Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-ULK3 antibody [EPR4888] - BSA and Azide free (ab232372)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR4888] to ULK3 - BSA and Azide free
  • Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IF
  • Knockout validated
  • Reacts with: Human

Overview

  • Product name

    Anti-ULK3 antibody [EPR4888] - BSA and Azide free
    See all ULK3 primary antibodies

  • Description

    Rabbit monoclonal [EPR4888] to ULK3 - BSA and Azide free

  • Host species

    Rabbit

  • Tested applications

    Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IFmore details

  • Species reactivity

    Reacts with: Human
    Does not react with: Mouse, Rat

  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • IHC-P: Human bladder carcinoma and human colon tissue; WB: Wild-type HAP1 whole cell lysate; HEK-293 and LNCaP whole cell lysates. Human heart and testis tissue lysates. ICC/IF: LNCaP cells; Flow Cyt (intra): LNCaP cells.

  • General notes

    ab232372 is the carrier-free version of ab124947.

    Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.

    This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

    Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with

    This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

Properties

Images

  • Western blot - Anti-ULK3 antibody [EPR4888] - BSA and Azide free (ab232372)
    Western blot - Anti-ULK3 antibody [EPR4888] - BSA and Azide free (ab232372)
    All lanes : Anti-ULK3 antibody [EPR4888] (ab124947) at 1/1000 dilution

    Lane 1 : Wild-type HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate
    Lane 2 : ULK3 knockout HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate
    Lane 3 : HAP1 whole cell lyate
    Lane 4 : LNCaP (Human prostate cancer cell line) whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution

    Predicted band size: 53 kDa
    Observed band size: 55 kDa
    why is the actual band size different from the predicted?



    This data was developed using ab124947, the same antibody clone in a different buffer formulation.

    Lanes 1-4: Merged signal (red and green). Green - ab124947 observed at 55 kDa. Red - loading control ab8245 observed at 36 kDa.

     ab124947 Anti-ULK3 antibody [EPR4888] was shown to specifically react with ULK3 in wild-type HEK-293T cells. Loss of signal was observed when knockout cell line ab266152 (knockout cell lysate ab258271) was used. Wild-type and ULK3 knockout samples were subjected to SDS-PAGE. ab124947 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ULK3 antibody [EPR4888] - BSA and Azide free (ab232372)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ULK3 antibody [EPR4888] - BSA and Azide free (ab232372)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human colon tissue sections labeling ULK3 with purified ab124947 at 1/50 dilution (7.84 µg/mL). Heat mediated antigen retrieval was performed using Bond™ Epitope Retrieval Solution 2 (pH 9.0). Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
    The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab124947).

  • Immunocytochemistry/ Immunofluorescence - Anti-ULK3 antibody [EPR4888] - BSA and Azide free (ab232372)
    Immunocytochemistry/ Immunofluorescence - Anti-ULK3 antibody [EPR4888] - BSA and Azide free (ab232372)

    Immunocytochemistry/ Immunofluorescence analysis of LNCaP (Human prostate carcinoma epithelial cell) cells labeling ULK3 with Purified ab124947 at 1:50 dilution (7.8 µg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1:200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1:1000 (2 µg/ml) dilution. DAPI (blue) was used as 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 (ab124947).

  • Western blot - Anti-ULK3 antibody [EPR4888] - BSA and Azide free (ab232372)
    Western blot - Anti-ULK3 antibody [EPR4888] - BSA and Azide free (ab232372)

    Lane 1: Wild-type HAP1 whole cell lysate (20 µg)
    Lane 2: ULK3 knockout HAP1 whole cell lysate (20 µg)
    Lane 3: Hek293 whole cell lysate (20 µg)
    Lanes 1 - 3: Merged signal (red and green). Green - ab124947 observed at 55 kDa. Red - loading control, ab8245, observed at 37 kDa.

    ab124947 (unpurified) was shown to specifically react with ULK3 in wild-type HAP1 cells along with additional cross-reactive bands. No band was observed when ULK3 knockout samples were examined. Wild-type and ULK3 knockout samples were subjected to SDS-PAGE. Ab124947 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/10,000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) ab216776 secondary antibodies at 1/10,000 dilution for 1 hour at room temperature before imaging.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab124947).

  • Flow Cytometry - Anti-ULK3 antibody [EPR4888] - BSA and Azide free (ab232372)
    Flow Cytometry - Anti-ULK3 antibody [EPR4888] - BSA and Azide free (ab232372)

    Flow Cytometry analysis of LNCaP (Human prostate carcinoma epithelial cell) cells labeling ULK3 with Purified ab124947 at 1:40 dilution (10 µg/ml) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) secondary antibody was used at 1:2000. 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 (ab124947).

  • Western blot - Anti-ULK3 antibody [EPR4888] - BSA and Azide free (ab232372)
    Western blot - Anti-ULK3 antibody [EPR4888] - BSA and Azide free (ab232372)
    All lanes : Anti-ULK3 antibody [EPR4888] (ab124947) at 1/1000 dilution (Purified)

    Lane 1 : Human heart lysate
    Lane 2 : Human testis lysate
    Lane 3 : LNCaP (Human prostate carcinoma epithelial cell) whole cell lysate
    Lane 4 : HEK-293 (Human embryonic kidney epithelial cell) whole cell lysate

    Lysates/proteins at 20 µg per lane.

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

    Predicted band size: 53 kDa
    Observed band size: 53 kDa



    Blocking/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 (ab124947).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ULK3 antibody [EPR4888] - BSA and Azide free (ab232372)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ULK3 antibody [EPR4888] - BSA and Azide free (ab232372)

    ab124947 (unpurified), at a 1/50 dilution, staining ULK3 in paraffin embedded Human bladder carcinoma tissue by Immunohistochemistry. Heat mediated antigen retrieval was performed with citrate buffer pH 6 before commencing with IHC staining protocol.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab124947).

  • Anti-ULK3 antibody [EPR4888] - BSA and Azide free (ab232372)
    Anti-ULK3 antibody [EPR4888] - BSA and Azide free (ab232372)

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