Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-MeCP2 antibody [EPR23201-3] - BSA and Azide free (ab273059)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR23201-3] to MeCP2 - BSA and Azide free
  • Suitable for: WB, ICC/IF, Flow Cyt, IHC-P
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-MeCP2 antibody [EPR23201-3] - BSA and Azide free
    See all MeCP2 primary antibodies

  • Description

    Rabbit monoclonal [EPR23201-3] to MeCP2 - BSA and Azide free

  • Host species

    Rabbit

  • Specificity

    This antibody did not work in IHC in human brain samples.

  • Tested Applications & Species

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

  • Immunogen

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

  • Positive control

    • WB: HAP1, HepG2, SH-SY5Y, HEK-293T, Neuro-2a cell lysates; Mouse brain and hippocampus tissue lysate; Rat brain and liver tissue lysates; Human brain, hippocampus and liver lysates. IHC-P: Mouse lung, Mouse testis, Rat cerebrum, Rat lung and Human kidney, Human astrocytoma tissues. ICC/IF: SH-SY5Y and Neuro-2a cells. Flow Cyt: SH-SY5Y and Neuro-2a cells.

  • General notes

    ab273059 is the carrier-free version of ab253197. 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

    This product 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-MeCP2 antibody [EPR23201-3] - BSA and Azide free (ab273059)
    Western blot - Anti-MeCP2 antibody [EPR23201-3] - BSA and Azide free (ab273059)
    All lanes : Anti-MeCP2 antibody [EPR23201-3] (ab253197) at 1/1000 dilution

    Lane 1 : Wild-type HAP1 cell lysate
    Lane 2 : MECP2 knockout HAP1 cell lysate
    Lane 3 : SH-SY5Y cell lysate
    Lane 4 : Mouse Brain cell lysate
    Lane 5 : HepG2 cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 52 kDa
    Observed band size: 68-70 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 (ab253197).

    Lanes 1 - 5: Merged signal (red and green). Green - ab253197 observed at 68-70 kDa. Red - loading control, ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.

    ab253197 was shown to react with MeCP2 in HAP1 wild-type cells in western blot. Loss of signal was observed when MECP2 knockout sample was used. Wild-type and MECP2 HAP1 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 5% milk in TBS-T (0.1% Tween®) before incubation with ab253197 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MeCP2 antibody [EPR23201-3] - BSA and Azide free (ab273059)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MeCP2 antibody [EPR23201-3] - BSA and Azide free (ab273059)

    Immunohistochemical analysis of paraffin-embedded Human astrocytoma tissue labeling MeCP2 with ab253197 at 1/4000 dilution (0.158 ug/ml) dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining on human astrocytoma (PMID: 11809720). The section was incubated with ab253197 for 10 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

    Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

    Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab253197).

  • Immunocytochemistry/ Immunofluorescence - Anti-MeCP2 antibody [EPR23201-3] - BSA and Azide free (ab273059)
    Immunocytochemistry/ Immunofluorescence - Anti-MeCP2 antibody [EPR23201-3] - BSA and Azide free (ab273059)
    Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized SH-SY5Y cells labelling MeCP2 with ab253197 at 1/500 dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 2 ug/ml dilution (Green). Confocal image showing nuclear staining in SH-SY5Y cells. Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5 dilution (Red). The Nuclear counterstain was DAPI (Blue).

    Secondary antibody only control: Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1000 2 ug/ml dilution.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab253197).

  • Flow Cytometry - Anti-MeCP2 antibody [EPR23201-3] - BSA and Azide free (ab273059)
    Flow Cytometry - Anti-MeCP2 antibody [EPR23201-3] - BSA and Azide free (ab273059)
    Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized SH-SY5Y (Human neuroblastoma epithelial cell) cells labelling MeCP2 with ab253197 at 1/600 dilution (0.1ug) (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 (ab253197).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MeCP2 antibody [EPR23201-3] - BSA and Azide free (ab273059)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MeCP2 antibody [EPR23201-3] - BSA and Azide free (ab273059)

    Immunohistochemical analysis of paraffin-embedded Mouse testis tissue labeling MeCP2 with ab253197 at 1/4000 dilution (0.158 ug/ml) dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining on mouse testis (PMID: 11809720). The section was incubated with ab253197 for 10 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

    Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

    Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab253197).

  • Immunocytochemistry/ Immunofluorescence - Anti-MeCP2 antibody [EPR23201-3] - BSA and Azide free (ab273059)
    Immunocytochemistry/ Immunofluorescence - Anti-MeCP2 antibody [EPR23201-3] - BSA and Azide free (ab273059)
    Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized Neuro-2a cells labelling MeCP2 with ab253197 at 1/500 dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 2 ug/ml 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 2.5 dilution (Red). The Nuclear counterstain was DAPI (Blue).

    Secondary antibody only control: Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1000 2 ug/ml dilution.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab253197).

  • Flow Cytometry - Anti-MeCP2 antibody [EPR23201-3] - BSA and Azide free (ab273059)
    Flow Cytometry - Anti-MeCP2 antibody [EPR23201-3] - BSA and Azide free (ab273059)
    Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Neuro-2a (Mouse neuroblastoma neuroblast) cells labelling MeCP2 with ab253197 at 1/600 dilution (0.1ug) (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 (ab253197).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MeCP2 antibody [EPR23201-3] - BSA and Azide free (ab273059)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MeCP2 antibody [EPR23201-3] - BSA and Azide free (ab273059)

    Immunohistochemical analysis of paraffin-embedded Rat lung tissue labeling MeCP2 with ab253197 at 1/4000 dilution (0.158 ug/ml) dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining on rat lung (PMID: 11809720). The section was incubated with ab253197 for 10 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

    Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

    Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab253197).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MeCP2 antibody [EPR23201-3] - BSA and Azide free (ab273059)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MeCP2 antibody [EPR23201-3] - BSA and Azide free (ab273059)

    Immunohistochemical analysis of paraffin-embedded Human kidney tissue labeling MeCP2 with ab253197 at 1/4000 dilution (0.158 ug/ml) dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining on human kidney (PMID: 11809720). The section was incubated with ab253197 for 10 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

    Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

    Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab253197).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MeCP2 antibody [EPR23201-3] - BSA and Azide free (ab273059)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MeCP2 antibody [EPR23201-3] - BSA and Azide free (ab273059)

    Immunohistochemical analysis of paraffin-embedded Rat cerebrum tissue labeling MeCP2 with ab253197 at 1/4000 dilution (0.158 ug/ml) dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining on rat cerebrum (PMID: 11809720). The section was incubated with ab253197 for 10 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

    Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

    Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab253197).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MeCP2 antibody [EPR23201-3] - BSA and Azide free (ab273059)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MeCP2 antibody [EPR23201-3] - BSA and Azide free (ab273059)

    Immunohistochemical analysis of paraffin-embedded Mouse lung tissue labeling MeCP2 with ab253197 at 1/4000 dilution (0.158 ug/ml) dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining on mouse lung (PMID: 11809720). The section was incubated with ab253197 for 10 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

    Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

    Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab253197).

  • Anti-MeCP2 antibody [EPR23201-3] - BSA and Azide free (ab273059)
    Anti-MeCP2 antibody [EPR23201-3] - BSA and Azide free (ab273059)

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