Immunohistochemical localization of epidermal growth factor and acidic and basic fibroblast growth factors in postnatal developing and adult rat lungs.
Immunohistochemical localization of epidermal growth factor and acidic and basic fibroblast growth factors in postnatal developing and adult rat lungs.
复制标题
出生后发育和成年大鼠肺中表皮生长因子以及酸性和碱性成纤维细胞生长因子的免疫组织化学定位。
DOI：
10.1165/ajrcmb/7.2.230
复制
发表时间：
1992
影响因子：
6.4
通讯作者：
Khosla,J
中科院分区：
文献类型：
--
作者：
Sannes,PL;Burch,KK;Khosla,J
材料与方法组织制备无病毒抗体的1、5、10和18日龄大鼠和成年大鼠（200+ g）（Charles River Breeding Laboratories, Wilmington， MA）通过腹腔注射戊巴比妥钠安乐死，然后用酸性酒精（70%乙醇，1%盐酸）、2%多聚甲醛（0.1 M磷酸盐缓冲）或Bouin固定液注入肺部固定。15分钟后，修整肺部，再固定15分钟，脱水，常规处理石蜡包埋。5 - 7微米的切片被切割并安装在镀铬铝玻片上，并在室温下保存。兔抗小鼠EGF（受体级）血清购自Collaborative Research （Bedford， MA）或Upstate Biotechnology （Lake Placid， NY）。抗原纯化采用DEAE离子交换层析法。兔抗牛aFGF血清购自Upstate Biotechnology。抗原采用DEAE-blue层析纯化。根据制造商的说法，抗血清与bFGF有10%的交叉反应性。兔抗牛aFGF leu 60-leu 98片段（aFGFfr）血清购自Upstate Biotechnology。抗原是一种合成肽，代表aFGF中与bFGF同源性最少的部分。制造商指定aFGFfr与bFGF无交叉反应性。牛bFGF单克隆抗体购自Upstate Biotechnology。硫酸铵沉淀抗原经Sephacryl S-300和DEAE-52层析纯化。该抗体由P3-X63-Ag8-U1 x BALB/c脾细胞产生。制造商报告与aFGF无交叉反应。
Materials and MethodsTissue Preparations Virus antibody-free rats of 1, 5, 10, and 18 days of postnatal age and adult rats (200+ g)(Charles River Breeding Laboratories, Wilmington, MA) were euthanized by intraperitoneal injection with sodium pentobarbital, and lungs fixed by insufflation with either acid alcohol (70% ethanol, 1% hydrochloric acid), 2% paraformaldehyde (buffered with 0.1 M phosphate), or Bouin's fixative. After 15 min, the lungs were trimmed and fixed an additional 15 min, dehydrated, and processed routinely for paraffin embedment. Five-to sevenmicron sections were cut and mounted on chrome alumcoated glass slides and stored at room temperature.Immunostaining Preparations Rabbit antiserum to mouse EGF (receptor grade) was purchased from Collaborative Research (Bedford, MA) or from Upstate Biotechnology (Lake Placid, NY). Antigen purification was by DEAE ion-exchange chromatography. Rabbit antiserum to bovine aFGF was purchased from Upstate Biotechnology. Antigen purification was by DEAE-blue chromatography. According to the manufacturer, the antiserum had a 10% cross-reactivity with bFGF. Rabbit antiserum to the leu 60-leu 98 fragment of bovine aFGF (aFGFfr) was purchased from Upstate Biotechnology. The antigen was a synthetic peptide representing that portion of aFGF that has the least homology with bFGF. The manufacturer specified that the aFGFfr had no cross-reactivity with bFGF. Monoclonal antibodies to bovine bFGF were purchased from Upstate Biotechnology. The ammonium sulfate-precipitated antigen was purified by Sephacryl S-300 and DEAE-52 chromatography. The antibody was produced by P3-X63-Ag8-U1 x BALB/c spleen cells. The manufacturer reported no cross-reactivity with aFGF.