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 Product overview DAB substrate kit ab64238 is used for IHC staining using HRP / peroxidase enzyme for detection. Reagent is stable for up to 6 hours after mixing the two components.DAB (3,3\'Diaminobenzidine) is the most commonly used chromogen for immunohistochemical staining. In the presence of HRP / peroxidase, DAB produces a brown precipitate that is insoluble in alcohol.This product is a two component form consisting of a liquid, refrigerator stable DAB Chromogen and DAB Substrate.IHC protocol suitable for use with DAB substrate kit:For frozen sections, skip steps 1 and 2.1. Deparaffinize and rehydrate formalin-fixed paraffin-embedded tissue section.2. Use appropriateantigen retrieval buffer or enzyme(primary antibody dependent) to treat sections. Wash 3 times in buffer.3. Add enoughhydrogen peroxide blocking solutionto cover the sections. Incubate for 10 minutes. Wash 2 times in buffer.If necessary, useavidin biotin blocking.4. Applyprotein block(or normal serumfrom same species as secondary antibody) and incubate for 5 minutes at room temperature to block nonspecific background staining. Wash once in buffer.5. Apply primary antibody inantibody diluentand incubate.6. Wash 4 times in buffer. Incubate slide withbiotinylated secondary antibody(or HRP polymer secondary antibody and skip step 7). Wash 4 times in buffer.7.Apply streptavidin-HRP and incubate for 10 minutes at room temperature.8. Rinse 4 times in buffer. Place slide in DAB substrate ab64238 orAEC substrateand incubate until desired color is achieved (1-10 mins). Rinse 4 times in buffer.9. Add enough drops of hematoxylinto cover the section. Incubate for 1 minute.10. Rinse 7-8 times in tap water. Addmounting mediumto cover the section. Notes Find complete IHC kits, and reagents for antigen retrieval, blocking, signal amplification, visualization, counterstaining, and mounting in theIHC kits and reagents guide. Storage buffer Constituents: 100% Propylene glycol, 5% Hydrogen peroxide, DAB substrate buffer Relevance 3,3\' Diaminobenzidine (DAB) is a widely used chromogen for immunohistochemical staining. In the presence of peroxidase enzyme, DAB produces a brown precipitate that is insoluble in alcohol. The Abpromise guarantee Our Abpromise guarantee covers the use of ab64238 in the following tested applications. The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user. IHC-P Use at an assay dependent dilution. Notes IHC-PUse at an assay dependent dilution. Images Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)Lean QY et al. PLoS One (2015) Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/ Immunohistochemical analysis of healthy and colitis mouse colon sections (untreated and treated with enoxaparin) labeling claudin-4 with ab15104 at 1/200. ab7090, anti-rabbit immunoglobulin G conjugated to horseradish peroxidase (HRP) at 1/300 was used as the secondary antibody. ab64238 at 1/50 was used to develop the histological signal.Antigen retrieval was performed by incubating the sections for 10 minutes at 97 C in 1 mM EDTA buffer, pH 8.0 or 10 mM citrate buffer, pH 6.0.Control, C; untreated colitis, DSS; oral enoxaparin, OE; intraperitoneal injection of enoxaparin, IPE. Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)Kumar P et al. PLoS One (2014) Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/ Immuno-staining offormalin fixed and paraffin-embedded liver sections with ab64238. Secontions were obtained from control (saline) and treated with CCl4+CT-N and CCl4+AD-N (left) and antibody control (right). Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)Jujo T et al. PLoS One (2014) Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/ Immunohistochemical analysis of mouse lung. Sampleswere fixed in 10% buffered formalin, paraffinized and sliced at 1.5 m thick. Antigen retrieval was performed usingab64214 for the deparaffinized slices. Sections were blocked with 2% normal goat serum, PBS(-) and 0.1% Tween20.They were then incubated with the primary antibodies for 1 hour at 4 C and with secondary antibodies for 30 minutes at room temperature. The avidin-biotin-peroxidase complex method with peroxidase streptavidin and the DAB substrate kit ab64238 was performed.(A) A resected lung from a mouse sacrificed 28 days after SCL injection.(B) Haemotoxylin and Eosin staining showing the tumor composed of a central area with necrosis and a peripheral zone filled with SCLs.(C) Immunohistochemical staining for MMP-14 showing positive expression of MMP-14 in the peripheral zone of the tumor and negative in central zone.(D) Immunohistochemical staining weak staining of MMP-2 in the peripheral zone. Publishing research using ab64238? Please let us know so that we can cite the reference in this datasheet. ab64238 has been referenced in 103 publications. Gualandi M et al. Oncogenic ALKF1174L drives tumorigenesis in cutaneous squamous cell carcinoma. Life Sci Alliance 3:N/A (2020).PubMed: 32312912 Albarracin L et al. Efficient Selection of New Immunobiotic Strains With Antiviral Effects in Local and Distal Mucosal Sites by Using Porcine Intestinal Epitheliocytes. Front Immunol 11:543 (2020).PubMed: 32322251 Jiang W et al. Long non-coding RNA EBLN3P promotes the recovery of the function of impaired spiral ganglion neurons by competitively binding to miR-204-5p and regulating TMPRSS3 expression. Int J Mol Med 45:1851-1863 (2020).PubMed: 32186779 Matouskova K et al. Exposure to low doses of oxybenzone during perinatal development alters mammary gland morphology in male and female mice. Reprod Toxicol 92:66-77 (2020).PubMed: 31408669 Samanta J et al. Oleic Acid Protects from Arsenic-Induced Cardiac Hypertrophy via AMPK/FoxO/NFATc3 Pathway. Cardiovasc Toxicol 20:261-280 (2020).PubMed: 31571030 View all Publications for this product Gives good results at a dilution 1:50. Used on brain sections with an antibody against PLP CNPase (ab6319) staining on murine brain tissue. Visualization step using ab64238 DAB substrate (1:50 dilution, 10min incubation time) This DAB substrate kit (ab94665) is easy to use, and the quality is good. Liyong Zhang This kit is streamlined to use at ease. The results is fantastic. Liyong Zhang Thank you for contacting us. The kit will work with the rabbit and mouse primary antibodies. You can try citrate buffer pH for a start and Tris/EDTA in case the staining does not work. Please check the following for protocol and troubleshooting of;https://www.abcam.com/index.html?pageconfig=resource&rid=11384https://www.abcam.com/index.html?pageconfig=resource&rid=11467I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information. Free Rabbit monoclonal antibody with any purchase of a primary antibody, while stocks last! Quote \"RABMAB-XBSMG” in your next primary antibody order. For more information, visit the following link: https://www.abcam.com/index.html?pageconfig=resource&rid=15447 Read More Thank you for contacting us. Heat mediated antigen retrieval method with microwave or pressure cooker is recommended for this ab. The suitable detection system depends on the substrate you want to use. e.g. with DAB or AEC, we have following products; https://www.abcam.com/DAB-Enhancer-ab675.htmlhttps://www.abcam.com/DAB-Substrate-Kit-ab64238.htmlhttps://www.abcam.com/AEC-Substrate-System-Ready-to-Use-ab64252.htmlI hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information. Free Rabbit monoclonal antibody with any purchase of a primary antibody, while stocks last! Quote \"RABMAB-XBSMG” in your next primary antibody order. For more information, visit the following link: https://www.abcam.com/index.html?pageconfig=resource&rid=15447 Read More Thank you for contacting us again. ab1002 Anti-Rabies Virus antibody looks very promising as it has been established for the use in IHC-P and should detect a wide range of rabies strains. As a starting dilution the customer can try a 5 ug/ml dilution incubated overnight at 4C. However, it may be that the actual optimum dilution may be lower or higher, but this depends on her samples. Another point is that I would strongly recommend a heat induced Antigen Retrieval if her slides are fixed with formalin.https://www.abcam.com/Rabies-Virus-antibody-1C5-ab1002.html (or use the following: https://www.abcam.com/Rabies-Virus-antibody-1C5-ab1002.html).A matching secondary antibody would be Goat polyclonal Secondary Antibody to Mouse IgG2a - heavy chain (Biotin), pre-adsorbed (ab98696)https://www.abcam.com/Goat-Mouse-IgG2a-heavy-chain-Biotin-preadsorbed-ab98696.html (or use the following: https://www.abcam.com/Goat-Mouse-IgG2a-heavy-chain-Biotin-preadsorbed-ab98696.html).To be fair, I am not the biggest fan of ready- to- use solution (andmost of my colleagues here a neither), for the following reason: most of all antibodies need optimization, may it be the blocking step, the retrieval time or method, the antibody incubation buffer etc. With ready-mix solutions it almost impossible to adjust the experiment settings. Ready mix solution make sense if the antibody is used in a high -throughput lab (like 20 slides at once with a fixed protocol), but not if you have to learn IHC-P and have the freedom to adjust the protocol fore the perfect results. In addition, it\'s cheaper ;)Of course, once the protocol is optimized, using ready mixed solution are a very time efficient and make life easier, but I wouldn\'t use them as a beginner.All the buffers your customer needs (see attachment) are listed in our very detailed and easy to follow step-by-step our protocol. However, none of my advice is written in stone, so if your customer prefers the ready mix solutions, we are more than happy to sell them to her.Something we cannot provide are the reagents for the de-paraffinization, I am afraid, but a company like Sigma should be able to help with that.Pease sent the customer our protocol and my advice. I am more than happy to assist her to get her experiments running. And another thing: she shouldn\'t be shy to ask any question she may have about the protocol!I hope this more information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.Use our products? Submit an Abreview. Earn rewards! https://www.abcam.com/abreviews Read More Thank you for contacting us.Here is the list with the requested items. However, please be aware that this is not a technical inquiry as all item could be found by using the search application on our website under the reagents category. Please let us know if you need help on navigating our website so you can help your customers faster than relying on us.Regarding the Rabies antibody, I am afraid we do not have what your customer requests: All we have are mouse monoclonal antibodies against this virus, but none produced in goat. Please let me know if this expectable for her, so I can suggest the right primary antibody with the matching biotinylated secondary antibody.https://www.abcam.com/10x-Citrate-Buffer-pH-60-ab64214.html (or use the following: https://www.abcam.com/10x-Citrate-Buffer-pH-60-ab64214.html). https://www.abcam.com/10x-EDTA-Buffer-pH-80-ab64216.html (or use the following: https://www.abcam.com/10x-EDTA-Buffer-pH-80-ab64216.html). https://www.abcam.com/DAB-Substrate-Kit-ab64238.html (or use the following: https://www.abcam.com/DAB-Substrate-Kit-ab64238.html). https://www.abcam.com/Streptavidin-HRP-ready-to-use-ab64269.html (or use the following: https://www.abcam.com/Streptavidin-HRP-ready-to-use-ab64269.html). https://www.abcam.com/Hydrogen-Peroxide-Blocking-Reagent-ab94666.html (or use the following: https://www.abcam.com/Hydrogen-Peroxide-Blocking-Reagent-ab94666.html). https://www.abcam.com/Protein-Block-ab128988.html (or use the following: https://www.abcam.com/Protein-Block-ab128988.html). https://www.abcam.com/Mayers-Hematoxylin-ab128990.html (or use the following: https://www.abcam.com/Mayers-Hematoxylin-ab128990.html). I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.Use our products? Submit an Abreview. Earn rewards! https://www.abcam.com/abreviews Read MorePlease note: All products are FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES For licensing inquiries, please contact partnerships@abcam.com