Anti-HuB+HuC+HuD antibody [16A11C1D4] (ab176106)

Overview

  • Product name
    Anti-HuB+HuC+HuD antibody [16A11C1D4]
  • Description
    Mouse monoclonal [16A11C1D4] to HuB+HuC+HuD
  • Host species
    Mouse
  • Tested applications
    Suitable for: ELISA, ICC/IF, WBmore details
  • Species reactivity
    Reacts with: Human
    Does not react with: Mouse, Rat
  • Immunogen

    Synthetic peptide corresponding to Human HuB+HuC+HuD aa 233-244 conjugated to Keyhole Limpet Haemocyanin (KLH) (Cysteine residue).
    Sequence:

    QAQRFRLDNLLN


    Database link: Q12926

  • Positive control
    • HuC Human recombinant protein fragment His-tagged; HuD Human recombinant protein; SH-SY5Y cell lysate; SH-SY5Y cells.

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: 0.02% Sodium azide
    Constituent: 99% HEPES buffered saline
  • Concentration information loading...
  • Purification notes
    The purity of ab176106 is near homogeneity as judged by SDS-PAGE. The antibody was produced in vitro using hybridomas grown in serum-free medium, and then concentrated by chemical fractionation.
  • Clonality
    Monoclonal
  • Clone number
    16A11C1D4
  • Isotype
    IgG2b
  • Light chain type
    kappa

Applications

Our Abpromise guarantee covers the use of ab176106 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 a concentration of 1 µg/ml.

(or use at assay dependent concentration).

ICC/IF Use a concentration of 1 µg/ml.
WB Use a concentration of 1 µg/ml. Predicted molecular weight: 40, 42 kDa.

Images

  • All lanes : Anti-HuB+HuC+HuD antibody [16A11C1D4] (ab176106) at 1 µg/ml

    Lane 1 : Human HuC protein fragment (ab91911) at 0.1 µg
    Lane 2 : HuD human recombinant protein at 0.1 µg
    Lane 3 : SH-SY5Y cell lysate at 20 µg

    Secondary
    All lanes : Goat polyclonal to Mouse IgG - HRP at 1/5000 dilution

    Predicted band size: 40, 42 kDa



    Predicted MW (kDa): HuB/ELAVL2: 39.5 kDa, HuC/ELAVL3: 39.5kDa, HuD/ELAVL4: 41.7kDa

    Why the observed band size is different than the predicted band size: HuC protein (ab91911) is a fragment and has an N terminal His tag and is predicted to present at 24 kDa.

  • Immunocytochemistry with SH-SY5Y cells stained with ab176106 in red and DAPI in blue, as a counter stain.

    Fixation: Paraformaldehyde fixed (4%, 20 minutes).

    Permeabilization: Triton X-100 permeabilized (0.1%, 10 minutes).

    Blocking: 10% Goat Serum/PBS (1 hour).

    Primary: ab176106, 1 µg/ml (10% Goat Serum/PBS, 2 hours at room temperature or over night at 4°C).

    Secondary: Alexa 594 goat anti-mouse H+L used at a 1/1000 dilution in 1%Goat Serum/PBS for 1hour at RT.

    Washing: 3X 1% Goat Serum/PBS, 10 minutes per wash.

    DAPI: 2 uM in 1% Goat Serum/PBS, 10 minutes.

  • Specificity of antibody using the ELISA platform.

    Antibody concentration is at 1µg/mL the concentration of recombinant proteins is at 250ng/mL. ab176106 reacts specifically to HuC/ELAVL3 and HuD/ELAVL4 proteins. Reactivity with HuB protein is predicted by the immunogen sequence similarity.

References

ab176106 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Monkey Cell (Kidney)
Permeabilization
Yes - 0.1% triton x-100
Specification
Kidney
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 37°C
Fixative
Paraformaldehyde
Username

Abcam user community

Verified customer

Submitted Jun 10 2016

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (Brain)
Permeabilization
No
Specification
Brain
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 37°C
Fixative
Paraformaldehyde
Username

Abcam user community

Verified customer

Submitted Mar 04 2016

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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