Rabbit polyclonal antibody to Cd86
WB, IHC-P, ICC/IF, FC
Human, Mouse, Rat, Dog, Pig, Cow, Sheep
This antigen of this antibody was klh conjugated synthetic peptide derived from the middle of rat cd86:140-175/313
Liquid, 0.01M TBS(pH7.4) with 1% BSA, 0.03% Proclin300 and 50% Glycerol.
Store at -20℃ for one year. Avoid repeated freeze/thaw cycles. The lyophilized antibody is stable at room temperature for at least one month and for greater than a year when kept at -20℃. When reconstituted in sterile pH 7.4 0.01M PBS or diluent of antibody the antibody is stable for at least two weeks at 2-4℃.
Fig1: Tissue/cell: BV-2 cell; 4% Paraformaldehyde-fixed; Triton X-100 at room temperature for 20 min; Blocking buffer (normal goat serum, C-0005) at 37℃ for 20 min; Antibody incubation with (CD86) Polyclonal Antibody, Unconjugated 1:200, 90 minutes at 37℃; followed by a conjugated Goat Anti-Rabbit IgG antibody (bs-0295G-FITC) at 37℃ for 90 minutes, DAPI (5ug/ml, blue, C-0033) was used to stain the cell nuclei.
Fig2: Sample:; Lymph node(Mouse)Cell Lysate at 40 ug; Primary: Anti-CD86 at 1/300 dilution; Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution; Predicted band size: 31 kD; Observed band size: 31 kD
Fig3: Tissue/cell: rat lung tissue; 4% Paraformaldehyde-fixed and paraffin-embedded;; Antigen retrieval: citrate buffer ( 0.01M, pH 6.0 ), Boiling bathing for 15min; Block endogenous peroxidase by 3% Hydrogen peroxide for 30min; Blocking buffer (normal goat serum,C-0005) at 37℃ for 20 min;; Incubation: Anti-CD86/B7-2 Polyclonal Antibody, Unconjugated 1:200, overnight at 4℃, followed by conjugation to the secondary antibody(SP-0023) and DAB(C-0010) staining
Fig4: Sample:; Brain(Rat) lysates at 30ug;; Heart(Rat) lysates, 30ug;; Primary: Anti-CD86/B7-2 at 1:200;; Secondary: HRP conjugated Goat Anti-Rabbit IgG(bs-0295G-HRP) at 1: 3000;; ECL excitated the fluorescence;; Predicted band size : 34kD; Observed band size : 34kD
Fig5: Blank control: U937(blue).; Primary Antibody: Rabbit Anti-CD86 antibody , Dilution: 1μg in 100 μL 1X PBS containing 0.5% BSA;; Isotype Control Antibody: Rabbit IgG (orange) ,used under the same conditions.; Secondary Antibody: Goat anti-rabbit IgG-PE(white blue), Dilution: 1:200 in 1 X PBS containing 0.5% BSA.; Protocol; The cells were fixed with 2% paraformaldehyde (10 min).Primary antibody ( 1μg /1x10^6 cells) were incubated for 30 min on the ice, followed by 1 X PBS containing 0.5% BSA + 10% goat serum (15 min) to block non-specific protein-protein interactions. Then the Goat Anti-rabbit IgG/PE antibody was added into the blocking buffer mentioned above to react with the primary antibody at 1/200 dilution for 30 min on ice. Acquisition of 20,000 events was performed.
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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"