Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-SIRT5 antibody [EPR23787-116] - BSA and Azide free (ab275031)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR23787-116] to SIRT5 - BSA and Azide free
  • Suitable for: WB, Flow Cyt, IHC-P, IP
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-SIRT5 antibody [EPR23787-116] - BSA and Azide free
    See all SIRT5 primary antibodies

  • Description

    Rabbit monoclonal [EPR23787-116] to SIRT5 - BSA and Azide free

  • Host species

    Rabbit

  • Tested Applications & Species

    Application Species
    Flow Cyt
    Human
    IHC-P
    Mouse
    IP
    Human
    WB
    Human
    See all applications and species data

  • Immunogen

    Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: HepG2, Huh7, HeLa, HEK-293T, Hepa1-6, RAW 264.7 and C6 whole cell lysates; Human liver and heart tissue lysates; Mouse liver and heart tissue lysates; Rat liver and tissue lysates. IHC-P: Human liver and ovarian adenocarcinoma tissue; Mouse liver tissue; Rat liver tissue. Flow cyt: HepG2 and Hepa1-6 cells. IP: HepG2 whole cell lysate; Mouse liver tissue lysate.

  • General notes

    ab275031 is the carrier-free version of ab259967. 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-SIRT5 antibody [EPR23787-116] - BSA and Azide free (ab275031)
    Western blot - Anti-SIRT5 antibody [EPR23787-116] - BSA and Azide free (ab275031)
    All lanes : Anti-SIRT5 antibody [EPR23787-116] (ab259967) at 1/1000 dilution

    Lane 1 : Mouse liver tissue lysate
    Lane 2 : Mouse heart tissue lysate
    Lane 3 : Rat liver tissue lysate
    Lane 4 : Rat heart tissue lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/20000 dilution

    Predicted band size: 33 kDa
    Observed band size: 30 kDa
    why is the actual band size different from the predicted?



    This data was developed using ab259967, the same antibody clone in a different buffer formulation.

    Blocking and dilution buffer: 5% NFDM/TBST.

    Exposure time: 48 seconds.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SIRT5 antibody [EPR23787-116] - BSA and Azide free (ab275031)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SIRT5 antibody [EPR23787-116] - BSA and Azide free (ab275031)

    This data was developed using ab259967, the same antibody clone in a different buffer formulation.

    Immunohistochemical analysis of paraffin-embedded Mouse liver tissue labeling SIRT5 with ab259967 at 1/100 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on mouse liver (PMID 29491006). 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 using ab93684 (Tris/EDTA buffer, pH 9.0).

  • Immunoprecipitation - Anti-SIRT5 antibody [EPR23787-116] - BSA and Azide free (ab275031)
    Immunoprecipitation - Anti-SIRT5 antibody [EPR23787-116] - BSA and Azide free (ab275031)

    This data was developed using ab259967, the same antibody clone in a different buffer formulation.

    SIRT5 was immunoprecipitated from 0.35 mg Mouse liver tissue lysate with ab259967 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab259967 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

    Lane 1: Mouse liver tissue lysate 10 ug

    Lane 2: ab259967 IP in Mouse liver tissue lysate

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab259967 in mouse liver tissue lysate

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: 3 minutes.

     

  • Western blot - Anti-SIRT5 antibody [EPR23787-116] - BSA and Azide free (ab275031)
    Western blot - Anti-SIRT5 antibody [EPR23787-116] - BSA and Azide free (ab275031)
    All lanes : Anti-SIRT5 antibody [EPR23787-116] (ab259967) at 1/1000 dilution

    Lane 1 : HepG2 (human hepatocellular carcinoma epithelial cell) whole cell lysate
    Lane 2 : Huh7 (human hepatocellular carcinoma epithelial cell) whole cell lysate
    Lane 3 : HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate
    Lane 4 : HEK-293T (human embryonic kidney epithelial cell) whole cell lysate
    Lane 5 : Hepa1-6 (mouse hepatoma epithelial cell) whole cell lysate
    Lane 6 : RAW 264.7 (mouse abelson murine leukemia virus-induced tumor macrophage) whole cell lysate
    Lane 7 : C6 (rat glial tumor glial cell) whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/20000 dilution

    Predicted band size: 33 kDa
    Observed band size: 30 kDa why is the actual band size different from the predicted?



    This data was developed using ab259967, the same antibody clone in a different buffer formulation.

    Blocking and dilution buffer: 5% NFDM/TBST.

    Exposure time: 3 minutes.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SIRT5 antibody [EPR23787-116] - BSA and Azide free (ab275031)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SIRT5 antibody [EPR23787-116] - BSA and Azide free (ab275031)

    This data was developed using ab259967, the same antibody clone in a different buffer formulation.

    Immunohistochemical analysis of paraffin-embedded Human liver tissue labeling SIRT5 with ab259967 at 1/100 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on human liver (PMID 29491006). 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 using ab93684 (Tris/EDTA buffer, pH 9.0).

  • Immunoprecipitation - Anti-SIRT5 antibody [EPR23787-116] - BSA and Azide free (ab275031)
    Immunoprecipitation - Anti-SIRT5 antibody [EPR23787-116] - BSA and Azide free (ab275031)

    This data was developed using ab259967, the same antibody clone in a different buffer formulation.

    SIRT5 was immunoprecipitated from 0.35 mg HepG2 (human hepatocellular carcinoma epithelial cell) whole cell lysate with ab259967 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab259967 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

    Lane 1: HepG2 (human hepatocellular carcinoma epithelial cell) whole cell lysate 10 ug

    Lane 2: ab259967 IP in HepG2 whole cell lysate

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab259967 in HepG2 whole cell lysate

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: 3 minutes.

     

  • Flow Cytometry - Anti-SIRT5 antibody [EPR23787-116] - BSA and Azide free (ab275031)
    Flow Cytometry - Anti-SIRT5 antibody [EPR23787-116] - BSA and Azide free (ab275031)

    This data was developed using ab259967, the same antibody clone in a different buffer formulation.

    Flow cytometric analysis of 4% paraformaldehyde fixed, 90% methanol permeabilized Hepa1-6 (Mouse hepatoma epithelial cell) cells labelling SIRT5 with ab259967 at 1/50 dilution (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.

  • Western blot - Anti-SIRT5 antibody [EPR23787-116] - BSA and Azide free (ab275031)
    Western blot - Anti-SIRT5 antibody [EPR23787-116] - BSA and Azide free (ab275031)
    All lanes : Anti-SIRT5 antibody [EPR23787-116] (ab259967) at 1/1000 dilution

    Lane 1 : Human liver tissue lysate
    Lane 2 : Human heart tissue lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : VeriBlot for IP secondary antibody(HRP)(ab131366) at 1/1000 dilution

    Predicted band size: 33 kDa
    Observed band size: 30 kDa why is the actual band size different from the predicted?



    This data was developed using ab259967, the same antibody clone in a different buffer formulation.

    Blocking and dilution buffer: 5% NFDM/TBST.

    Exposure time: 3 minutes.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SIRT5 antibody [EPR23787-116] - BSA and Azide free (ab275031)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SIRT5 antibody [EPR23787-116] - BSA and Azide free (ab275031)

    This data was developed using ab259967, the same antibody clone in a different buffer formulation.

    Immunohistochemical analysis of paraffin-embedded Rat liver tissue labeling SIRT5 with ab259967 at 1/100 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on rat liver (PMID 29491006). 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 using ab93684 (Tris/EDTA buffer, pH 9.0).

  • Flow Cytometry - Anti-SIRT5 antibody [EPR23787-116] - BSA and Azide free (ab275031)
    Flow Cytometry - Anti-SIRT5 antibody [EPR23787-116] - BSA and Azide free (ab275031)

    This data was developed using ab259967, the same antibody clone in a different buffer formulation.

    Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HepG2 (Human hepatocellular carcinoma epithelial cell) cells labelling SIRT5 with ab259967 at 1/50 dilution (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.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SIRT5 antibody [EPR23787-116] - BSA and Azide free (ab275031)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SIRT5 antibody [EPR23787-116] - BSA and Azide free (ab275031)

    This data was developed using ab259967, the same antibody clone in a different buffer formulation.

    Immunohistochemical analysis of paraffin-embedded Human ovarian adenocarcinoma tissue labeling SIRT5 with ab259967 at 1/100 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on human ovarian adenocarcinoma (PMID 31456942). 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 using ab93684 (Tris/EDTA buffer, pH 9.0).

  • Anti-SIRT5 antibody [EPR23787-116] - BSA and Azide free (ab275031)
    Anti-SIRT5 antibody [EPR23787-116] - BSA and Azide free (ab275031)

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