Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-Raf1 antibody [EP4969] - BSA and Azide free (ab236003)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EP4969] to Raf1 - BSA and Azide free
  • Suitable for: Flow Cyt (Intra), ICC/IF, WB
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-Raf1 antibody [EP4969] - BSA and Azide free
    See all Raf1 primary antibodies

  • Description

    Rabbit monoclonal [EP4969] to Raf1 - BSA and Azide free

  • Host species

    Rabbit

  • Tested applications

    Suitable for: Flow Cyt (Intra), ICC/IF, WBmore details

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

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

  • Positive control

    • WB: HAP1, HeLa, K-562, RAW 264.7, C2C12, NIH/3T3, and PC-12 cell lysates. ICC/IF: HeLa and K562 cells. Flow Cyt (intra): K562 cells.

  • General notes

    ab236003 is the carrier-free version of ab181115.

    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-Raf1 antibody [EP4969] - BSA and Azide free (ab236003)
    Western blot - Anti-Raf1 antibody [EP4969] - BSA and Azide free (ab236003)
    All lanes : Anti-Raf1 antibody [EP4969] - N-terminal (ab181115) at 1/20000 dilution

    Lane 1 : Wild-type HeLa cell lysate
    Lane 2 : RAF1 knockout HeLa cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 73 kDa
    Observed band size: 73 kDa



    This data was developed using the same antibody clone in a different buffer formulation (ab181115).

      Lanes 1- 2: Merged signal (red and green). Green - ab181115 observed at 73 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

     ab181115 was shown to react with Raf1 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab264978 (knockout cell lysate ab257126) was used. Wild-type HeLa and RAF1 knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab181115 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 20000 Dilution and a 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.

  • Immunocytochemistry/ Immunofluorescence - Anti-Raf1 antibody [EP4969] - BSA and Azide free (ab236003)
    Immunocytochemistry/ Immunofluorescence - Anti-Raf1 antibody [EP4969] - BSA and Azide free (ab236003)
    Immunocytochemistry/ Immunofluorescence analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling Raf1 with purified ab181115 at 1/100 dilution (10 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 (ab181115)
  • Flow Cytometry - Anti-Raf1 antibody [EP4969] - BSA and Azide free (ab236003)
    Flow Cytometry - Anti-Raf1 antibody [EP4969] - BSA and Azide free (ab236003)

    Flow cytometry analysis of K-562 (Human chronic myelogenous leukemia lymphoblast) cells labelling with ab181115 at 1/100 dilution (10.79 µg/mL) (red). Cells were fixed with 4% paraformaldehyde . Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1/2000 dilution. Isotype control - 90% methanol . Unlabeled control - Rabbit monoclonal IgG (ab172730) / Black.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab181115).

  • Western blot - Anti-Raf1 antibody [EP4969] - BSA and Azide free (ab236003)
    Western blot - Anti-Raf1 antibody [EP4969] - BSA and Azide free (ab236003)
    Lanes 1-2 : Anti-Raf1 antibody [EP4969] - N-terminal (ab181115) at 1/1000 dilution
    Lanes 3-4 : Anti-beta Actin antibody [mAbcam 8226] - Loading Control (ab8226) at 1/1000 dilution


    Lanes 1 & 3 & 5 : Wild-type HAP1 cell lysate
    Lanes 2 & 4 & 6 : Raf1 knockout HAP1 cell lysate

    Lysates/proteins at 20 µg per lane.

    Predicted band size: 73 kDa



    Lanes 1 and 2: Green signal from target – ab181115 observed at 74 kDa
    Lanes 3 and 4: Red signal from loading control – ab8226 observed at 42 kDa
    Lanes 5 and 6: Merged (red and green) signal

    ab181115 was shown to specifically react with Raf1 when Raf1 knockout samples were used. Wild-type and Raf1 knockout samples were subjected to SDS-PAGE. ab181115 and ab8226 (loading control to beta actin) were both diluted 1/1000 and incubated overnight at 4ºC. 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 1/10000 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 (ab181115).

  • Immunocytochemistry/ Immunofluorescence - Anti-Raf1 antibody [EP4969] - BSA and Azide free (ab236003)
    Immunocytochemistry/ Immunofluorescence - Anti-Raf1 antibody [EP4969] - BSA and Azide free (ab236003)

    Immunofluorescent analysis of acetone-fixed K562 cells labeling Raf1 with ab181115 at 1/250 dilution, followed by Goat anti rabbit IgG (Alexa Fluor® 488) secondary antibody at 1/200 dilution. Counter stained with Dapi.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab181115).

  • Anti-Raf1 antibody [EP4969] - BSA and Azide free (ab236003)
    Anti-Raf1 antibody [EP4969] - BSA and Azide free (ab236003)

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