Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-MSH6 antibody [EPR3945] - BSA and Azide free (ab214454)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR3945] to MSH6 - BSA and Azide free
  • Suitable for: ICC, WB, IHC-P
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-MSH6 antibody [EPR3945] - BSA and Azide free
    See all MSH6 primary antibodies

  • Description

    Rabbit monoclonal [EPR3945] to MSH6 - BSA and Azide free

  • Host species

    Rabbit

  • Tested applications

    Suitable for: ICC, WB, IHC-Pmore details

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

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

  • Positive control

    • WB: A431, HeLa, SW480. HAP1, and HEK-293 cell lysates IHC-P: Human colonic adenocarcinoma tissue ICC/IF: HeLa cells

  • General notes

    Ab214454 is the carrier-free version of ab92471. 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

    ab214454 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.

    Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

    Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

    We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

    In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

    We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

    Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

    Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.

Properties

Images

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MSH6 antibody [EPR3945] - BSA and Azide free (ab214454)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MSH6 antibody [EPR3945] - BSA and Azide free (ab214454)

    Immunohistochemical staining of paraffin embedded human colon with unpurified ab92471 at a dilution of 1/500. A pre-diluted HRP polymer for rabbit/mouse IgG was used as the secondary antibody and the sample was counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab92471).

  • Western blot - Anti-MSH6 antibody [EPR3945] - BSA and Azide free (ab214454)
    Western blot - Anti-MSH6 antibody [EPR3945] - BSA and Azide free (ab214454)
    All lanes : Anti-MSH6 antibody [EPR3945] (ab92471) at 1/1000 dilution

    Lane 1 : Wild-type HeLa cell lysate
    Lane 2 : MSH6 knockout HeLa cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 163 kDa
    Observed band size: 160 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 (ab92471).

      Lanes 1- 2: Merged signal (red and green). Green - ab92471 observed at 160 kDa. Red - Anti-Vinculin antibody [VIN-54] observed at 124 kDa.

     ab92471 was shown to react with MSH6 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab255410 (knockout cell lysate ab263763) was used. Wild-type HeLa and MSH6 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. ab92471 and Anti-Vinculin antibody [VIN-54] 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-MSH6 antibody [EPR3945] - BSA and Azide free (ab214454)
    Immunocytochemistry - Anti-MSH6 antibody [EPR3945] - BSA and Azide free (ab214454)

    ab92471 staining MSH6 in wild-type HAP1 cells (top panel) and MSH6 knockout HAP1 cells (bottom panel). The cells were fixed with 4% formaldehyde (10min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab92471 at 1μg/ml and ab195889 at 1/250 dilution (shown in pseudocolour red) overnight at +4°C, followed by a further incubation at room temperature for 1h with a goat secondary antibody to Rabbit IgG (Alexa Fluor® 488) (ab150081) at 2 μg/ml (shown in green). Nuclear DNA was labelled in blue with DAPI.

    Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab92471).

  • Western blot - Anti-MSH6 antibody [EPR3945] - BSA and Azide free (ab214454)
    Western blot - Anti-MSH6 antibody [EPR3945] - BSA and Azide free (ab214454)
    All lanes : Anti-MSH6 antibody [EPR3945] (ab92471) at 1/1000 dilution

    Lane 1 : Wild-type HAP1 cell lysate
    Lane 2 : MSH6 knockout HAP1 cell lysate
    Lane 3 : HeLa cell lysate
    Lane 4 : A431 cell lysate

    Lysates/proteins at 20 µg per lane.

    Predicted band size: 163 kDa



    This data was developed using the same antibody clone in a different buffer formulation (ab92471).

    Lanes 1 - 4: Merged signal (red and green). Green - ab92471 observed at 160 kDa. Red - loading control, ab18058, observed at 124 kDa.


    ab92471 was shown to specifically react with MSH6 in wild-type HAP1 cells. No band was observed when MSH6 knockout samples were used. Wild-type and MSH6 knockout samples were subjected to SDS-PAGE. ab92471 and ab18058 (loading control to Vinculin) were diluted at 1/1000 and 1/10 000 respectively and incubated overnight at 4°C. 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 1/10 000 dilution for 1 h at room temperature before imaging.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MSH6 antibody [EPR3945] - BSA and Azide free (ab214454)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MSH6 antibody [EPR3945] - BSA and Azide free (ab214454)

    Immunohistochemical staining of paraffin embedded rat liver with purified ab92471 at a dilution of 1/500. A pre-diluted HRP polymer for rabbit/mouse IgG was used as the secondary antibody and the sample was counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab92471).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MSH6 antibody [EPR3945] - BSA and Azide free (ab214454)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MSH6 antibody [EPR3945] - BSA and Azide free (ab214454)

    Immunohistochemical staining of paraffin embedded rat liver with unpurified ab92471 at a dilution of 1/150. A pre-diluted HRP polymer for rabbit/mouse IgG was used as the secondary antibody and the sample was counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab92471).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MSH6 antibody [EPR3945] - BSA and Azide free (ab214454)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MSH6 antibody [EPR3945] - BSA and Azide free (ab214454)

    Immunohistochemical staining of paraffin embedded human colon with unpurified ab92471 at a dilution of 1/150. A pre-diluted HRP polymer for rabbit/mouse IgG was used as the secondary antibody and the sample was counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab92471).

  • Immunocytochemistry - Anti-MSH6 antibody [EPR3945] - BSA and Azide free (ab214454)
    Immunocytochemistry - Anti-MSH6 antibody [EPR3945] - BSA and Azide free (ab214454)

    ab92471 staining MSH6 in HeLa cells. The cells were fixed with 4% formaldehyde (10min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab92471 at 1μg/ml and ab195889 at 1/250 dilution (shown in pseudocolour red) overnight at +4°C, followed by a further incubation at room temperature for 1h with a goat secondary antibody to Rabbit IgG (Alexa Fluor® 488) (ab150081) at 2 μg/ml (shown in green). Nuclear DNA was labelled in blue with DAPI.

    Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab92471).

  • Immunocytochemistry - Anti-MSH6 antibody [EPR3945] - BSA and Azide free (ab214454)
    Immunocytochemistry - Anti-MSH6 antibody [EPR3945] - BSA and Azide free (ab214454)

    Clone EPR3945 (ab214454) has been successfully conjugated by Abcam. This image was generated using Anti-MSH6 antibody [EPR3945] (Alexa Fluor® 647). Please refer to ab198334 for protocol details.

    ab198334 staining MSH6 in HeLa cells. The cells were fixed with 4% formaldehyde (10 min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab198334 at 1/100 dilution (shown in red) and ab195887, Mouse monoclonal to alpha Tubulin (Alexa Fluor® 488), at 1/250 dilution (shown in green). Nuclear DNA was labelled with DAPI (shown in blue).

    Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

    This product also gave a positive signal under the same testing conditions in HeLa cells fixed with 100% methanol (5min).

  • Immunocytochemistry - Anti-MSH6 antibody [EPR3945] - BSA and Azide free (ab214454)
    Immunocytochemistry - Anti-MSH6 antibody [EPR3945] - BSA and Azide free (ab214454) This image is courtesy of an Abreview submitted by Kirk McManus.

    Unpurified ab92471 (1/500) staining MSH6 in asynchronous HeLa cells (green). Cells were fixed in paraformaldehyde, permeabilised with 0.5% Triton X100/PBS and counterstained with DAPI in order to highlight the nucleus (red). For further experimental details please see abreview.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab92471).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MSH6 antibody [EPR3945] - BSA and Azide free (ab214454)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MSH6 antibody [EPR3945] - BSA and Azide free (ab214454)

    Unpurified ab92471, at a 1/100 dilution, detecting MSH6 in paraffin embedded Human colonic adenocarcinoma tissue by immunohistochemistry. Detection used HRP conjugated anti rabbit antibody.

    Perform heat mediated antigen retrieval 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 (ab92471).

  • OI-RD Scanning - Anti-MSH6 antibody [EPR3945] - BSA and Azide free (ab214454)
    OI-RD Scanning - Anti-MSH6 antibody [EPR3945] - BSA and Azide free (ab214454)
    Equilibrium disassociation constant (KD)
    Learn more about KD

    Click here to learn more about KD

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab92471).

  • Anti-MSH6 antibody [EPR3945] - BSA and Azide free (ab214454)
    Anti-MSH6 antibody [EPR3945] - BSA and Azide free (ab214454)

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