Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-SMARCD1 antibody [EPR23170-71] - BSA and Azide free (ab269463)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR23170-71] to SMARCD1 - BSA and Azide free
  • Suitable for: Flow Cyt (Intra), WB, ICC/IF, IP
  • Knockout validated
  • Reacts with: Mouse, Human

Overview

  • Product name

    Anti-SMARCD1 antibody [EPR23170-71] - BSA and Azide free
    See all SMARCD1 primary antibodies

  • Description

    Rabbit monoclonal [EPR23170-71] to SMARCD1 - BSA and Azide free

  • Host species

    Rabbit

  • Tested applications

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

  • Species reactivity

    Reacts with: Mouse, Human

  • Immunogen

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

  • Positive control

    • WB: HeLa, 293T, NIH/3T3 and Neuro-2a lysates. ICC/IF: HeLa and Neuro-2a cells. Flow Cyt (intra): HeLa and NIH/3T3 cells. IP: HeLa and NIH/3T3 cells.

  • General notes

    ab269463 is the carrier-free version of ab245222.

    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.

Images

  • Western blot - Anti-SMARCD1 antibody [EPR23170-71] - BSA and Azide free (ab269463)
    Western blot - Anti-SMARCD1 antibody [EPR23170-71] - BSA and Azide free (ab269463)
    All lanes : Anti-SMARCD1 antibody [EPR23170-71] (ab245222) 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 : SMARCD1 knockout HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate
    Lane 3 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
    Lane 4 : NIH/3T3 (Mouse embryonic fibroblast 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: 58 kDa
    Observed band size: 58 kDa



    This data was developed using ab245222, the same antibody clone in a different buffer formulation.

    Lanes 1-4: Merged signal (red and green). Green - ab245222 observed at 58 kDa. Red - loading control ab8245 observed at 36 kDa.

     ab245222 Anti-SMARCD1 antibody [EPR23170-71] was shown to specifically react with SMARCD1 in wild-type HEK-293T cells. Loss of signal was observed when knockout cell line ab266458 (knockout cell lysate ab259143) was used. Wild-type and SMARCD1 knockout samples were subjected to SDS-PAGE. ab245222 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.

  • Immunocytochemistry/ Immunofluorescence - Anti-SMARCD1 antibody [EPR23170-71] - BSA and Azide free (ab269463)
    Immunocytochemistry/ Immunofluorescence - Anti-SMARCD1 antibody [EPR23170-71] - BSA and Azide free (ab269463)

    Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized Neuro-2a (mouse neuroblastoma neuroblast) cells labelling SMARCD1 with ab245222 at 1/50 dilution, followed by ab150077 AlexaFluor®488 Goat anti-Rabbit secondary antibody at 1/1000 dilution (Green). Confocal image showing nuclear staining in Neuro-2a cells. ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).

    Secondary antibody only control: Secondary antibody is ab150077 AlexaFluor®488 Goat anti-Rabbit secondary at 1/1000 dilution.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab245222).

  • Immunoprecipitation - Anti-SMARCD1 antibody [EPR23170-71] - BSA and Azide free (ab269463)
    Immunoprecipitation - Anti-SMARCD1 antibody [EPR23170-71] - BSA and Azide free (ab269463)

    SMARCD1 was immunoprecipitated from 0.35 mg of NIH/3T3 (Mouse embryonic fibroblast) whole cell lysate with ab245222 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab245222 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used as secondary antibody at 1/5000 dilution.

    Lane 1: NIH/3T3 whole cell lysate 10 (Input). 
    Lane 2: ab245222 IP in NIH/3T3 whole cell lysate. 
    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab245222 in NIH/3T3 whole cell lysate.

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.
    Exposure time: 3 minutes.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab245222).

  • Flow Cytometry - Anti-SMARCD1 antibody [EPR23170-71] - BSA and Azide free (ab269463)
    Flow Cytometry - Anti-SMARCD1 antibody [EPR23170-71] - BSA and Azide free (ab269463)

    Flow cytometric analysis of 4% paraformaldehyde fixed, 90% methanol permeabilized HeLa (Human cervix adenocarcinoma epithelial cell) cells labelling SMARCD1 with ab245222 at 1/500 dilution (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab245222).

  • Immunoprecipitation - Anti-SMARCD1 antibody [EPR23170-71] - BSA and Azide free (ab269463)
    Immunoprecipitation - Anti-SMARCD1 antibody [EPR23170-71] - BSA and Azide free (ab269463)

    SMARCD1 was immunoprecipitated from 0.35 mg of HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate whole cell lysate with ab245222 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab245222 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used as secondary antibody at 1/5000 dilution.

    Lane 1: HeLa whole cell lysate 10 (Input). 
    Lane 2: ab245222 IP in HeLa whole cell lysate. 
    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab245222 in HeLa whole cell lysate.

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.
    Exposure time: 3 minutes.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab245222).

  • Flow Cytometry - Anti-SMARCD1 antibody [EPR23170-71] - BSA and Azide free (ab269463)
    Flow Cytometry - Anti-SMARCD1 antibody [EPR23170-71] - BSA and Azide free (ab269463)

    Flow cytometric analysis of 4% paraformaldehyde fixed, 90% methanol permeabilized NIH/3T3 (Mouse embryonic fibroblast) cells labelling SMARCD1 with ab245222 at 1/500 dilution (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab245222).

  • Immunocytochemistry/ Immunofluorescence - Anti-SMARCD1 antibody [EPR23170-71] - BSA and Azide free (ab269463)
    Immunocytochemistry/ Immunofluorescence - Anti-SMARCD1 antibody [EPR23170-71] - BSA and Azide free (ab269463)

    Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HeLa (human cervix adenocarcinoma epithelial cell) cells labelling SMARCD1 with ab245222 at 1/50 dilution, followed by ab150077 AlexaFluor®488 Goat anti-Rabbit secondary antibody at 1/1000 dilution (Green). Confocal image showing nuclear staining in HeLa cells. ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).

    Secondary antibody only control: Secondary antibody is ab150077 AlexaFluor®488 Goat anti-Rabbit secondary at 1/1000 dilution.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab245222).

  • Anti-SMARCD1 antibody [EPR23170-71] - BSA and Azide free (ab269463)
    Anti-SMARCD1 antibody [EPR23170-71] - BSA and Azide free (ab269463)

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