Recombinant
RabMAb

Recombinant Anti-mCherry antibody [EPR20579] (ab213511)

Overview

  • Product name

    Anti-mCherry antibody [EPR20579]
    See all mCherry primary antibodies
  • Description

    Rabbit monoclonal [EPR20579] to mCherry
  • Host species

    Rabbit
  • Tested applications

    Suitable for: ELISA, IP, Flow Cyt, WBmore details
    Unsuitable for: ICC/IF
  • Species reactivity

    Reacts with: Species independent
  • Immunogen

    Recombinant full length protein within mCherry aa 1 to the C-terminus. The exact sequence is proprietary. Anaplasma marginale.
    Database link: X5DSL3

  • Positive control

    • WB: 293T cells transfected with a His-tagged mCherry whole cell lysate. Flow Cyt: 293T cells transfected with a His-tagged mCherry. IP: 293T cells transfected with a His-tagged mCherry whole cell lysate.
  • 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

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
  • Storage buffer

    Preservative: 0.01% Sodium azide
    Constituents: 0.05% BSA, 40% Glycerol, PBS
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR20579
  • Isotype

    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab213511 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
ELISA Use at an assay dependent concentration.
IP 1/20.
Flow Cyt 1/400.
WB 1/1000. Detects a band of approximately 29 kDa (predicted molecular weight: 26 kDa).
  • Application notes
    Is unsuitable for ICC/IF.
  • Target

    • Relevance

      mCherry is derived from proteins originally isolated from Cnidarians (jelly fish, sea anemones and corals), and is used as a fluorescent tracer in trasfection and transgenic experiments. The prototype for these fluorescent proteins is Green Fluorescent Protein (GFP), which is a ~27kDa protein isolated originally from the jellyfish Aequoria victoria. The mCherry protein is derived from DsRed, a red fluorescent protein related to GFP isolated from so-called disc corals of the genus Discosoma.

    Images

    • Flow cytometric analysis of 4% paraformaldehyde-fixed 293T (human epithelial cell line from embryonic kidney transformed with large T antigen) transfected with a His-tagged mCherry cell line labeling mCherry with ab213511 at 1/400 dilution (right panel) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (left panel). Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/2000 dilution was used as the secondary antibody.

      mCherry positive population gives positive results as shown in right panel (Q2).

    • All lanes : Anti-mCherry antibody [EPR20579] (ab213511) at 1/1000 dilution

      Lane 1 : 293T (human epithelial cell line from embryonic kidney transformed with large T antigen) transfected with an empty expression vector whole cell lysate
      Lane 2 : 293T (human epithelial cell line from embryonic kidney transformed with large T antigen) transfected with a His-tagged mCherry whole cell lysate

      Lysates/proteins at 10 µg per lane.

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

      Developed using the ECL technique.

      Predicted band size: 26 kDa
      Observed band size: 29 kDa
      why is the actual band size different from the predicted?


      Exposure time: 5 seconds


      Blocking/Dilution buffer: 5% NFDM/TBST.

    • Anti-mCherry antibody [EPR20579] (ab213511) at 1/1000 dilution + Green fluorescent protein (GFP) at 0.01 µg

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

      Developed using the ECL technique.

      Predicted band size: 26 kDa


      Exposure time: 3 minutes


      Blocking/Dilution buffer: 5% NFDM/TBST.

      Green fluorescent protein (GFP) contains aa1-238 with His-tag.

    • mCherry was immunoprecipitated from 0.35 mg of 293T (human epithelial cell line from embryonic kidney transformed with large T antigen) transfected with a His-tagged mCherry whole cell lysate with ab213511 at 1/20 dilution. Western blot was performed from the immunoprecipitate using ab213511 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10,000 dilution.

      Lane 1:  293T transfected with a His-tagged mCherry whole cell lysate 10 µg (Input). 

      Lane 2: ab213511 IP in 293T transfected with a His-tagged mCherry whole cell lysate. 

      Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab213511 in 293T transfected with a His-tagged mCherry whole cell lysate.

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

      Exposure time: 10 seconds.

    References

    This product has been referenced in:

    • Williams JL  et al. Conditional Synaptic Vesicle Markers for Drosophila. G3 (Bethesda) 9:737-748 (2019). Read more (PubMed: 30635441) »
    • Flores BN  et al. An Intramolecular Salt Bridge Linking TDP43 RNA Binding, Protein Stability, and TDP43-Dependent Neurodegeneration. Cell Rep 27:1133-1150.e8 (2019). Read more (PubMed: 31018129) »
    See all 2 Publications for this product

    Customer reviews and Q&As

    1-5 of 5 Abreviews or Q&A

    Application
    Immunoprecipitation
    Sample
    Mouse Cell lysate - whole cell (MEF cells)
    Total protein in input
    500 µg
    Immuno-precipitation step
    Protein A/G
    Specification
    MEF cells

    Abcam user community

    Verified customer

    Submitted Mar 04 2019

    Application
    Immunoprecipitation
    Sample
    Human Cell lysate - whole cell (293T cells)
    Total protein in input
    500 µg
    Immuno-precipitation step
    Protein A/G
    Specification
    293T cells

    Abcam user community

    Verified customer

    Submitted Feb 12 2019

    Application
    Western blot
    Sample
    Mouse Cell lysate - whole cell (MEF cells)
    Gel Running Conditions
    Reduced Denaturing (4-12% Gradient Gel)
    Loading amount
    50 µg
    Specification
    MEF cells
    Blocking step
    Licor Blocking Buffer as blocking agent for 30 minute(s) · Concentration: 100% · Temperature: 25°C

    Abcam user community

    Verified customer

    Submitted Jan 24 2019

    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (293T cells)
    Gel Running Conditions
    Reduced Denaturing (4-12% Gradient Gel)
    Loading amount
    50 µg
    Specification
    293T cells
    Blocking step
    Licor Blocking Buffer as blocking agent for 30 minute(s) · Concentration: 100% · Temperature: 25°C

    Abcam user community

    Verified customer

    Submitted Jan 16 2019

    This product is known to not work in this application or species.
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Mouse Cell (Tumor Cells Engineered to Express mCherry)
    Permeabilization
    No
    Specification
    Tumor Cells Engineered to Express mCherry
    Blocking step
    10% Donkey Serum in 3% BSA/PBS as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 26°C
    Fixative
    Paraformaldehyde

    Dr. Francesco Elia Marino

    Verified customer

    Submitted Sep 21 2018

    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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