml) for 30 min at 22°C. The secondary antibody used was Alexa Fluor^® 488 goat anti-rabbit IgG (H&L) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 1/2000 dilution for 30 min at 22°C. A rabbit IgG1 isotype control antibody (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) was used at the same concentration and conditions as the primary antibody (HAP1 wildtype - black line, HAP1-RAB7 knockout - grey line). Unlabelled sample was also used as a control (this line is not shown for the purpose of simplicity). Acquisition of >5,000 events were collected using a 50 mW Blue laser (488nm) and 530/30 bandpass filter. This antibody can also be used in HAP1 cells fixed with 80% methanol (5 min) permeabilized with 0.1% PBS-Triton X-100 for 15 min under the same conditions.
  

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-RAB7 antibody [EPR7589] ab137029).

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RAB7 antibody [EPR7589] - BSA and Azide free (ab214806)

  Immunohistochemical analysis of paraffin embedded Human kidney tissue labelling RAB7 with Anti-RAB7 antibody [EPR7589] ab137029 at 1/50 dilution.
  

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-RAB7 antibody [EPR7589] ab137029).

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

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  This image is courtesy of a customer review submitted by Alina Macovei

  Immunocytochemistry/ Immunofluorescence - Anti-RAB7 antibody [EPR7589] - BSA and Azide free (ab214806)

  Anti-RAB7 antibody [EPR7589] ab137029 staining RAB7 in Human HepaRG cell by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with formaldehyde, permeabilized with 0.1% Triton X-100 in PBS and blocked with 1% milk for 30 minutes at room temperature. Samples were incubated with primary antibody (1/200 in 1% milk) for 30 minutes. An Alexa Fluor®488-conjugated Donkey anti-rabbit IgG polyclonal (1/400) was used as the secondary antibody.

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-RAB7 antibody [EPR7589] ab137029).

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  Flow Cytometry (Intracellular) - Anti-RAB7 antibody [EPR7589] - BSA and Azide free (ab214806)

  Intracellular Flow Cytometry analysis of A431 (human epidermoid carcinoma) cells labeling RAB7 with purified Anti-RAB7 antibody [EPR7589] ab137029 at 1/20 dilution (10ug/ml) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor^® 488) (1/2000 dilution) was used as the secondary antibody. Rabbit monoclonal IgG (Black) was used as the isotype control, cells without incubation with primary antibody and secondary antibody (Blue) was used as the unlabeled control.
  

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-RAB7 antibody [EPR7589] ab137029).