Recombinant
RabMAb

Recombinant Anti-Transglutaminase 2 antibody [EPR2956] (ab109121)

Rabbit recombinant monoclonal Transglutaminase 2 antibody [EPR2956]. Validated in WB and tested in Mouse, Rat, Human.

Overview

  • Product name

    Anti-Transglutaminase 2 antibody [EPR2956]
    See all Transglutaminase 2 primary antibodies
  • Description

    Rabbit monoclonal [EPR2956] to Transglutaminase 2
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WBmore details
    Unsuitable for: Flow Cyt,IHC-P or IP
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Human Transglutaminase 2 aa 350-450. The exact sequence is proprietary.

  • Positive control

    • ECV-304 and HUVEC cell lysates.
  • 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 ab109121 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 - 1/10000. Predicted molecular weight: 77 kDa.
  • Application notes
    Is unsuitable for Flow Cyt,IHC-P or IP.
  • Target

    • Function

      Catalyzes the cross-linking of proteins and the conjugation of polyamines to proteins.
    • Sequence similarities

      Belongs to the transglutaminase superfamily. Transglutaminase family.
    • Information by UniProt
    • Database links

    • Alternative names

      • ALPHA SUBUNIT antibody
      • C polypeptide antibody
      • EC 2.3.2.13 antibody
      • epididymis secretory protein Li 45 antibody
      • G alpha h antibody
      • G[a]h antibody
      • Gh CLASS G ALPHA h antibody
      • GNAH antibody
      • GNAH G PROTEIN antibody
      • H POLYPEPTIDE antibody
      • HEL-S-45 antibody
      • Protein glutamine gamma glutamyltransferase 2 antibody
      • Protein-glutamine gamma-glutamyltransferase 2 antibody
      • TG 2 antibody
      • TG(C) antibody
      • TG2 antibody
      • TGase C antibody
      • TGase H antibody
      • TGase-2 antibody
      • TgaseII antibody
      • TGC antibody
      • TGC GUANINE NUCLEOTIDE BINDING PROTEIN antibody
      • TGM2 antibody
      • TGM2_HUMAN antibody
      • Tissue transglutaminase antibody
      • Transglutaminase 2 antibody
      • Transglutaminase 2 C polypeptide antibody
      • Transglutaminase C antibody
      • Transglutaminase H antibody
      • Transglutaminase-2 antibody
      • tTG antibody
      • tTGas antibody
      see all

    Images

    • All lanes : Anti-Transglutaminase 2 antibody [EPR2956] (ab109121) at 1/1000 dilution

      Lane 1 : Mouse heart tissue lysate
      Lane 2 : Rat heart tissue lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

      Predicted band size: 77 kDa


      Exposure time: 3 minutes


      Blocking and diluting buffer: 5% NFDM/TBST.

    • All lanes : Anti-Transglutaminase 2 antibody [EPR2956] (ab109121) at 1/1000 dilution

      Lane 1 : ECV-304 cell lysates
      Lane 2 : HUVEC cell lysates

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : HRP labelled Goat anti-Rabbit at 1/2000 dilution

      Predicted band size: 77 kDa

    References

    ab109121 has not yet been referenced specifically in any publications.

    Customer reviews and Q&As

    Question

    Thank you very much for your reply, I forwarded the mail to the customer. She now explains her project and wants me to ask you if you have any ideas, or products which she might be able to use.   Here is her mail:   Thanks so much for finding out from the company for me. I have rodent mitochondria samples that I would like to use, and preferably I would like to test all 4 or 5 of the electron transport chain complexes - that is why I mentioned the 5x96-well plate assay as it seemed to me that this could test all 5 complexes. The reason I am interested in this product is to find an alternative to the Oxygraph method that measures oxygen consumption and electron transport chain complex functioning. I am however at the stage where I do not know whether all 5 of the complexes' activity will be altered with my intervention (an antiretroviral drug). I have looked through the database and discussed this with my promotor, and we suspect that as a first step we could try the MitoProfile Membrane Integrity Cocktail antibody as well as the ATPase/Complex V specific activity to get an idea of what is happening. I have previously ordered the MitoOxphos Complex I-V Cocktail (and will order it again soon - I require more), and I believe that this might give us better leverage as how to go forward with our hypothesis. My hypothesis is that one of the antiretroviral agents used in HIV/AIDS treatment can induce mitochondrial abnormalities, and studies have shown various effects ranging from changes to mitochondrial DNA, mitochondrial gene expression and membrane integrity. However my study is quite novel and we are unsure whether our experimental conditions would confirm previous findings, and if the drug could alter not only gene and protein expression levels of the complexes, but affect functioning with ATP production and structural integrity. You are welcome to pass this information on to your contact at Abcam - perhaps with this info they can advise more appropriately. I do however think that the ATPase specific activity assay might be a very good choice to later substantiate Oxygraph analyses.  

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    Answer

    Thank you for your reply. I have been in touch with our colleagues at Mitosciences who have kindly provided the following information: To answer your question directly, The antibody coating the plate supplied with Kit MTOX1 ab109903 does not cross-react with rat samples (it is human/bovine only).  If you would like to  test complex I activity in Rat mitochondria, it would be better to purchase kit ab109121.  Click here (or use the following: https://www.abcam.com/index.html?datasheet=109121). However, the drawback of kit ab109121 is that it is not rotenone sensitive as it is only the dehydrogenase activity of the enzyme and does not follow the complete reaction (dehydrogenase-ubiquinol reductase).  This is explained in detail in the protocol on the datasheet. To answer the complete question, we have had a substantial experience testing mitochondrial toxicity due to antiretrovirals.  It looks to me that you are treating animals with an antiretroviral compound for HIV?  The effect on mitochondria due to antiretroviral therapy varies tremendously depending on the regime used and to my knowledge the most prominent causes of mito toxicity are : (1) inhibition of chain elongation and/or exonuclease activity of the mtDNA polymerase γ, (2) inhibition of thymidine kinases, (3) oxidative stress, (4) impairment of the adenine nucleotide translocase and (5) reduction in mitochondrial membrane potential (Δψm).  The easiest and most economical way of looking at such a wide spread of potential effects in mitochondria is by testing cultured cells exposed to the compound with the following sets of kits: ab113849 ATP luminescent detection kit = check the scientific support tab as well as the protocol for further insight into mitochondrial toxicity testing.  This can test the mitochondrial bioenergetic state of the cell by comparing treatment in glucose vs. galactose based media. Click here (or use the following: https://www.abcam.com/index.html?datasheet=113849). ab113850 JC1 or ab113852 TMRE.  Both of these test membrane potential.  The protocol also has insights as to the effect of low membrane potential in the overall mitochondrial metabolism. Click here (or use the following: https://www.abcam.com/index.html?datasheet=113850). Click here (or use the following: https://www.abcam.com/index.html?datasheet=113852). ab113851 DCFDA = which measures ROS. Click here (or use the following: https://www.abcam.com/index.html?datasheet=113851). ab110217 and ab110216 Mitobiogenesis In-cell ELISA kits (colorimetric or IR) = These will measure the effect of a compound on the protein levels of a mitochondrial DNA encoded protein (typically affected by nucleoside reverse transcriptase inhibitors) in comparison to the protein levels of a nuclear DNA encoded protein (not affected by NRTIs).  Click here (or use the following: https://www.abcam.com/index.html?datasheet=110217). Click here (or use the following: https://www.abcam.com/index.html?datasheet=110216). The kits above mention can be used with any mammalian cell.  The mitobiogenesis kits can be used with human, rat, mouse or bovine cells.  The protocols explain in detail how to set the experiments.  If you haven’t tested the compound with the first basic acute tests on cells, I would recommend to do this.  It will help to narrow down the potential cause of toxicity or at least help have a viable hypothesis of what could be happening in the animal.  If the first four are normal (after acute treatment) I would suggest for to move ahead with the mitobiogenesis kit after treating the cells for at least 5 – 7 passages.  If they have a very good reason to suspect (due to the chemical structure of the compound) that the effect will be only on the mtDNA polymerase, then my suggestion is to go directly to the ab110217 to confirm the hypothesis. Once you have a reasonable hypothesis of the effect of the compound in-vitro, then choosing the test for in-vivo testing will be much easier.  i.e if the retroviral has an important effect on ROS production then they may want to follow up with antibodies that target ROS induced post-translational effects such as anti-nitrotyrosine, ab110282. Click here (or use the following: https://www.abcam.com/index.html?datasheet=110282). If galactose sensitizes the cells to compound toxicity as measured by the ATP luminescent assay, then very likely the compound is affecting the enzyme activity/assembly/levels of one of the complexes of the electron transport chain.  In this instance, then they will need to determine if it is a direct or indirect effect on the enzymes.  The MTOXC kits (5 kits) will test  whether the compound affects directly any of the enzymes using bovine mitochondria (Note that if a compound directly inhibits the transfer of electrons, by in-vitro testing, in bovine mitochondria, it will very likely affect the transfer in of electrons in other mammalian species such as rat).  MTOXC will test activity in the presence of compound (while the assay runs) The following MS kits: Complex I Enzyme Activity Microplate Assay Kit ab109721, Complex II Enzyme Activity Microplate Assay Kit ab109908, Complex IV Rodent Enzyme Activity Microplate Assay Kit ab109911 and ATP synthase Enzyme Activity Microplate Assay Kit ab109714,  all of which are rat reactive) will test indirect effect of the compound in the activity of enzymes (post-translational modifications, assembly defects) in tissues of treated animals.   ab109721 Click here (or use the following: https://www.abcam.com/index.html?datasheet=109721). ab109908 Click here (or use the following: https://www.abcam.com/index.html?datasheet=109908). ab109911 Click here (or use the following: https://www.abcam.com/index.html?datasheet=109911). ab109714 Click here (or use the following: https://www.abcam.com/index.html?datasheet=109714). These MS kits test activity of tissues or cell extracts after they have been exposed to an experimental condition.  For example if you test NRTI compounds with MTOXC, all results will be normal.  However if you test with the MS kits tissues from animals treated with NRTIs for a prolonged period of time, you will see decrease in activity.  You could use the following antibody cocktails: ab110413 MitoProfile® Total OXPHOS Rodent WB Antibody Cocktail  Click here (or use the following: https://www.abcam.com/index.html?datasheet=110413). ab110414 MitoProfile® Membrane Integrity WB Antibody Cocktail  Click here (or use the following: https://www.abcam.com/index.html?datasheet=110414). If you believe that the structural membrane integrity of the mitochondria (of treated animals) can have an important impact in complexes assembly and protein levels in specific compartments, then these cocktails may be a good choice.  Note that the ATP synthase Enzyme Activity kit ab109714 measures the reverse reactions (ATP hydrolysis) and not ATP synthesis, however the ATP hydrolysis is still oligomycin sensitive.  A decrease in oxygen consumption by the oxygraph could be due to effects in any of the complexes and not just ATPase.  I.e. rotenone affects oxygraph results, but it is a complex I specific inhibitor. So choosing this early one MS541 kit may lead them into a wild goose chase.  You would need to look at mitochondria in a more holistic way and not just as a measure of ATP synthesis.   I hope this information will be helpful to you. if you have any further questions, please do not hesitate to contact us.

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