Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-DIAPH1 antibody [EPR7948] - BSA and Azide free (ab248325)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR7948] to DIAPH1 - BSA and Azide free
  • Suitable for: ICC, IHC-P, WB, Flow Cyt
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-DIAPH1 antibody [EPR7948] - BSA and Azide free
    See all DIAPH1 primary antibodies

  • Description

    Rabbit monoclonal [EPR7948] to DIAPH1 - BSA and Azide free

  • Host species

    Rabbit

  • Tested applications

    Suitable for: ICC, IHC-P, WB, Flow Cytmore details

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

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

  • Positive control

    • WB: HCT116 and 293T cell lysates' HEK-293, RAW 264.7 and Human brain lysates, PC-12 and MCF7 lysates; Flow Cyt: HeLa cells; ICC: HeLa cells; IHC-P: Human, rat, and mouse kidney tissue sections.

  • General notes

    Ab248325 is the carrier-free version of ab129167. 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

    ab248325 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.

    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.

    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-DIAPH1 antibody [EPR7948] - BSA and Azide free (ab248325)
    Western blot - Anti-DIAPH1 antibody [EPR7948] - BSA and Azide free (ab248325)
    All lanes : Anti-DIAPH1 antibody [EPR7948] (ab129167) at 1/10000 dilution (Purified)

    Lane 1 : PC-12 (Rat adrenal gland pheochromocytoma ) whole cell lysate
    Lane 2 : MCF7 (Human breast adenocarcinoma 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: 141 kDa

  • Western blot - Anti-DIAPH1 antibody [EPR7948] - BSA and Azide free (ab248325)
    Western blot - Anti-DIAPH1 antibody [EPR7948] - BSA and Azide free (ab248325)
    All lanes : Anti-DIAPH1 antibody [EPR7948] (ab129167) at 1/1000 dilution (Purified)

    Lane 1 : HEK-293 (Human embryonic kidney epithelial cell) whole cell lysate
    Lane 2 : Human brain lysate
    Lane 3 : RAW 264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) 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: 141 kDa

  • Western blot - Anti-DIAPH1 antibody [EPR7948] - BSA and Azide free (ab248325)
    Western blot - Anti-DIAPH1 antibody [EPR7948] - BSA and Azide free (ab248325)
    All lanes : Anti-DIAPH1 antibody [EPR7948] (ab129167) at 1/1000 dilution

    Lane 1 : Wild-type HEK-293T cell lysate
    Lane 2 : DIAPH1 knockout HEK-293T cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 141 kDa
    Observed band size: 150 kDa
    why is the actual band size different from the predicted?



    This data was developed using the same antibody clone in a different buffer formulation (ab129167).

    Lanes 1- 2: Merged signal (red and green). Green - ab129167 observed at 150 kDa. Red - Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) observed at 50 kDa.

     ab129167 was shown to react with DIAPH1 in wild-type HEK-293T cells in western blot. Loss of signal was observed when knockout cell line ab266120 (knockout cell lysate ab257411) was used. Wild-type HEK-293T and DIAPH1 knockout HEK-293T 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. ab129167 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) overnight at 4°C at a 1 in 1000 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 - Anti-DIAPH1 antibody [EPR7948] - BSA and Azide free (ab248325)
    Immunocytochemistry - Anti-DIAPH1 antibody [EPR7948] - BSA and Azide free (ab248325)
    This data was developed using ab129167, the same antibody clone in a different buffer formulation.
    Immunocytochemistry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling DIAPH1 with Purified ab129167 at 1/50 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 dilution (2.5 µg/mL). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1/1000 dilution (2 µg/mL). DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DIAPH1 antibody [EPR7948] - BSA and Azide free (ab248325)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DIAPH1 antibody [EPR7948] - BSA and Azide free (ab248325)
    This data was developed using ab129167, the same antibody clone in a different buffer formulation.
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human kidney tissue sections labeling DIAPH1 with Purified ab129167 at 1/100 dilution (5.96 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-DIAPH1 antibody [EPR7948] - BSA and Azide free (ab248325)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DIAPH1 antibody [EPR7948] - BSA and Azide free (ab248325)
    This data was developed using ab129167, the same antibody clone in a different buffer formulation.
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse kidney tissue sections labeling DIAPH1 with Purified ab129167 at 1/100 dilution (5.96 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-DIAPH1 antibody [EPR7948] - BSA and Azide free (ab248325)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DIAPH1 antibody [EPR7948] - BSA and Azide free (ab248325)
    This data was developed using ab129167, the same antibody clone in a different buffer formulation.
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat kidney tissue sections labeling DIAPH1 with Purified ab129167 at 1/100 dilution (5.96 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.
  • Flow Cytometry - Anti-DIAPH1 antibody [EPR7948] - BSA and Azide free (ab248325)
    Flow Cytometry - Anti-DIAPH1 antibody [EPR7948] - BSA and Azide free (ab248325)

    This data was developed using ab129167, the same antibody clone in a different buffer formulation.

    Flow Cytometry analysis of HeLa (human epithelial cell line from cervix adenocarcinoma) cells labeling DIAPH1 with purified ab129167 at 1/50 dilution. Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) at 1/2000 was used as the secondary antibody (red). Rabbit monoclonal IgG (ab172730) was used as the isotype control (black). Cells without incubation with primary and secondary antibodies were used as the unlabeled control (blue).

  • Anti-DIAPH1 antibody [EPR7948] - BSA and Azide free (ab248325)
    Anti-DIAPH1 antibody [EPR7948] - BSA and Azide free (ab248325)

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