Anti-VPS35 antibody [EPR11501(B)] - BSA and Azide free

6 product images

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  Immunocytochemistry/ Immunofluorescence - Anti-VPS35 antibody [EPR11501(B)] - BSA and Azide free (ab240141)

  Anti-VPS35 antibody [EPR11501(B)] ab157220 staining VPS35 in wild-type HAP1 cells (top panel) and VPS35 knockout HAP1 cells (bottom panel). The cells were fixed with 4% formaldehyde (10min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with Anti-VPS35 antibody [EPR11501(B)] ab157220 at 1/250 dilution and Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 at 1/250 dilution (shown in pseudo colour red) overnight at +4°C, followed by a further incubation at room temperature for 1h with a goat secondary antibody to Rabbit IgG (Alexa Fluor® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 2 μg/ml (shown in green). Nuclear DNA was labelled in blue with DAPI.

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-VPS35 antibody [EPR11501(B)] ab157220).

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-VPS35 antibody [EPR11501(B)] - BSA and Azide free (ab240141)

  Immunohistochemical analysis of paraffin-embedded Human kidney tissue labeling VPS35 with Anti-VPS35 antibody [EPR11501(B)] ab157220 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-VPS35 antibody [EPR11501(B)] ab157220).

  Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-VPS35 antibody [EPR11501(B)] - BSA and Azide free (ab240141)

  Immunohistochemical analysis of paraffin-embedded Human ovarian carcinoma tissue labeling VPS35 with Anti-VPS35 antibody [EPR11501(B)] ab157220 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-VPS35 antibody [EPR11501(B)] ab157220).

  Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

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  Flow Cytometry (Intracellular) - Anti-VPS35 antibody [EPR11501(B)] - BSA and Azide free (ab240141)

  Intracellular flow cytometric analysis of permeabilized MOLT4 cells labeling VPS35 with Anti-VPS35 antibody [EPR11501(B)] ab157220 at 1/10 dilution (red) or a rabbit IgG (negative) (green). This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-VPS35 antibody [EPR11501(B)] ab157220).
  

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-VPS35 antibody [EPR11501(B)] - BSA and Azide free (ab240141)

  This data was developed using Anti-PMS2 antibody [EPR3947] ab110638, the same antibody clone in a different buffer formulation.

  Immunohistochemical analysis of formalin fixed paraffin embedded human kidney labelling VPS35 with Anti-VPS35 antibody [EPR11501(B)] ab157220 at a concentration of 0.01µg/ml. The immunostaining was performed on a Ventana DISCOVERY ULTRA (Roche Tissue Diagnostics) instrument with a OptiView DAB IHC Detection Kit. Heat mediated antigen retrieval was performed with DISCOVERY cell conditioning solution (CC1) 100°C, pH8.5 for 32mins.

  Anti-VPS35 antibody [EPR11501(B)] ab157220 Anti-VPS35 antibody [EPR11501(B)] was incubated for 16mins at 37°C. Sections were counterstained with Hematoxylin II. Image inset shows absence of staining in secondary antibody only control.

  Customers are encouraged to optimise antigen retrieval conditions, antibody concentration, incubation times and temperature for best results in their own IHC assay workflow (automated and manual).

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  Western blot - Anti-VPS35 antibody [EPR11501(B)] - BSA and Azide free (ab240141)

  This data was developed using Anti-VPS35 antibody [EPR11501(B)] ab157220, the same antibody clone in a different buffer formulation.

  Anti-VPS35 antibody [EPR11501(B)] ab157220 was shown to react with VPS35 in wild-type HAP1 cells in Western blot with loss of signal observed in a VPS35 knockout cell line. Wild-type HAP1 and VPS35 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 5% milk in TBST for 1 hr before incubation with Anti-VPS35 antibody [EPR11501(B)] ab157220 overnight at 4 °C at a 1/1000 dilution. Blots were incubated with secondary antibodies at 0.2 µg/mL before imaging.
  
  These data were provided by YCharOS Inc., an open science company with the mission of characterizing commercially available antibody reagents for all human proteins. Abcam and YCharOS are working together to help address the reproducibility crisis by enabling the life science community to better evaluate commercially available antibodies.

  All lanes: Western blot - Anti-VPS35 antibody [EPR11501(B)] (Anti-VPS35 antibody [EPR11501(B)] ab157220) at 1/1000 dilution

  Lane 1: Wild-type HAP1 lysate at 20 µg

  Lane 2: VPS35 knock-out HAP1 lysate at 20 µg