CD80 Recombinant Rabbit Monoclonal Antibody [PSH18-13] - BSA and Azide free
Catalog# HA751659
CD80 Recombinant Rabbit Monoclonal Antibody [PSH18-13] - BSA and Azide free
-
WB
-
IF-Cell
-
FC
-
IP
-
Mouse
-
HA723976
含抗保成分
-
unconjugated
Overview
Product Name
CD80 Recombinant Rabbit Monoclonal Antibody [PSH18-13] - BSA and Azide free
Antibody Type
Recombinant Rabbit monoclonal Antibody
Immunogen
Recombinant protein within mouse CD80 aa 1-246.
Species Reactivity
Mouse
Validated Applications
WB, IF-Cell, FC, IP
Molecular Weight
Predicted band size: 35 kDa
Positive Control
C2C12 cell lysate, RAW264.7 cell lysate, RAW264.7 treated with 1μg/mL LPS for 24 hours cell lysate, C2C12.
Conjugation
unconjugated
Clone Number
PSH18-13
Product Features
Form
Liquid
Concentration
1mg/ml
Storage Instructions
Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
Storage Buffer
1*PBS (pH7.4).
Isotype
IgG
Purification Method
Protein A affinity purified.
Application Dilution
-
WB
-
1:5,000
-
IF-Cell
-
1:100
-
FC
-
1:1,000
-
IP
-
1-2μg/sample
Target
Function
The Cluster of differentiation 80 (also CD80 and B7-1) is a B7, type I membrane protein in the immunoglobulin superfamily, with an extracellular immunoglobulin constant-like domain and a variable-like domain required for receptor binding. It is closely related to CD86, another B7 protein (B7-2), and often works in tandem. Both CD80 and CD86 interact with costimulatory receptors CD28, CTLA-4 (CD152) and the p75 neurotrophin receptor. CD80 can be found on the surface of various immune cells, including B-cells, monocytes, or T-cells, but most typically at antigen-presenting cells (APCs) such as dendritic cells. CD80 has a crucial role in modulating T-cell immune function as a checkpoint protein at the immunological synapse.
Background References
1. Sugiura D et al. PD-1 agonism by anti-CD80 inhibits T cell activation and alleviates autoimmunity. Nat Immunol. 2022 Mar
2. Tekguc M et al. Treg-expressed CTLA-4 depletes CD80/CD86 by trogocytosis, releasing free PD-L1 on antigen-presenting cells. Proc Natl Acad Sci U S A. 2021 Jul
Subcellular Location
Cell membrane.
UNIPROT
Synonyms
Activation B7-1 antigen antibody
B lymphocyte activation antigen B7 antibody
B7 antibody
B7-1 antibody
B7-1 antigen antibody
B7.1 antibody
BB1 antibody
CD28 antigen ligand 1 antibody
CD28LG antibody
CD28LG1 antibody
ExpandImages
-
☑ Relative expression (RE)
Western blot analysis of CD80 on different lysates with Rabbit anti-CD80 antibody (HA751659) 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 (HA751659) 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. -
☑ Cell treatment (CT)
Western blot analysis of CD80 on different lysates with Rabbit anti-CD80 antibody (HA751659) 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 (HA751659) 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. -
☑ Relative expression (RE)
Immunocytochemistry analysis of C2C12 (positive) and NIH/3T3 (negative) labeling CD80 with Rabbit anti-CD80 antibody (HA751659) 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 (HA751659) 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. -
☑ Relative expression (RE)
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 (HA751659, 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). -
CD80 was immunoprecipitated from 0.2 mg C2C12 cell lysate with HA751659 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA751659 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: HA751659 IP in C2C12 cell lysate
Lane 3: Rabbit IgG instead of HA751659 in C2C12 cell lysate
Blocking/Dilution buffer: primary antibody dilution (K1803)
Exposure time: 20 seconds; ECL: K1801
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"