Custom solutions & partnerships Custom antibody development and commercial partnerships to advance your diagnostic and therapeutic discovery. Keep up to date with the latest events Full event breakdown with abstracts, speakers, registration and more View global event calendar Specificity The antibody used for conjugation reacts with mouse immunoglobulins of all classes.Cross-reactions as determined by ELISA for the unconjugated antibody (ab182017): Chicken IgY, less than 2%. Human IgG, less than 6%. Rabbit IgG, less than 7%. Rat IgG, less than 47%. Storage instructions Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle. Store In the Dark. Storage buffer pH: 7.40Preservative: 0.1% Proclin 300 SolutionConstituents: PBS, 1% BSA, 30% Glycerol (glycerin, glycerine) Purification notes This antibody was isolated by affinity chromatography using antigen coupled to agarose beads and conjugated to Horse Radish Peroxidase (HRP). The Abpromise guarantee Our Abpromise guarantee covers the use of ab205719 in the following tested applications. The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user. IP Use at an assay dependent concentration. ELISA Use at an assay dependent concentration. IHC-P 1/2000 - 1/20000. Notes WB1/2000 - 1/20000. IPUse at an assay dependent concentration. ELISAUse at an assay dependent concentration. IHC-P1/2000 - 1/20000. Images Western blot - Goat Anti-Mouse IgG H&L (HRP) (ab205719) All lanes : Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) at 1 µg/mlLane 1 : Liver (Human) Tissue LysateLane 2 : Liver (Mouse) Tissue LysateLane 3 : Liver (Rat) Tissue LysateLane 4 : HeLa (Human epithelial carcinoma cell line) Whole Cell LysateLane 5 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell LysateLane 6 : PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell LysateLysates/proteins at 10 µg per lane.SecondaryAll lanes : Goat Anti-Mouse IgG H&L (HRP) (ab205719) at 1/5000 dilutionDeveloped using the ECL technique.Performed under reducing conditions.Observed band size: 52 kDa why is the actual band size different from the predicted?Exposure time: 5 secondsThis 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 ab205719, and visualised using ECL development solution ab133406. IHC image of alpha tubulin staining in a section of formalin-fixed paraffin-embedded normal human colon tissue*. The section was pre-treated using pressure cooker heat mediated antigen retrieval with sodium citrate buffer (pH6) for 30mins, and incubated overnight at +4 C with ab7291 at 1/1000 dilution. An HRP-conjugated secondary (Ab205719, 1/10000 dilution) was used to detect the primary for 1hr at room temperature. DAB was used as the chromogen (ab103723), diluted 1/100 and incubated for 10min at room temperature. The section was counterstained with haematoxylin and mounted with DPX. The inset negative control image is taken from an identical assay without primary antibody.For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre All lanes : Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) at 1 µg/mlLane 1 : Liver (Human) Tissue LysateLane 2 : Liver (Mouse) Tissue LysateLane 3 : Liver (Rat) Tissue LysateLysates/proteins at 10 µg per lane.SecondaryAll lanes : ab205719 (Left Image) at 1/5000 and a competitor secondary (Right Image) at 1/5000. Notice the decreased signal of the competitor product.Performed under reducing conditions.Observed band size: 52 kDa why is the actual band size different from the predicted?Exposure time: 5 secondsThis 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 ab205719 (Left Image) and a competitor secondary (Right Image), and visualised using ECL development solution ab133406. IHC image of histone H4 staining in a section of formalin-fixed paraffin-embedded normal human colon tissue*. The section was pre-treated using pressure cooker heat mediated antigen retrieval with sodium citrate buffer (pH6) for 30mins, and incubated overnight at +4 C with ab31830 at 1/1000 dilution. An HRP-conjugated secondary (Ab205719, 1/10000 dilution) was used to detect the primary for 1hr at room temperature. DAB was used as the chromogen (ab103723), diluted 1/100 and incubated for 10min at room temperature. The section was counterstained with haematoxylin and mounted with DPX. The inset negative control image is taken from an identical assay without primary antibody.For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre All lanes : No Primary AntibodyLane 1 : Liver (Human) Tissue LysateLane 2 : Liver (Mouse) Tissue LysateLane 3 : Liver (Rat) Tissue LysateLysates/proteins at 10 µg per lane.SecondaryAll lanes : ab205719 (Left Image) 1/2000 and a competitor secondary (Right Image) 1/2000. Notice the increased background of the competitor product.Performed under reducing conditions.Exposure time: 10 secondsThis 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 incubated overnight with 2% Bovine Serum Albumin at 4 C. Any non-specific background binding was assessed by incubating the membrane with ab205719 (Left Image) and a competitor secondary (Right Image), and visualised using ECL development solution ab133406. All lanes : Anti-beta Actin antibody [mAbcam 8226] - Loading Control (ab8226) at 1 µg/mlLane 1 : Liver (Human) Tissue LysateLane 2 : Liver (Mouse) Tissue LysateLane 3 : Liver (Rat) Tissue LysateLane 4 : HeLa (Human epithelial carcinoma cell line) Whole Cell LysateLane 5 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell LysateLane 6 : PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell LysateLysates/proteins at 10 µg per lane.SecondaryAll lanes : Goat Anti-Mouse IgG H&L (HRP) (ab205719) at 1/5000 dilutionDeveloped 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: 10 secondsThis 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 using2% Bovine Serum Albumin before being incubated with ab8226 overnight at 4 C. Antibody binding was detected using ab205719, and visualised using ECL development solution ab133406. Cross-reactivity of the polyclonal secondary antibody ab182017 was tested using a sandwich ELISA approach. The wells were coated with the indicated IgG standards at 1 g/ml (50 l/well) and incubatedovernight at 4 C, followed by a 5% BSA blocking step for 2h at RT. ab182017 was then added starting at 1 g/ml and gradually diluted 1/4 (50 l/well), followed by incubationfor 2h. For the detection Donkey anti-Goat IgG H L (HRP) (ab6885) was used at 1/10,000 dilution (50 l/well), followed by incubationfor 1h at RT.Fot the batch tested, ab182017 showed a cross-reactivity below 2% towards Chicken IgY, 6% towards Human IgG, 7% towards Rabbit IgG and 47% towards Rat IgG.This data was developed using the unconjugated antibody (ab182017). Cross-reactivity of Goat anti-Mouse IgG H L (ab182017)and Goat anti-Mouse IgG H Lobtained from two different vendors was tested using a sandwich ELISA approach. The wells were coated with the indicated IgG standards (Rabbit, Human, Mouse and Rat) at 1 g/ml(50 l/well) and incubatedovernight at 4 C, followed by a 5% BSA blocking step for 2h at RT. Secondary antibodies were then added starting at 1 g/ml and gradually diluted 1/4 (50 l/well), followed by incubationfor 2h. For the detection Donkey anti-Goat IgG H L (HRP) (ab6885)was used at 1/10,000 dilution (50 l/well), followed by incubationfor 1h atRT. This data is from a representative dilution.This data was developed using the unconjugated antibody (ab182017). 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 Publishing research using ab205719? Please let us know so that we can cite the reference in this datasheet. ab205719 has been referenced in 397 publications. Yu C et al. CircRNA TGFBR2/MiR-25-3p/TWIST1 axis regulates osteoblast differentiation of human aortic valve interstitial cells. J Bone Miner Metab 39:360-371 (2021).PubMed: 33070258 Liu BQ et al. Fibrinogen-like protein 2 promotes the accumulation of myeloid-derived suppressor cells in the hepatocellular carcinoma tumor microenvironment. Oncol Lett 21:47 (2021).PubMed: 33281958 Peng T et al. miR-125/CDK2 axis in cochlear progenitor cell proliferation. Mol Med Rep 23:N/A (2021).PubMed: 33300064 Wang X et al. Penehyclidine hydrochloride alleviates lipopolysaccharide-induced acute respiratory distress syndrome in cells via regulating autophagy-related pathway. Mol Med Rep 23:N/A (2021).PubMed: 33300058 Zhao FY et al. PDGF mediates pulmonary arterial smooth muscle cell proliferation and migration by regulating NFATc2. Mol Med Rep 23:N/A (2021).PubMed: 33179105 View all Publications for this product - 20ug of bovine liver lysates were loaded on to 4-15% gradient gel- Blocked for an hour in 5% skim milk- 1:4000 primary antibody used (ab110413). Incubation O/N @4C- 1:20000 secondary antibody used (ab205719). Incubation 1 Hour room temperature. Whole Cell lysate - human ES cells.Loading amount: 10 µg.Gel Running Conditions: Reduced Denaturing (10%).Blocking step: 5%BSA/1%OvaAlb/PBS as blocking agent for 3 hours at room temperature.Primary antibody: Nanog diluted 1:1000 in 1%BSA/0.2%OvaAlb/PBS 13hours at 4°C.Secondary antibody: ab205719 - Goat Anti-Mouse IgG H&L (HRP) diluted 1:5000 in TBST.ECL prime: 4 second exposure.Predicted size: ~45kDa. Excellent specificity.Excellent sensitivity.WB conditions:- Cell lisates: 20ug protein- non-reducing- Primary antibody: Mouse anti-GRP78, clon 40/BiP, Cat. No. 610979. BD Biosciences. Dilution 1:1000- Secondary antibody: 1:6000- Developer: ECL Prime, Amersham, GE Helathcare Life Sciences, Product code RPN2232- Exposure time: 1 second, 3th film out 4Please note: All products are FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES For licensing inquiries, please contact partnerships@abcam.com