CD80 Recombinant Rabbit Monoclonal Antibody [PSH18-13]
cat.: HA723976
| Product Type: | Recombinant Rabbit monoclonal IgG, primary antibodies |
|---|---|
| Species reactivity: | Mouse |
| Applications: | WB, IF-Cell, FC, IP |
| Clonality: | Monoclonal |
| Clone number: | PSH18-13 |
| Form: | Liquid |
| Storage condition: | Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term. |
| Storage buffer: | 1*PBS (pH7.4), 0.1% BSA, 40% Glycerol, 0.2% Proclean 950. |
| Concentration: | 1ug/ul |
| Purification: | Protein A affinity purified. |
| Molecular weight: | Predicted band size: 35 kDa |
| Isotype: | IgG |
| Immunogen: | Recombinant protein within mouse CD80 aa 1-246. |
| Positive control: | C2C12 cell lysate, RAW264.7 cell lysate, RAW264.7 treated with 1μg/mL LPS for 24 hours cell lysate, C2C12. |
| Subcellular location: | Cell membrane. |
| Recommended Dilutions:
WB IF-Cell FC IP |
1:5,000 1:100 1:1,000 1-2μg/sample |
| Uniprot #: | SwissProt: Q00609 Mouse |
| Alternative names: | Activation B7-1 antigen B lymphocyte activation antigen B7 B7 B7-1 B7-1 antigen B7.1 BB1 CD28 antigen ligand 1 CD28LG CD28LG1 CD80 CD80 antigen (CD28 antigen ligand 1, B7-1 antigen) CD80 antigen CD80 molecule Costimulatory factor CD80 costimulatory molecule variant IgV-CD80 CTLA-4 counter-receptor B7.1 LAB7 T-lymphocyte activation antigen CD80 |
Images
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Fig1:
Western blot analysis of CD80 on different lysates with Rabbit anti-CD80 antibody (HA723976) at 1/5,000 dilution. Lane 1: C2C12 cell lysate Lane 2: NIH/3T3 cell lysate (negative) Lysates/proteins at 20 µg/Lane. Predicted band size: 35 kDa Observed band size: 60 kDa Exposure time: 59 seconds; ECL: K1801; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA723976) at 1/5,000 dilution was used in primary antibody dilution (K1803) at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. |
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Fig2:
Western blot analysis of CD80 on different lysates with Rabbit anti-CD80 antibody (HA723976) at 1/5,000 dilution. Lane 1: RAW264.7 cell lysate Lane 2: RAW264.7 treated with 1μg/mL LPS for 24 hours cell lysate Lysates/proteins at 20 µg/Lane. Predicted band size: 35 kDa Observed band size: 60 kDa Exposure time: 3 minutes; ECL: K1801; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA723976) at 1/5,000 dilution was used in primary antibody dilution (K1803) at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. |
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Fig3:
Immunocytochemistry analysis of C2C12 (positive) and NIH/3T3 (negative) labeling CD80 with Rabbit anti-CD80 antibody (HA723976) at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-CD80 antibody (HA723976) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution. |
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Fig4:
Flow cytometric analysis of NIH/3T3 (left, negative) and C2C12 (right, positive) cells labeling CD80. Cells were fixed and permeabilized. Then stained with the primary antibody (HA723976, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black). |
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Fig5:
CD80 was immunoprecipitated from 0.2 mg C2C12 cell lysate with HA723976 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA723976 at 1/5,000 dilution. HRP Conjugated Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature. Lane 1: C2C12 cell lysate (input) Lane 2: HA723976 IP in C2C12 cell lysate Lane 3: Rabbit IgG instead of HA723976 in C2C12 cell lysate Blocking/Dilution buffer: primary antibody dilution (K1803) Exposure time: 20 seconds; ECL: K1801 |
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.