存放说明 Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle. Please see notes section. Store In the Dark. 存储溶液 Preservative: 0.01% Gentamicin sulphateConstituents: 0.42% Potassium phosphate, 0.88% Sodium chloride 纯化说明 This product was prepared from monospecific antiserum by immunoaffinity chromatography using Rabbit IgG coupled to agarose beads. 常规说明 HRP conjugated anti-rabbit secondary antibody optimized for western blot and immunohistochemistry. Some customers reported seeing brown precipitates in the vials. The brown precipitates are very common with HRP conjugated antibodies; we suggest vortexing the vial and using this antibody as normal. Our customer s feedback says the antibody worked great. If in case the antibody fails to give results then please contact our Scientific Support team for assistance.Centrifuge product if not completely clear after standing at room temperature. Dilute only prior to immediate use. The Abpromise guarantee Abpromise™承诺保证使用ab6721于以下的经测试应用 \"应用说明”部分 下显示的仅为推荐的起始稀释度；实际最佳的稀释度/浓度应由使用者检定。 1/2000 - 1/20000. Suggested working dilution of 1/3000 (see PMID: 17222046). In addition, found to work at 1/20000 (see PMID: 16936283).Working dilutions are highlighted in the table below. Please note that the antibody can be diluted to 1:48,000 to 1:207,000 in many instances. ELISA 1/120000. Immunomicroscopy Use at an assay dependent concentration. Dot blot Use at an assay dependent concentration. ICC 1/1000 - 1/5000. IHC-Fr 1/1000. 说明 IHC-P1/1000. WB1/2000 - 1/20000. Suggested working dilution of 1/3000 (see PMID: 17222046). In addition, found to work at 1/20000 (see PMID: 16936283).Working dilutions are highlighted in the table below. Please note that the antibody can be diluted to 1:48,000 to 1:207,000 in many instances. ELISA1/120000. ImmunomicroscopyUse at an assay dependent concentration. Dot blotUse at an assay dependent concentration. ICC1/1000 - 1/5000. IHC-Fr1/1000. 图片 Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Goat Anti-Rabbit IgG H&L (HRP) (ab6721)Farah et al PLoS One. 2018 Feb 2;13(2):e0191526. doi: 10.1371/journal.pone.0191526. eCollection 2018. Fig 3. Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/ Anti-phospho-tau immunostaining in Dryocopus lineatus.Tau-positivity in the midbrain (Panel A (shown)and B) and the corpus callosum (C) of the Dryocopus lineatus brain. The axonal tract staining demonstrates a thread-like pattern, similar to that seen with Gallyas sliver staining. Occasional intracellular tau-accumulations were identified within neurons (D).For full method, please see paper. Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab6721)This image is courtesy of an Abreview submitted by Maryna Polyakova. All lanes : Anti-NEK7 antibody (ab80948) at 1/2000 dilutionAll lanes : Mouse brain tissue cytoplasmic lysateLysates/proteins at 10 µg per lane.SecondaryAll lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab6721) at 1/5000 dilutionDeveloped using the ECL technique.Performed under reducing conditions.Exposure time: 10 secondsBlocked with 5% non-fat milk for 1 hour at 18 C Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Goat Anti-Rabbit IgG H&L (HRP) (ab6721)This image is courtesy of an anonymous Abreview. ab3580 staining glucocorticoid receptor in mouse epididymis tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections).Tissue was fixed with Bouin\'s solution and blocked with 1.5% serum for 30 minutes at 25 antigen retrieval was by heat mediation in a citrate buffer. Samples were incubated with primary antibody (1/1000) for 14 hours at 4 C. An HRP-conjugated goat anti-rabbit IgG H L (ab6721) (1/200) was used as the secondary antibody.See Abreview Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Goat Anti-Rabbit IgG H&L (HRP) (ab6721)Image is courtesy of an anonymous AbReview. Immunohistochemical analysis of PFA-fixed paraffin-embedded rat cardiac tissue sections, labeling Conexin 43 with ab117843 at a dilution of 1/500 incubated for 12 hours at 4 C in 1% BSA in TBS. Antigen retrival was via Tris-EDTA pH 9.0 (heat mediated). Blocking was 3% BSA incubated for 1 hour at 37 C. The secondary was ab6721 at 1/500.See Abreview ab6721 was used at dilution 1/100 with the primary antibody ab11370 in IHC-P. See the review on ab11370. ab6721 was used at dilution 1/100 with the primary antibody ab35604 in IHC-Fr. See the review on ab35604. 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 ab6721？请让我们知道，以便我们可以引用本数据表中的参考文章。 ab6721 被引用在 2839 文献中. Tian Y et al. LncRNA LINC00974 Downregulates miR-122 to Upregulate RhoA in Oral Squamous Cell Carcinoma. Cancer Biother Radiopharm 36:18-22 (2021).PubMed: 31702382 Zhang H et al. SDF-1 mediates mesenchymal stem cell recruitment and migration via the SDF-1/CXCR4 axis in bone defect. J Bone Miner Metab 39:126-138 (2021).PubMed: 33079278 Min J et al. Phenotype and biological characteristics of endometrial mesenchymal stem/stromal cells: A comparison between intrauterine adhesion patients and healthy women. Am J Reprod Immunol 85:e13379 (2021).PubMed: 33206449 Song T et al. Zearalenone exposure affects the Wnt/ß-catenin signaling pathway and related genes of porcine endometrial epithelial cells in vitro. Anim Biosci 34:993-1005 (2021).PubMed: 32898953 Yu L et al. Long non-coding RNA HAND2-AS1 inhibits gastric cancer progression by suppressing TCEAL7 expression via targeting miR-769-5p. Dig Liver Dis 53:238-244 (2021).PubMed: 32952069 View all Publications for this product I have been using this secondary antibody for my western blotting experiments and its working fine with all of proteins of our interest (i.e. Rodent CYP-450 enzymes like CYP-3A, 2D1, 2E1, 1A2, 2B1 etc.)Before using this I had also explored the HRP antibody from other vendors but they were not so specific I was having lot of back ground noise on my membranes irrespective of varying dilutions and incubation times and so on. So I would highly recommend using this for western blot experiments. The dilution used for this secondary antibody was 1/20000 and we obtained intesnse specific bands even at very short incubation times. It works very well with phosphorylated proteins which sometimes are harder to detect (in these cases we use 1/10000 dilution or 1/5000). Very clear signal. The image is of a cross section of 3 metatarsal bones, primary antibody is Periostin (ab14041). Secondary antibody (ab6721) was used at a dilution of 1:5000 in 5% BSA. Placental lysate was used. primary antibody was purchased for VEGFR1 (ab 32152) antibody. gel: 10%, 200 V for 35 min.transfer 17 V for 35 min. using Biorad TURBO Blocking 5% BSA. Experimental conditions:Bone tissue lysate (tibia) extracted with RIPA buffer10% SDS-PAGEBlocking 1h with 5% milk in TBS-T at RTIncubation with primary AB 1:1000 overnight at 4ºCIncubation with secondary AB (ab6721 - Goat Anti-Rabbit IgG) 1:2000 for 1h at RTDeveloped with ECL for 537sResultsNo nonspecific binding. No background. Clear bands.In the case of ab6721, the storage condition has changed because we removed the glycerol bit from the antibody storage buffer. As a consequence, from now on we recommend to store the antibody at 4 degrees. For more information on antibody storage and stability, please visit our Antibody Storage Guide on our Protocols and Troubleshooting Tips page (https://www.abcam.com/index.html?pageconfig=popular_protocols). Read More The dilution factor on the datasheet only a recommendation for starting. The actual dilutions need to be optimized by the scientist since it depends on many factors such expression levels of the target in the samples, primary antibody (clonality, purification, specificity), signal amplification, detection system etc. Read MorePlease note: All products are FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES For licensing inquiries, please contact partnerships@abcam.com