Recombinant

Anti-Mouse IgG1 VHH Single Domain Antibody (HRP) (isotype specific) (ab193651)

Overview

  • Product name
    Anti-Mouse IgG1 VHH Single Domain Antibody (HRP) (isotype specific)
  • Target species
    Mouse
  • Tested applications
    Suitable for: WB, ELISA, IHCmore details
  • Immunogen

    Full length protein corresponding to Mouse IgG1.

  • Conjugation
    HRP

Properties

  • Form
    Liquid
  • Storage instructions
    Shipped at 4°C. Store at 4°C (stable for up to 12 months). Store at -20°C long term. Avoid freeze / thaw cycle.
  • Storage buffer
    pH: 7.40
    Preservative: 0.1% Proclin
    Constituents: PBS, Sodium chloride, Sodium phosphate, 30% Glycerol, 1% BSA
  • Concentration information loading...
  • Purity
    Purified via His tag
  • Purification notes
    This product is a recombinant protein produced in E. coli.
  • Clonality
    Monoclonal
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab193651 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 Use at an assay dependent concentration.
ELISA Use at an assay dependent concentration.
IHC Use at an assay dependent concentration.

Images

  • All lanes : Anti-beta Actin antibody [mAbcam 8226] - Loading Control (ab8226) at 1 µg

    Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 2 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Anti-Mouse IgG1 VHH Single Domain (HRP) (isotype specific) (ab193651) at 0.1 µg/ml

    Developed using the ECL technique.

    Performed under reducing conditions.

    Observed band size: 42 kDa (why is the actual band size different from the predicted?)


    Exposure time: 5 seconds


    This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 2% Bovine Serum Albumin before being incubated with ab8226 overnight at 4°C. Antibody binding was detected using an anti-mouse IgG1 VHH single domain antibody conjugated to HRP (ab193651), and visualised using ECL development solution ab133406.

  • All lanes : Anti-GAPDH antibody [GA1R] (ab125247) at 2 µg/ml

    Lane 1 : HeLa Whole Cell Lysate
    Lane 2 : NIH 3T3 Whole Cell Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Anti-Mouse IgG1 VHH Single Domain (HRP) (isotype specific) (ab193651) at 0.1 µg/ml

    Developed using the ECL technique.

    Performed under reducing conditions.
  • IHC image of Anti-Mouse IgG1 VHH Single Domain Antibody (HRP) (isotype specific) (ab193651) staining in formalin fixed paraffin embedded normal human colon tissue section.

    The section was dewaxed and then pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6) in a Dako Pascal pressure cooker using the standard factory-set regime. Non-specific protein-protein interactions were then blocked using in TBS containing 0.025% (v/v) Triton X-100, 0.3M (w/v) glycine and 3% (w/v) BSA for 1h at room temperature. The section was then incubated with mouse IgG1 monoclonal antibody to Histone H1.0 (ab11080, 2.5µg/ml) in TBS containing 0.025% (v/v) Triton X-100 and 3% (w/v) BSA overnight at +4°C. Endogenous peroxidases were quenched using 1.6% (v/v) hydrogen peroxide in TBS containing 0.025% (v/v) Triton X-100 for 30 minutes at room temperature, with agitation. The secondary antibody, Anti-Mouse IgG1 VHH Single Domain Antibody (HRP) (isotype specific) (ab193651, 0.05µg/ml) was then applied for 1 hour at room temperature in TBS containing 0.025% (v/v) Triton X-100 and 3% (w/v) BSA before being developed for 10 minutes at room temperature using Steady DAB/Plus (ab103723). The section was then counterstained with hematoxylin and mounted with DPX.

    The negative control (secondary antibody only, no primary) inset shows no staining, demonstrating secondary antibody specificity.

    For other IHC staining systems (automated and non-automated), customers should optimize variable parameters such as antigen retrieval conditions, antibody concentrations and incubation times.

References

This product has been referenced in:
  • Wang Z  et al. Celastrol inhibits migration and invasion through blocking the NF-?B pathway in ovarian cancer cells. Exp Ther Med 14:819-824 (2017). Read more (PubMed: 28673005) »
  • Liu JY  et al. MicroRNA-153 inhibits the proliferation and invasion of human laryngeal squamous cell carcinoma by targeting KLF5. Exp Ther Med 11:2503-2508 (2016). WB . Read more (PubMed: 27284339) »

See all 5 Publications for this product

Customer reviews and Q&As

Application
Western blot
Experimental conditions:

MCF-7 whole cell lysates extracted with RIPA buffer.

10% SDS-PAGE, 50,000 cells loaded into wells.

Blocking 1h with 5% milk in TBS at RT.

Incubation with primary anti-alpha-Tubulin overnight at 4ºC.

Incubation with secondary antibody 0.1ug/ml in 5% milk for 1h at RT.

Developed with ECL for 5 minutes.

Results:
No nonspecific binding. No background. Clear bands.
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Submitted Jan 06 2015

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

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