Recombinant
RabMAb

Recombinant Anti-BHMT + BHMT2 antibody [EPR20822] (ab213491)

Overview

  • Product name

    Anti-BHMT + BHMT2 antibody [EPR20822]
  • Description

    Rabbit monoclonal [EPR20822] to BHMT + BHMT2
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IHC-Pmore details
  • Species reactivity

    Reacts with: Rat, Human
  • Immunogen

    Recombinant fragment within Human BHMT2 aa 1 to the C-terminus. The exact sequence is proprietary.
    Database link: Q9H2M3

  • Positive control

    • WB: His-tagged human BHMT2 and BHMT recombinant proteins; Human fetal liver and fetal kidney tissue lysate; Rat liver lysate. IHC-P: Human kidney and liver tissues; Rat liver tissue.
  • General notes

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab213491 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
WB 1/1000. Predicted molecular weight: 40 kDa.
IHC-P 1/20000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

IHC note: 1:20,000, recommend human and rat. BHMT2 involved in the regulation of homocysteine metabolism. It’s mainly expressed on liver and kidney and its subcellular location was cytoplasm. IHC results showed specific staining on liver and kidney of human and liver, mouse kidney was very weak staining and testis leydig cells were non-specific staining. So IHC recommend human and rat.

Note: It can show nuclear staining if working at higher concentration.

Target

Images

  • Immunohistochemical analysis of paraffin-embedded human kidney tissue labeling BHMT + BHMT2 with ab213491 at 1/20,000 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Cytoplasmic staining on human kidney tissue is observed (PMID: 9281325; PMID: 26592251). Counter stained with hematoxylin. Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

    Perform heat-mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

    IHC note: 1:20,000, recommend human and rat. BHMT2 involved in the regulation of homocysteine metabolism. It’s mainly expressed on liver and kidney and its subcellular location was cytoplasm. IHC results showed specific staining on liver and kidney of human and liver, mouse kidney was very weak staining and testis leydig cells were non-specific staining. So IHC recommend human and rat.

    Note: It can show nuclear staining if working at higher concentration.

  • Immunohistochemical analysis of paraffin-embedded human liver tissue labeling BHMT + BHMT2 with ab213491 at a 1/20,000 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Cytoplasmic staining on human liver is observed (PMID: 9281325; PMID: 26592251). Counter stained with hematoxylin. Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

    Perform heat-mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

  • Immunohistochemical analysis of paraffin-embedded rat liver tissue labeling BHMT + BHMT2 with ab213491 at 1/20,000 dilution followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Cytoplasmic staining on rat liver is observed (PMID: 9281325; PMID: 26592251).counterstained with hematoxylin. Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

    Perform heat-mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

  • All lanes : Anti-BHMT + BHMT2 antibody [EPR20822] (ab213491) at 1/1000 dilution

    Lane 1 : Recombinant Human BHMT2 protein (denatured) (ab180277)
    Lane 2 : Recombinant Human BHMT protein (ab51280)

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)

    Predicted band size: 40 kDa



    Blocking/Diluting buffer and concentration: 5% NFDM/TBST

    Lane 1 and 2 samples loaded at 10 ng.

    Exposure time: 58 seconds.

    The blot was developed on a BIO-RAD® ChemiDoc™ MP instrument.

  • All lanes : Anti-BHMT + BHMT2 antibody [EPR20822] (ab213491) at 1/1000 dilution

    Lane 1 : Human fetal liver tissue
    Lane 2 : Human fetal kidney tissue
    Lane 3 : Human fetal spleen tissue

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)

    Predicted band size: 40 kDa



    Blocking/Diluting buffer and concentration: 5% NFDM/TBST 

    Exposure time: 3 minutes

    BHMT and BHMT2 are mainly expressed in liver and kidney, negative in spleen and other tissues (PMID18457970; PMID: 11087663). The blot was developed on a BIO-RAD® ChemiDoc™ MP instrument.

  • Anti-BHMT + BHMT2 antibody [EPR20822] (ab213491) at 1/1000 dilution + Rat liver tissue at 20 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)

    Predicted band size: 40 kDa
    Observed band size: 45 kDa
    why is the actual band size different from the predicted?



    Blocking/Diluting buffer and concentration: 5% NFDM/TBST 

    Exposure time: 7 seconds.

    The blot was developed on a BIO-RAD® ChemiDoc™ MP instrument.

References

ab213491 has not yet been referenced specifically in any publications.

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