Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-eIF4EBP1 antibody [Y329] - BSA and Azide free (ab173370)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [Y329] to eIF4EBP1 - BSA and Azide free
  • Suitable for: Flow Cyt, IHC-P, IP, WB, ICC
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-eIF4EBP1 antibody [Y329] - BSA and Azide free
    See all eIF4EBP1 primary antibodies

  • Description

    Rabbit monoclonal [Y329] to eIF4EBP1 - BSA and Azide free

  • Host species

    Rabbit

  • Tested Applications & Species

    Application Species
    Flow Cyt
    Human
    ICC
    Human
    IHC-P
    Rat
    IP
    Human
    WB
    Human
    See all applications and species data

  • Immunogen

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

  • Positive control

    • WB: HeLa, HAP1, K562 and 293 cell lysates. K-562 and HEK-293 whole cell lysates. Mouse and rat skeletal muscle lysate. Rat L6 whole cell lysate. IHC-P: Human colon cancer tissue. Rat and mouse spleen tissues. Flow Cyt: HAP1-WT, HEK-293, and HT1080 cells. IP: HEK-293 whole cell lysate. ICC: HeLa cells.

  • General notes

    Ab173370 is the carrier-free version of ab32024. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

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

    ab173370 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    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.

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.

    One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.

    Learn more here.

Properties

Images

  • Western blot - Anti-eIF4EBP1 antibody [Y329] - BSA and Azide free (ab173370)
    Western blot - Anti-eIF4EBP1 antibody [Y329] - BSA and Azide free (ab173370)
    All lanes : Anti-eIF4EBP1 antibody [Y329] (ab32024) at 1/2000 dilution (Purified)

    Lane 1 : HEK-293 (Human embryonic kidney epithelial cell) whole cell lysate
    Lane 2 : K-562 (Human chronic myelogenous leukemia lymphoblast) whole cell lysate
    Lane 3 : Mouse skeletal muscle lysate
    Lane 4 : Rat skeletal muscle lysate
    Lane 5 : L6 (Rat skeletal muscle myoblast) 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: 13 kDa



    This data was developed using ab32024, the same antibody clone in a different buffer formulation.

  • Immunoprecipitation - Anti-eIF4EBP1 antibody [Y329] - BSA and Azide free (ab173370)
    Immunoprecipitation - Anti-eIF4EBP1 antibody [Y329] - BSA and Azide free (ab173370)

    This data was developed using ab32024, the same antibody clone in a different buffer formulation.
    Purified ab32024 at 1/20 dilution (2µg) immunoprecipitating eIF4EBP1 in HEK-293 whole cell lysate.
    Lane 1 (input): HEK-293 (Human embryonic kidney epithelial cell) whole cell lysate 10µg
    Lane 2 (+): ab32024 + HEK-293 whole cell lysate.
    Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab32024 in HEK-293 whole cell lysate.
    VeriBlot for IP Detection Reagent (HRP) (ab131366) (1/1000 dilution) was used for Western blotting.
    Blocking Buffer and concentration: 5% NFDM/TBST.
    Diluting buffer and concentration: 5% NFDM/TBST.
    Observed band size: 15-20 kDa

  • Flow Cytometry - Anti-eIF4EBP1 antibody [Y329] - BSA and Azide free (ab173370)
    Flow Cytometry - Anti-eIF4EBP1 antibody [Y329] - BSA and Azide free (ab173370)
    This data was developed using ab32024, the same antibody clone in a different buffer formulation.
    Flow Cytometry analysis of HEK-293 (Human embryonic kidney epithelial cell) cells labeling eIF4EBP1 with purified ab32024 at 1/20 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).
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-eIF4EBP1 antibody [Y329] - BSA and Azide free (ab173370)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-eIF4EBP1 antibody [Y329] - BSA and Azide free (ab173370)
    This data was developed using ab32024, the same antibody clone in a different buffer formulation.
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat spleen tissue sections labeling eIF4EBP1 with purified ab32024 at 1/200 dilution (0.53 µg/mL). Heat mediated antigen retrieval 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.
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-eIF4EBP1 antibody [Y329] - BSA and Azide free (ab173370)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-eIF4EBP1 antibody [Y329] - BSA and Azide free (ab173370)
    This data was developed using ab32024, the same antibody clone in a different buffer formulation.
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse spleen tissue sections labeling eIF4EBP1 with purified ab32024 at 1/200 dilution (0.53 µg/mL). Heat mediated antigen retrieval 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.
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-eIF4EBP1 antibody [Y329] - BSA and Azide free (ab173370)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-eIF4EBP1 antibody [Y329] - BSA and Azide free (ab173370)
    This data was developed using ab32024, the same antibody clone in a different buffer formulation.
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human colon cancer tissue sections labeling eIF4EBP1 with purified ab32024 at 1/200 dilution (0.53 µg/mL). Heat mediated antigen retrieval 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.
  • Western blot - Anti-eIF4EBP1 antibody [Y329] - BSA and Azide free (ab173370)
    Western blot - Anti-eIF4EBP1 antibody [Y329] - BSA and Azide free (ab173370)
    All lanes : Anti-eIF4EBP1 antibody [Y329] (ab32024) at 1/5000 dilution

    Lane 1 : Wild-type HeLa cell lysate
    Lane 2 : EIF4EBP1 knockout HeLa cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 13 kDa
    Observed band size: 13 kDa



    This data was developed using the same antibody clone in a different buffer formulation (ab32024).

    Lanes 1-2: Merged signal (red and green). Green - ab32024 observed at 13 kDa. Red - loading control ab8245 observed at 37 kDa.

     ab32024 Anti-eIF4EBP1 antibody [Y329] was shown to specifically react with eIF4EBP1 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab264784 (knockout cell lysate ab257146) was used. Wild-type and eIF4EBP1 knockout samples were subjected to SDS-PAGE. ab32024 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 5000 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.

     

     

  • Flow Cytometry - Anti-eIF4EBP1 antibody [Y329] - BSA and Azide free (ab173370)
    Flow Cytometry - Anti-eIF4EBP1 antibody [Y329] - BSA and Azide free (ab173370)

    Overlay histogram showing HAP1 wildtype (green line) and HAP1-EIF4EBP1   knockout cells (red line) stained with ab32024. The cells were fixed with 80% methanol (5 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 (ab32024, 0.1µg/ml) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) presorbed (ab150081) at 1/2000 dilution for 30 min at 22°C.

    A rabbit IgG isotype control antibody  (ab172730) was used at the same concentration and conditions as the primary antibody (HAP1 wildtype - black line, HAP1-EIF4EBP1  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 4% formaldehyde (10 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 (ab32024).

  • Immunocytochemistry - Anti-eIF4EBP1 antibody [Y329] - BSA and Azide free (ab173370)
    Immunocytochemistry - Anti-eIF4EBP1 antibody [Y329] - BSA and Azide free (ab173370)

    Immunocytochemistry/Immunofluorescence analysis of HeLa (Human epithelial cell line from cervix adenocarcinoma) labeling eIF4EBP1 with Purified ab32024 at 1/500 dilution (5 µg/ml). Cells were fixed with 4% PFA and permeabilized with 0.1% tritonX-100. ab150077 Goat anti rabbit IgG(Alexa Fluor® 488) at 1/1000 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI. PBS was used instead of the primary antibody as the negative control.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32024).

  • Anti-eIF4EBP1 antibody [Y329] - BSA and Azide free (ab173370)
    Anti-eIF4EBP1 antibody [Y329] - BSA and Azide free (ab173370)

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