Overview

  • Product name
    Rabbit Loading Control Antibody Panel
    See all Loading Control Panel kits
  • Product overview

    Ab199712 is a sampler pack of loading control antibodies and a Goat Anti-Rabbit IgG secondary antibody conjugated to HRP.
    This panel contains sample sizes of primary antibodies against the following housekeeping targets: alpha Tubulin, GAPDH, COXIV, Histone H3, and NaK ATPase.

    The Rabbit Loading Control Antibody Panel is designed for validation and confirmation of western blot analysis when tested in conjunction with your proteins of interest.

Properties

Images

  • All lanes : Anti-GAPDH antibody [EPR16891] (ab181602) at 1/50000 dilution

    Lane 1 : HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysates
    Lane 2 : Xenopus(X. tropicalis) muscle lysates
    Lane 3 : UMNSAH/DF-1 (Transformed chicken embyronic fibroblast cells) whole cell lysates
    Lane 4 : Jurkat (Human T cell leukemia cells from peripheral blood) whole cell lysates

    Lysates/proteins at 20 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

    Predicted band size : 36 kDa
    Observed band size : 36 kDa

    Blocking/Dilution buffer: 5% NFDM/TBST.

  • All lanes : Anti-alpha Tubulin antibody [EPR13478(B)] - Loading Control (ab176560) at 1/1000 dilution (Unpurified)

    Lane 1 : HeLa cell lysate
    Lane 2 : Jurkat cell lysate
    Lane 3 : A431 cell lysate
    Lane 4 : K562 cell lysate

    Lysates/proteins at 10 µg per lane.

    Developed using the ECL technique

    Predicted band size : 50 kDa

    Secondary antibodies - anti-rabbit HRP (ab6721)

  • All lanes : Anti-COX IV antibody [EPR9442(ABC)] - Mitochondrial Loading Control (ab202554) at 1/10000 dilution

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

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

    Predicted band size : 20 kDa
    Observed band size : 17 kDa (why is the actual band size different from the predicted?)


    Exposure time : 3 minutes

    Blocking/Dilution buffer: 5% NFDM/TBST.

  • All lanes : Anti-Histone H3 antibody [EPR16987] - Nuclear Loading Control and ChIP Grade (ab176842) at 1 µg

    Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 10 µg
    Lane 2 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate at 10 µg
    Lane 3 : Drosophila embryo nuclear extract (from melanogaster embryos 0-12hr) at 10 µg
    Lane 4 : S.cerevisiae (Y190) Whole Cell Lysate at 20 µg
    Lane 5 : S.pombe Whole Cell Lysate at 20 µg

    Secondary
    Peroxidase AffiniPure Goat Anti-Rabbit IgG (H+L) at 1/50000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 15 kDa
    Observed band size : 17 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 MES buffer system. The gel was run at 200V for 35 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 ab176842 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution ab133406.

  • All lanes : Anti-Sodium Potassium ATPase antibody [EP1845Y] - Plasma Membrane Loading Control (ab76020) at 1/100000 dilution

    Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates
    Lane 2 : MCF-7 (Human breast adenocarcinoma epithelial cell) whole cell lysates
    Lane 3 : 293 (Human embryonic kidney epithelial cell) whole cell lysates
    Lane 4 : A431 (Human epidermoid carcinoma epithelial cell) whole cell lysates

    Lysates/proteins at 20 µg per lane.

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

    Predicted band size : 113 kDa
    Observed band size : 100 kDa (why is the actual band size different from the predicted?)


    Exposure time : 2 minutes

    Blocking and diluting buffer: 5% NFDM/TBST.

References

This product has been referenced in:
  • Lojewski X  et al. Human iPSC models of neuronal ceroid lipofuscinosis capture distinct effects of TPP1 and CLN3 mutations on the endocytic pathway. Hum Mol Genet 23:2005-22 (2014). Read more (PubMed: 24271013) »
  • Gurbuz I  et al. SAP domain-dependent Mkl1 signaling stimulates proliferation and cell migration by induction of a distinct gene set indicative of poor prognosis in breast cancer patients. Mol Cancer 13:22 (2014). Read more (PubMed: 24495796) »

See all 384 Publications for this product

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

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