Product Specifications:
Item# 1041: Recombinant HIV-1 IIIB Glycoprotein gp120 (CHO)
Concentration: See vial
Diluent: PBS
Purity: >95%
Stabilizer: None
Preservative: None
Storage: -75°C
Physical State: Frozen
Liquid Stability: At least 6 months at -75°C.
Applications: In-Vitro Diagnostics, CD4 Binding, T-Cell Activation.
Description: Full length Recombinant HIV-1 IIIB gp120 glycoprotein produced in the CHO Expression System.
Purification: This protein is purified by immunoaffinity chromatography to >95% purity as determined by SDS-PAGE, reduced.
Specificity: This protein binds to murine monoclonal antibodies of defined epitope specificity and human serum polyclonal antibodies in ELISA and Western ELISA.
Biological Activity: This protein binds to human T-cell receptor CD4 in ELISA and Western ELISA as determined by CD4/gp120/Anti-gp120 mAb-peroxidase capture ELISA. This protein activates human T-Lymphocytes (CD4+, CD4-), in vitro, as measured by RNA synthesis during G0 to G1 transition phase of antigen-binding competent cells.
Application and Instructions for use
Recommended concentrations for use are approximate values. A dose dependent response assay should be performed to determine the optimal concentration for use in specific applications. ELISA and Western ELISA require 10-100ng protein depending on the nature and affinity of the detection reagent. Human serum polyclonal antibodies yield titers of 1:1000 or greater at 100ng of solid phase protein under standard ELISA conditions.
Item#: 1041-P Peroxidase-Conjugated rgp120 HIV-1 IIIB
Application: Human diagnostics, CD4 binding, Drug screening, Human T-cell CD4 studies.
Safety Study of rgp120/HIV-1IIIB Vaccine
Recent evidence suggests that gp120 is the HIV-1 protein with the greatest potential as a vaccine against HIV-1 infection. The gp120 envelope protein may be produced by recombinant DNA technology, and studies have shown that the vaccine is capable of eliciting neutralizing antibody activity in both rodents and nonhuman primate species.
Official Study Title: A Phase I Study of the Safety and Immunogenicity of rgp120/HIV-1IIIB Vaccine in Healthy Adult Subjects (NOTE: Study Extended ONLY for Subjects Who Have Previously Received rgp120/HIV-1IIIB or rgp120/HIV-1MN on VEU 006 or VEU 006 Rollover Study)
Disease: HIV Infections
Treatment: Biological: rgp120/HIV-1IIIB and Biological: rgp120/HIV-1MN
ORIGINAL DESIGN: Twenty-eight subjects will be randomized to receive 100 or 300 mcg rgp120/HIV-1IIIB vaccine (gp120 vaccine) or matching placebo. For each dose level, 10 subjects will receive vaccine and four subjects will receive matching placebo. Injections are given intramuscularly at 0, 4, and 32 weeks. Each subject receiving treatment at the lower dose level must be monitored for unacceptable toxicity for at least 2 weeks following the initial immunization before his or her second dose is administered and before treatment at the higher dose level begins. Subjects are followed for at least 12 months.
AMENDED 11/17/93: Selected subjects from VEU 006 or VEU 006 Rollover study will receive two injections of MN rgp120 vaccine, administered 28 days apart beginning 10-16 months after their last injection. Eight additional clinic visits will be required. Subjects are followed for at least 6 months.
CHO- expressed HIV-1IIIB rgp120 (2.5 μg) and biotinylated cyclic V2-TH023 peptide (1 μg and 5 μg) were incubated with RPMI8866 cells
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多谢！
不知道能不能直接买到大鼠抗豚鼠血管IgG的ELISA试剂盒。
如果买不到这种试剂盒，用空白板的话（就是反应孔中没有抗原或者抗体包被），把自制的豚鼠血管抗原加到反应孔中进行孵育，能不能使抗原附着在孔壁上？这样的话就可以按照elisa的操作步骤进行大鼠血清检测了。
目前的设想是自制抗原，粉碎豚鼠的血管，制成悬液，通过反复多次的冻融，离心后收集上清液作为抗原（即豚鼠血管抗原）。
然后将抗原加入到elisa的反应孔中（这种是特制的空白板，就是反应孔中没有抗原或抗体包被）进行孵育，使豚鼠血管抗原附着在孔壁上，就是让反应孔充当固相载体，形成固相抗原。倒掉多余的抗原。
再加入待检测的大鼠血清，这样血清中的特异性抗豚鼠血管IgG就可以跟固相的抗原结合，形成固相抗原抗体复合物。
加入酶标的兔抗鼠或者羊抗鼠IgG,形成酶标的抗原抗体复合物。
然后就是一些显色步骤。
现在关键的问题就是抗原加入空白板，能不能形成固相抗原，如果不能跟孔壁附着的话，在后面洗涤的过程中就被洗掉了，那就没办法完成检测了。
请各位大侠给指条明路啊！！！小弟在此多谢了！！！
