Brd4抗体(ab75898)| Abcam中文官网

来自 : 发布时间：2024-05-09

长期批量供应 —— 采用重组技术，可实现快速生产 首次实验即可成功 —— 经过大量验证确认了特异性 符合伦理标准 —— 产品不含动物成分 Synthetic peptide corresponding to Human BRD4 aa 1100-1200 conjugated to keyhole limpet haemocyanin.(Peptide available as ab85811) 常规说明 \"Note: Batches of this product that have a concentration The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q As 存放说明 Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle. 存储溶液 pH: 7.40Preservative: 0.02% Sodium azideConstituent: PBSBatches of this product that have a concentration The Abpromise guarantee Abpromise™承诺保证使用ab75898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。 Use a concentration of 1 µg/ml. Detects a band of approximately 152 kDa (predicted molecular weight: 152 kDa). WBUse a concentration of 1 µg/ml. Detects a band of approximately 152 kDa (predicted molecular weight: 152 kDa). 疾病相关 Note=A chromosomal aberration involving BRD4 is found in a rare, aggressive, and lethal carcinoma arising in midline organs of young people. Translocation t(15;19)(q14;p13) with NUT which produces a BRD4-NUT fusion protein. All lanes : Anti-Brd4 antibody (ab75898) at 1 µg/mlLane 1 : Wild-type HAP1 cell lysateLane 2 : BRD4 knockout HAP1 cell lysateLysates/proteins at 20 µg per lane.Performed under reducing conditions.Predicted band size: 152 kDaObserved band size: 220 kDa why is the actual band size different from the predicted? Lanes 1 - 2: Merged signal (red and green). Green - ab75898 observed at 220 kDa. Red - loading control Mouse anti Vinculin observed at 125 kDa.ab75898 was shown to react with Brd4 in HAP1 wild-type cells in Western blot with loss of signal observed in BRD4 knockout sample. Wild-type HAP1 and BRD4 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween ) before incubation with ab75898 and Mouse anti Vinculin overnight at 4 C at 1 g/ml and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H L (IRDye 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H L (IRDye 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging. All lanes : Anti-Brd4 antibody (ab75898) at 1 µg/mlLane 1 : Liver (Mouse) Tissue Lysate Lane 2 : Liver (Rat) Tissue Lysate Lane 3 : Human liver tissue lysate - total protein (ab29889)Lysates/proteins at 10 µg/ml per lane.SecondaryAll lanes : Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) Developed using the ECL technique.Performed under reducing conditions.Predicted band size: 152 kDaObserved band size: 171 kDa why is the actual band size different from the predicted?Additional bands at: 70 kDa. We are unsure as to the identity of these extra bands.Exposure time: 150 seconds Immunocytochemistry/ Immunofluorescence - Anti-Brd4 antibody (ab75898)Image courtesy of an Abreview submitted by Dr. Kirk McManus, Univ. of Manitoba/Cancer Care MICB, Canada ab75898 (1/500) staining Brd4 in HeLa cells (green). Cells were fixed in paraformaldehyde, permeabilized with 0.5% Triton X-100/PBS and counterstained with DAPI in order to highlight the nucleus (red). For further experimental details please refer to Abreview.See Abreview ICC/IF image of ab75898 stained MCF7 cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab75898, 5µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. This antibody also gave a positive result in 4% PFA fixed (10 min) HeLa, Hek293 and HepG2 cells at 5µg/ml. 发表研究结果有使用 ab75898？请让我们知道，以便我们可以引用本数据表中的参考文章。 ab75898 被引用在 22 文献中. Sui S et al. Ferritinophagy is required for the induction of ferroptosis by the bromodomain protein BRD4 inhibitor (+)-JQ1 in cancer cells. Cell Death Dis 10:331 (2019).PubMed: 30988278 Pan M et al. miR-125b-mediated regulation of cell proliferation through the Jagged-1/Notch signaling pathway by inhibiting BRD4 expression in psoriasis. Mol Med Rep 19:5227-5236 (2019).PubMed: 31059052 Wang C et al. miR-204 enhances p27 mRNA stability by targeting Brd4 in head and neck squamous cell carcinoma. Oncol Lett 16:4179-4184 (2018).PubMed: 30250532 Chakraborty D et al. A BET Bromodomain Inhibitor Suppresses Adiposity-Associated Malignant Transformation. Cancer Prev Res (Phila) 11:129-142 (2018).PubMed: 29246955 Wang Y et al. Bromodomain-containing protein 4 is critical for the antiproliferative and pro-apoptotic effects of gambogic acid in anaplastic thyroid cancer. Int J Mol Med 42:161-170 (2018).PubMed: 29717765 View all Publications for this product Type Cross-linking (X-ChIP)Duration of cross-linking step: 10 minute(s) and 0 second(s)Specification of the cross-linking agent: Formaldehyde Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) Concentration: 3% Temperature: 25 C Thank you for taking time to complete our questionnaire. I am sorry to hear that this antibody is not providing satisfactory results. The details provided will enable us to investigate this case and will provide us with vital information for monitoring product quality.Having reviewed this case, I would like to offer some suggestions to help optimize the results from ab75898. When testing our antibodies, our lab uses 5% BSA as a blocking reagent, so I recommend switching to this instead of milk. For unknown reasons some antibodies give stronger, more specific signals on blots blocked with BSA instead of milk, so doing this may improve the results you are seeing, and reduce the non-specific bands. An example of the above is the western blot obtained with the Abcam GAPDH antibody ab9385 : https://www.abcam.com/gapdh-antibody-hrp-loading-control-ab9385.html#GAPDH-Primary-antibodies-ab9385-3.jpg (or use the following: https://www.abcam.com/ab9385).BSA should then also be used in the dilutionbuffer of the antibodies instead of milk. I would also suggest reducing the level of blocking agent in the antibody diluents to 0.1% initially to see if you can observe a signal. In order to prevent the over washing of the membrane I would also suggest reducing the wash steps to 3x5 minutes after each incubation.Should the suggestions not improve the results, please do let me know.In the event that a product is not functioning in the species and applications cited on the product datasheet (and the problem has been reported within 6 months of purchase), we would be pleased to provide a free of charge replacement, credit note, or refund.I hope this information is helpful, and I thank you for your cooperation. Read MoreI am very sorry to hear that you are having problem with this antibody. As I understand from your e-mail, there is some problem with co-IP. Unfortunately, as the datasheet indicates this product has been tested in ICC/IF, WB applications only - but not in IP yet. This means that we do not have any IP data to share with our customers.Nevertheless, if you wish me to look through the details of the protocol, I could certainly do it for you in case some small alterations could be implemented to improve the signal. Please could you complete the questionnaire attached to this e-mail. Regarding the epitope sequence - ab75898: anti-Brd4 antibody is a polyclonal antibody. Polyclonal antibodies consist of a mixed population of IgGs each of which will recognize a different epitope. I hope this information is helpful to you. Please do not hesitate to contact us if you need any additional information or advice. Read MorePlease note: All products are FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES For licensing inquiries, please contact partnerships@abcam.com