产品概述 Ab199713 is a sampler pack of loading control mouse antibodies and a Goat Anti-Mouse IgG secondary antibody conjugated to HRP.This panel contains sample sizes of all mouse host primary antibodies against the following housekeeping targets: alpha Tubulin, GAPDH, COXIV, Histone H3, alpha 1 NaK ATPase.The Mouse Loading Control Antibody Panel is designed for validation and confirmation of western blot analysis when tested in conjunction with your proteins of interest.Please refer to each individual component datasheetfor storage instructions. Explore our range of antibody sample panels designed to provide you with a variety of trial-size antibodies in a convenient and cost-effective format.Refer to the \'Associated Products\' section below to see HRP-conjugated versions of these antibodies. All lanes : Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) at 1 µg/mlLane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell LysateLane 2 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell LysateLane 3 : PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell LysateLysates/proteins at 10 µg per lane.SecondaryAll lanes : Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040)Observed band size: 50 kDa why is the actual band size different from the predicted?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 ab7291 overnight at 4°C. Antibody binding was detected using an anti-mouse antibody conjugated to HRP, and visualised using ECL development solution ab133406 Lanes 1-5 : Anti-GAPDH antibody [mAbcam 9484] - Loading Control (ab9484) ((Blocked in 5% MILK))Lanes 6-10 : Anti-GAPDH antibody [mAbcam 9484] - Loading Control (ab9484)Lanes 1 & 6 : HeLa (Human epithelial carcinoma cell line) Nuclear LysateLanes 2 & 7 : HeLa (Human epithelial carcinoma cell line) Whole Cell LysateLanes 3 & 8 : A431 (Human epithelial carcinoma cell line) Whole Cell Lysate (ab7909)Lanes 4 & 9 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate (ab7899)Lanes 5 & 10 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate (ab7902)Lysates/proteins at 20 µg per lane.SecondaryAll lanes : Goat anti-Mouse (HRP conjugated) at 1/5000 dilutionObserved band size: 40 kDa why is the actual band size different from the predicted?The membrane 1-5 was blocked in 5% Milk (1 hour). The membrane 6-10 was blocked in 5% BSA (1 hour). Abcam routinely uses 5% BSA to block however following recent customer feedback our labs investigated the effect of 5% milk blocking. We can now confirm that milk is not a suitable blocking agent and significantly decreases the signal on the membrane. We have informed all users of the antibody of this finding. All lanes : Anti-COX IV antibody [mAbcam33985] - Mitochondrial Marker (ab33985) at 1 µg/mlLane 1 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell LysateLane 2 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell LysateLane 3 : Skeletal Muscle (Human) Tissue Lysate - adult normal tissue (ab29330)Lane 4 : Skeletal Muscle (Mouse) Tissue LysateLane 5 : Kidney (Cow) Tissue Lysate (ab29073)Lysates/proteins at 10 µg per lane.SecondaryAll lanes : Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilutionObserved band size: 15 kDa why is the actual band size different from the predicted? All lanes : Anti-Histone H3 antibody [mAbcam 24834] - Nuclear Loading Control and ChIP Grade (ab24834) at 1 µg/mlLane 1 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell LysateLane 2 : Testis (Mouse) Tissue LysateLane 3 : PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell LysateLane 4 : Testis (Rat) Tissue Lysate - normal tissue (ab29388)Lysates/proteins at 10 µg per lane.SecondaryAll lanes : Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilutionObserved band size: 18 kDa why is the actual band size different from the predicted? Lanes 1-3 : Anti-alpha 1 Sodium Potassium ATPase antibody [464.6] (ab7671) at 1 µg/mlLanes 4-6 : Anti-alpha 1 Sodium Potassium ATPase antibody [464.6] (ab7671) at 2 µg/mlLanes 7-9 : Anti-alpha 1 Sodium Potassium ATPase antibody [464.6] (ab7671) at 5 µg/mlLanes 1 & 7 : Brain (Human) Tissue Lysate - adult normal tissue (ab29466)Lanes 2 & 5 & 8 : Brain (Mouse) Tissue Lysate (ab27253)Lanes 3 & 6 & 9 : Brain (Rat) Tissue Lysate (ab7942)Lane 4 : Brain (Human) Tissue Lysate - adult normal tissue (ab29466)Lysates/proteins at 20 µg per lane.Observed band size: 112 kDa why is the actual band size different from the predicted? To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on. Click here to view the general protocols 发表研究结果有使用 ab199713？请让我们知道，以便我们可以引用本数据表中的参考文章。 ab199713 被引用在 183 文献中. Gabandé-Rodríguez E et al. High sphingomyelin levels induce lysosomal damage and autophagy dysfunction in Niemann Pick disease type A. Cell Death Differ N/A:N/A (2014).PubMed: 24488099 Beaudry JL et al. Effects of selective and non-selective glucocorticoid receptor II antagonists on rapid-onset diabetes in young rats. PLoS One 9:e91248 (2014).PubMed: 24642683 Konitsiotis AD et al. Attenuation of hedgehog acyltransferase-catalyzed sonic Hedgehog palmitoylation causes reduced signaling, proliferation and invasiveness of human carcinoma cells. PLoS One 9:e89899 (2014).PubMed: 24608521 Uzer G et al. Gap junctional communication in osteocytes is amplified by low intensity vibrations in vitro. PLoS One 9:e90840 (2014).PubMed: 24614887 Arroyo AI et al. Pharmacological reversion of sphingomyelin-induced dendritic spine anomalies in a Niemann Pick disease type A mouse model. EMBO Mol Med 6:398-413 (2014).PubMed: 24448491 View all Publications for this product This is a really good control antibody panel for westernblot. Due to the different antibodies, a large area of the ladder is covered and well separated from the proteins of interest. We used the antibodies at the 1μg / ml concentration and received strong and clear signals.Please note: All products are FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES For licensing inquiries, please contact partnerships@abcam.com