使用说明：
1. 对于组织切片或细胞样品或膜，在与辣根过氧化物酶标记的抗体或其它形式的探针孵育后，用适当洗涤
   液洗涤3-5次，每次3-5分钟。对于检测内源性辣根过氧化物酶的组织或细胞样品，在适当固定后，也用
   适当洗涤液洗涤3-5次，每次3-5分钟。
2. 按照如下比例依次加入各溶液，混匀后即配制成DAB染色工作液：

3. 最后一次洗涤完毕后，去除洗涤液，加入适量DAB染色工作液，确保能充分覆盖样品。
4. 室温避光孵育3-30分钟或更长时间(可长达24小时)，直至显色至预期深浅。
5. 去除DAB染色工作液，用蒸馏水洗涤1-2次即可终止显色反应。
6. 对于组织切片或细胞样品，显色反应终止后，如有必要可以用中性红染色液(neutral red staining
   solution)染色，以便于观察。对于膜，显色反应终止后，可以室温晾干避光保存。

常见问题：
1. 背景显色太深。
   a. 在免疫组化时如果背景显色太深，一方面需考虑使用适当的封闭液进行封闭，例如选购适当的封闭液
      或使用和一抗相同来源的血清(10%)进行封闭。另一方面，请注意选购经过适当吸附的二抗，以减小
      二抗的非特异性吸附。
   b. 在免疫组化时如果背景显色太深，需注意灭活内源性过氧化氢酶。可以在4体积甲醇中加入1体积3%过
      氧化氢，混匀后用于内源性过氧化氢酶的灭活。
   c. 可以考虑缩短显色时间，或降低二抗浓度。另外，选择适当强度的洗涤液，或延长洗涤时间也会有所
      帮助。
2. 没有显色或显色太弱。
   a. 可以考虑适当提高一抗或二抗的浓度。检测二抗效果，滴一滴稀释二抗在膜上，检测二抗是否可以被
      正常显色。
   b. 可以考虑使用更加灵敏的放大检测体系，例如使用生物素检测体系。
   c. 可以适当延长显色时间，另外确定抗原修复是否对于使用的一抗是必需的。

使用本产品的相关论文：
1. Li L, Qi Z, Qian J, Bi F, Lv J, Xu L, Zhang L, Chen H, Jia R.
   Induction of apoptosis in hepatocellular carcinoma Smmc-7721 cells by vitamin K(2) is
   associatedwith p53 and independent of the intrinsic apoptotic pathway.
   Mol Cell Biochem. 2010 Sep;342(1-2):125-31.
2. Wang ZH, Xu L, Wang Y, Cao MQ, Li L, Bai T.
   Clinicopathologic correlations between human papillomavirus 16 infection and Beclin 1
   expressionin human cervical cancer.
   Int J Gynecol Pathol. 2011 Jul;30(4):400-6.
3. Gao X, Luo Z, Xiang T, Wang K, Li J, Wang P.
   Dihydroartemisinin induces endoplasmic reticulum stress-mediated apoptosis in HepG2
   human hepatoma cells.
   Tumori. 2011 Nov-Dec;97(6):771-80.
4. Cao J, Meng F, Gao X, Dong H, Yao W.
   Expression and purification of a natural N-terminal pre-ligand assembly domain of tumor
   necrosisfactor receptor 1 (TNFR1 PLAD) and preliminary activity determination..
   Protein J. 2011 Apr;30(4):281-9.
5. Ren J, Li W, Yan L, Jiao W, Tian S, Li D, Tang Y, Gu G, Liu H, Xu Z.
   Expression of CIP2A in renal cell carcinomas correlates with tumour invasion, metastasis
   and patients' survival.
   Br J Cancer. 2011 Dec 6;105(12):1905-11.
6. Ren J, Liu H, Yan L, Tian S, Li D, Xu Z.
   Microvessel density and heparanase over-expression in clear cell renal cell cancer:
   correlationsand prognostic significances.
   World J Surg Oncol. 2011 Dec 2;9:158.
7. Li W, Zhao L, Zang W, Liu Z, Chen L, Liu T, Xu D, Jia J.
   Histone demethylase JMJD2B is required for tumor cell proliferation and survival and
   isoverexpressed in gastric cancer.
   Biochem Biophys Res Commun. 2011 Dec 16;416(3-4):372-8.
8. Wang T, You N, Tao K, Wang X, Zhao G, Xia N, Li N, Tang L, Liu W, Dou K.
   Notch is the key factor in the process of fetal liver stem/progenitor cells
   differentiation intohepatocytes.
   Dev Growth Differ. 2012 Jun;54(5):605-17.
9. Li Y, Wu S, Ouyang J, Mao L, Li W, Lin H.
   Expression of insulin-like growth factor-1 of orange-spotted grouper (Epinephelus
   coioides)inyeast Pichia pastoris.
   Protein Expr Purif. 2012 Jul;84(1):80-5.
10.Liu H, Liu G, Gong L, Zhang Y, Jiang G.
   Local suppression of IL-21 in submandibular glands retards the development of
   Sjögren'ssyndrome in non-obese diabetic mice.
   J Oral Pathol Med. 2012 May 29.
11.Gao D, Ning N, Hao G, Niu X.
   Pioglitazone attenuates vascular fibrosis in spontaneously hypertensive rats.
   PPAR Res. 2012;2012:856426.
12.Yuan Y, Guo-Qing P, Yan T, Hong-Lin Y, Gong-Hua H, Cai-Gao Z.
   A study of PKM2, PFK-1, and ANT1 expressions in cervical biopsy tissues in China.
   Med Oncol. 2012 Jan 8.
13.Ren J, Li W, Liu H, Yan L, Jiao W, Li D, Tang Y, Gu G, Xu Z.
   Overexpression of Reptin in renal cell carcinoma contributes to tumor malignancies and
   itsinhibition triggers senescence of cancer cells.
   Urol Oncol. 2012 Feb 16.
14.Chen X, Xu Z, Li L, Chen H, Zhou R.
   Identification of conserved surface proteins as novel antigenic vaccine candidates of
   Actinobacillus pleuropneumoniae.
   J Microbiol. 2012 Dec;50(6):978-86. doi: 10.1007/s12275-012-2214-2. Epub 2012 Dec 30.
15.Liu C, Chen Z, Tan C, Liu W, Xu Z, Zhou R, Chen H.
   Immunogenic characterization of outer membrane porins OmpC and OmpF of porcine
   extraintestinal pathogenicEscherichia coli.
   FEMS Microbiol Lett. 2012 Dec;337(2):104-11. doi: 10.1111/1574-6968.12013.
   Epub 2012 Oct 24.
16.Cai C, Wang C, Ji W, Liu B, Kang Y, Hu Z, Jiang Z.
   Knockdown of hepatic aquaglyceroporin-9 alleviates high fat diet-induced non-alcoholic
   fatty liver disease in rats.
   Int Immunopharmacol. 2013 Mar;15(3):550-6. doi: 10.1016/j.intimp.2013.01.020.
   Epub 2013 Feb 14.
17.Su L, Woodard RW, Chen J, Wu J.
   Extracellular location of Thermobifida fusca cutinase expressed in Escherichia coli
   BL21(DE3) without mediationof a signal peptide.
   Appl Environ Microbiol. 2013 Apr 19.
18.Ruan Q, Liu F, Gao Z, Kong D, Hu X, Shi D, Bao Z, Yu Z.
   The anti-inflamm-aging and hepatoprotective effects of huperzine A in
   D-galactose-treated rats.
   Mech Ageing Dev. 2013 Mar;134(3-4):89-97. doi: 10.1016/j.mad.2012.12.005.
   Epub 2013 Jan 8.
19.Chen XM, Xia J, Zhou T, Yuan Q, Zhang WF, Hu CP, Li YJ, Jiang JL.
   Involvement of DDAH/ADMA pathway in the pathogenesis of rheumatoid arthritis in rats.
   Int Immunopharmacol. 2013 Jun;16(2):322-31. doi: 10.1016/j.intimp.2013.04.009.
   Epub 2013 Apr 22.
20.Zhao F, Wu Y, Guo L, Li X, Han J, Chen Y, Ge Y.
   Using proteomics platform to develop a potential immunoassay method of royal jelly
   freshness.
   European Food Research and Technology. May 2013, Volume 236, Issue 5, pp 799-815.
21.Chen L, Zhou JP, Kuang DB, Tang J, Li YJ, Chen XP.
   4-HNE Increases Intracellular ADMA Levels in Cultured HUVECs: Evidence for
   miR-21-Dependent Mechanisms.
   PLoS One. 2013 May 22;8(5):e64148. doi: 10.1371/journal.pone.0064148. Print 2013.
22.Tang H, Ge L, Shao W, Qiu Y, Cui D.
   Effect of targeted silencing of hTERT mRNA by lentivirus-mediated siRNA on A549 lung
   cancer cells in vitro.
   Mol Biol Rep. 2013 Jan;40(1):605-16. doi: 10.1007/s11033-012-2099-5. Epub 2012 Oct 9.
23.Pan X, Wei Z, Wang H, Yu L, Liang X.
   Effects of dietary tryptophan on protein metabolism and related gene expression in
   Yangzhou goslings under different feeding regimens.
   Poult Sci. 2013 Dec;92(12):3196-204. doi: 10.3382/ps.2012-02953.
24.Mou XZ, Lin J, Chen JY, Li YF, Wu XX, Xiang BY, Li CY, Ma JM, Xiang C.
   Menstrual blood-derived mesenchymal stem cells differentiate into functional hepatocyte-
   like cells.
   J Zhejiang Univ Sci B. 2013 Nov;14(11):961-72. doi: 10.1631/jzus.B1300081.
25.Zhou H, Shi R, Wei M, Zheng WL, Zhou JY1, Ma WL.
   The expression and clinical significance of HERC4 in breast cancer.
   Cancer Cell Int. 2013 Nov 14;13(1):113. doi: 10.1186/1475-2867-13-113.
26.Xing X, Yang J, Yang X, Wei Y, Zhu L, Gao D, Li M.
   IL-17A induces endothelial inflammation in systemic sclerosis via the ERK signaling
   pathway.
   PLoS One. 2013 Dec 23;8(12):e85032. doi: 10.1371/journal.pone.0085032. eCollection
   2013.
27.Han Y, Wang S, Zhang Z, Ma X, Li W, Zhang X, Deng J, Wei H, Li Z, Zhang XE, Cui Z.
   In vivo imaging of protein-protein and RNA-protein interactions using novel far-red
   fluorescence complementation systems.
   Nucleic Acids Res. 2014;42(13):e103. doi: 10.1093/nar/gku408. Epub 2014 May 9.
28.Yin Y, Qi F, Song Z, Zhang B, Teng J.
   Ferulic acid combined with astragaloside IV protects against vascular endothelial
   dysfunction in diabetic rats.
   Biosci Trends. 2014 Aug;8(4):217-26.
29.Zhang XW, Zhang BY, Wang SW, Gong DJ, Han L, Xu ZY, Liu XH.
   Twist-related protein 1 negatively regulated osteoblastic transdifferentiation of human
   aortic valve interstitial cells by directly inhibiting runt-related transcription factor
   2.
   J Thorac Cardiovasc Surg. 2014 Oct;148(4):1700-1708.e1. doi: 10.1016/j.jtcvs.
   2014.02.084. Epub 2014 Mar 4.
30.Shi F, Xu Z, Chen H, Wang X, Cui J, Zhang P, Zhang P, Xie X.
   A Monoclonal Antibody Against PMEL.
   Monoclon Antib Immunodiagn Immunother. 2014 Oct;33(5):354-60. doi: 10.1089/mab.
   2013.0094. Epub 2014 Aug 13.
31.Lin X, Wu L, Zhang Z, Yang R, Guan Q, Hou X, Wu Q.
   MiR-335-5p promotes chondrogenesis in mouse mesenchymal stem cells and is regulated
   through two positive feedback loops.
   J Bone Miner Res. 2014 Jul;29(7):1575-85. doi: 10.1002/jbmr.2163.
32.Wu CP, Zhou L, Gong HL, Du HD, Tian J, Sun S, Li JY.
   Establishment and characterization of a novel HPV-negative laryngeal squamous cell
   carcinoma cell line, FD-LSC-1, with missense and nonsense mutations of TP53 in the
   DNA-binding domain.
   Cancer Lett. 2014 Jan 1;342(1):92-103. doi: 10.1016/j.canlet.2013.08.041. Epub 2013
   Aug 31.
33.Lin X, Wang Y, Zhang S, Zhu Z, Zhou YJ, Yang F, Sun W, Wang X, Zhao ZK.
   Functional integration of multiple genes into the genome of the oleaginous yeast
   Rhodosporidium toruloides.
   FEMS Yeast Res. 2014 Jun;14(4):547-55. doi: 10.1111/1567-1364.12140. Epub 2014 Feb
   17.
34.Zhang RP, Liu HH, Li QQ, Wang Y, Liu JY, Hu JW, Yan XP, Gou H, Li L, Wang JW.
   Gene expression patterns, and protein metabolic and histological analyses for
   muscle development in Peking duck.
   Poult Sci. 2014 Oct 11. pii: PS4145.
35.Li H, Jin G, Zhu P, Zou L, Shi J, Yi X, Zhang X, Tian M, Qin J.
   Upregulation of Lhx8 increase VAChT expression and ACh release in neuronal cell line
   SHSY5Y.
   Neurosci Lett. 2014 Jan 24;559:184-8. doi: 10.1016/j.neulet.2013.11.047. Epub 2013 Dec
   4.
36.Zeng Y, Xiang J, Lu Y, Chen Y, Wang T, Gong G, Wang L, Li X, Chen S, Sha Z.
   sghC1q, A Novel C1q Family Member from Half-smooth Tongue Sole (Cynoglossus
   semilaevis): identification, expression and analysis of antibacterial and antiviral
   activities.
   Dev Comp Immunol. 2014 Oct 10. pii: S0145-305X(14)00242-0. doi: 10.1016/j.dci.
   2014.09.010.
37.Tian Z, Sutton BJ, Zhang X.
   Distribution of rat neonatal Fc receptor in the principal organs of neonatal and
   pubertal rats.
   J Recept Signal Transduct Res. 2014 Apr;34(2):137-42. doi: 10.3109/10799893.
   2013.865745. Epub 2013 Dec 4.