Application
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 (ER1906-01) 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.Application
Fig2: Sample: Lymph node(Mouse)Cell Lysate at 40 ug Primary: Anti-CD86 (ER1906-01) at 1/300 dilution Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution Predicted band size: 31 kD Observed band size: 31 kDApplication
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(ER1906-01) 1:200, overnight at 4℃, followed by conjugation to the secondary antibody(SP-0023) and DAB(C-0010) stainingApplication
Fig4: Sample: Brain(Rat) lysates at 30ug; Heart(Rat) lysates, 30ug; Primary: Anti-CD86/B7-2 (ER1906-01) 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 : 34kDApplication
Fig5: Blank control: U937(blue). Primary Antibody: Rabbit Anti-CD86 antibody(ER1906-01), 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 (ER1906-01, 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.WB
(a–c) Representative western blot bands and analysis of CD86 and CD206 around the lesion area (n = 6).WB
(a–c) Representative western blot bands and analysis of CD86 and CD206 around the lesion area (n = 6).WB
(a–c) Representative western blot bands and analysis of CD86 and CD206 around the lesion area (n = 6).IF
(d and f) Immunofluorescence of CD86+/CD11b+ and CD206+/CD11b+ cells in the lesion area (n = 3, each sample was calculated from two separate fields of view).IF
(d and f) Immunofluorescence of CD86+/CD11b+ and CD206+/CD11b+ cells in the lesion area (n = 3, each sample was calculated from two separate fields of view).| Product Name | CD86 |
|---|---|
| Antibody Type | Primary Antibodies |
| Product description | Receptor involved in the costimulatory signal essential for T-lymphocyte proliferation and interleukin-2 production, by binding CD28 or CTLA-4. May play a critical role in the early events of T-cell activation and costimulation of naive T-cells, such as deciding between immunity and anergy that is made by T-cells within 24 hours after activation. Isoform 2 interferes with the formation of CD86 clusters, and thus acts as a negative regulator of T-cell activation. |
| Immunogen | KLH conjugated synthetic peptide derived from the middle of human CD86 140-175/313 |
| Clonality | Polyclonal |
|---|---|
| Isotype | IgG |
| Host Species | Rabbit |
| Tested Applications | FCICC/IFIHC-PWB |
| WB:1:500-2000: IHC-P:1:400-800: FC:1μg/Test: IF:1:100-500: ICC:1:100-500: |
|
| Species Reactivity | CowDogHumanMousePigRatSheep |
| Concentration | 1mg/ml |
| Alternative Names | CD28LG2
FUN1
LAB72
Activation B7 2 antigen
Activation B7-2 antigen 3
CD86_HUMAN
Activation B7-2 antigen
Activation B72 antigen
B lymphocyte activation antigen B7 2
B lymphocyte activation antigen B72
B-lymphocyte activation antigen B7-2 2
B-lymphocyte activation antigen B7-2
B7 2
B7 2 antigen
B7
B7-2
B7.2
B70
B72 antigen
BU63
CD 86
CD28 antigen ligand 2 2
CD28 antigen ligand 2
Cd28l2
CD28LG2
CD86 antigen (CD28 antigen ligand 2 B7-2 antigen) 1 2 CD86 antigen CD86 molecule CLS1 CTLA 4 counter receptor B7 2 CTLA 4 counter receptor B7.2 CTLA-4 counter-receptor B7.2 2 3 CTLA4 counter receptor B72 Early T-cell costimulatory molecule 1 ETC-1 FUN 1 LAB72 Ly-58 Ly58 MB7 MB7-2 Membrane glycoprotein MGC34413 T lymphocyte activation antigen CD86 T-lymphocyte activation antigen CD86 TS/A-2. |
|---|---|
| Molecular Weight(MW) | 31 KDa |
| Cellular Localization | Cell membrane; Single-pass type I membrane protein. |
| SwissProt ID | O35531 |
|---|
Application
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 (ER1906-01) 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.
Application
Fig2: Sample: Lymph node(Mouse)Cell Lysate at 40 ug Primary: Anti-CD86 (ER1906-01) at 1/300 dilution Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution Predicted band size: 31 kD Observed band size: 31 kD
Application
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(ER1906-01) 1:200, overnight at 4℃, followed by conjugation to the secondary antibody(SP-0023) and DAB(C-0010) staining
Application
Fig4: Sample: Brain(Rat) lysates at 30ug; Heart(Rat) lysates, 30ug; Primary: Anti-CD86/B7-2 (ER1906-01) 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
Application
Fig5: Blank control: U937(blue). Primary Antibody: Rabbit Anti-CD86 antibody(ER1906-01), 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 (ER1906-01, 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.
WB
(a–c) Representative western blot bands and analysis of CD86 and CD206 around the lesion area (n = 6).
WB
(a–c) Representative western blot bands and analysis of CD86 and CD206 around the lesion area (n = 6).
WB
(a–c) Representative western blot bands and analysis of CD86 and CD206 around the lesion area (n = 6).
IF
(d and f) Immunofluorescence of CD86+/CD11b+ and CD206+/CD11b+ cells in the lesion area (n = 3, each sample was calculated from two separate fields of view).
IF
(d and f) Immunofluorescence of CD86+/CD11b+ and CD206+/CD11b+ cells in the lesion area (n = 3, each sample was calculated from two separate fields of view).| Positive Control | Expressed by activated B-lymphocytes and monocytes. |
|---|---|
| Application Notes | WB:1:500-2000: IHC-P:1:400-800: FC:1μg/Test: IF:1:100-500: ICC:1:100-500: |
| Form | Liquid |
|---|---|
| Storage Instructions | 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℃. |
| Storage Buffer | 0.01M TBS(pH7.4) with 1% BSA, 0.03% Proclin300 and 50% Glycerol. |
