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CD86 Polyclonal Antibody

Human U937 Cells were fixed with 2% PFA (10min at room temperature). The cells were then incubated in 1X PBS containing 0.5% BSA + 10% goat serum to block non-specific protein-protein interactions for 15 min at room temperature. Staining was performed with CD86 Polyclonal Antibody, Unconjugated (bs-1035R) at 1:100 for 30 min on ice. A Goat Anti-rabbit IgG-PE secondary antibody was used for a 30 min incubation on ice. Primary antibody staining (green) is compared to unstained cells (dark blue), secondary antibody only (light blue) and isotype control (bs-0295P; orange).
Human U937 Cells were fixed with 2% PFA (10min at room temperature). The cells were then incubated in 1X PBS containing 0.5% BSA + 10% goat serum to block non-specific protein-protein interactions for 15 min at room temperature. Staining was performed with CD86 Polyclonal Antibody, Unconjugated (bs-1035R) at 1:100 for 30 min on ice. A Goat Anti-rabbit IgG-PE secondary antibody was used for a 30 min incubation on ice. Primary antibody staining (green) is compared to unstained cells (dark blue), secondary antibody only (light blue) and isotype control (bs-0295P; orange).
Raji cell lysates probed with CD86 Polyclonal Antibody, Unconjugated (bs-1035R) at 1:300 overnight at 4\u02daC. Followed by conjugation to secondary antibody (bs-0295G-HRP) at 1:500 for 90 min at 37\u02daC.
Raji cell lysates probed with CD86 Polyclonal Antibody, Unconjugated (bs-1035R) at 1:300 overnight at 4\u02daC. Followed by conjugation to secondary antibody (bs-0295G-HRP) at 1:500 for 90 min at 37\u02daC.
Mouse Brain cells probed with CD86 Polyclonal Antibody, PE Conjugated (bs-1035R-PE; green) at 1:50 compared to isotype control (bs-0295P-PE; orange).
Mouse Brain cells probed with CD86 Polyclonal Antibody, PE Conjugated (bs-1035R-PE; green) at 1:50 compared to isotype control (bs-0295P-PE; orange).
L1 rat brain lysates L2 rat heart lysates probed with Anti CD86\/B7-2 Polyclonal Antibody, Unconjugated (bs-1035R) at 1:200 overnight at 4\u02daC. Followed by conjugation to secondary antibody (bs-0295G-HRP) at 1:3000 for 90 min at RT. Predicted band 34kD. Observed band size:34kD.
L1 rat brain lysates L2 rat heart lysates probed with Anti CD86\/B7-2 Polyclonal Antibody, Unconjugated (bs-1035R) at 1:200 overnight at 4\u02daC. Followed by conjugation to secondary antibody (bs-0295G-HRP) at 1:3000 for 90 min at RT. Predicted band 34kD. Observed band size:34kD.
Mouse BV-2 Cells were fixed with 4% PFA (10min at room temperature) and then permeabilized with 0.1% PBS-Tween for 20 min at room temperature. The cells were then incubated in 1X PBS \/ 2% BSA \/ 10% goat serum to block non-specific protein-protein interactions for 20 min at 37\u00b0C. The cells were stained with CD86 Polyclonal Antibody Unconjugated (bs-1035R; green) at 1:100 for 90 min at 37\u00b0C. A FITC-conjugated Goat Anti-Rabbit IgG antibody (bs-0295G-FITC) was incubated at 37\u00b0C for 90 minutes. DAPI (5ug\/ml; blue) was used to stain the cell nuclei.
Mouse BV-2 Cells were fixed with 4% PFA (10min at room temperature) and then permeabilized with 0.1% PBS-Tween for 20 min at room temperature. The cells were then incubated in 1X PBS \/ 2% BSA \/ 10% goat serum to block non-specific protein-protein interactions for 20 min at 37\u00b0C. The cells were stained with CD86 Polyclonal Antibody Unconjugated (bs-1035R; green) at 1:100 for 90 min at 37\u00b0C. A FITC-conjugated Goat Anti-Rabbit IgG antibody (bs-0295G-FITC) was incubated at 37\u00b0C for 90 minutes. DAPI (5ug\/ml; blue) was used to stain the cell nuclei.
Lane 1: Human HepG2 cell lysates; Lane 2: Human U937 cell lysates; Lane 3: Rat Spleen lysates; Lane 4: Mouse Spleen lysates; Lane 5: Human HL-60 cell lysates probed with CD86 Polyclonal Antibody, Unconjugated (bs-1035R) at 1:1000 dilution and 4\u02daC overnight incubation. Followed by conjugated secondary antibody incubation at 1:20000 for 60 min at 37\u02daC.
Lane 1: Human HepG2 cell lysates; Lane 2: Human U937 cell lysates; Lane 3: Rat Spleen lysates; Lane 4: Mouse Spleen lysates; Lane 5: Human HL-60 cell lysates probed with CD86 Polyclonal Antibody, Unconjugated (bs-1035R) at 1:1000 dilution and 4\u02daC overnight incubation. Followed by conjugated secondary antibody incubation at 1:20000 for 60 min at 37\u02daC.
The Daudi(H) cells were incubated in 5%BSA to block non-specific protein-protein interactions (30 min at r.t.). Primary Antibody (green): Rabbit Anti-CD86 antibody (bs-1035R): 1 \u03bcg\/10^6 cells; Secondary Antibody (white blue): Goat anti-Rabbit IgG-FITC (bs-60295G-FITC): 1 \u03bcg\/test. Isotype Control (orange): Rabbit IgG (bs-0295P). Blank control (black): PBS. Acquisition of 20,000 events was performed.
The Daudi(H) cells were incubated in 5%BSA to block non-specific protein-protein interactions (30 min at r.t.). Primary Antibody (green): Rabbit Anti-CD86 antibody (bs-1035R): 1 \u03bcg\/10^6 cells; Secondary Antibody (white blue): Goat anti-Rabbit IgG-FITC (bs-60295G-FITC): 1 \u03bcg\/test. Isotype Control (orange): Rabbit IgG (bs-0295P). Blank control (black): PBS. Acquisition of 20,000 events was performed.
The MJ (H) cells were incubated in 5%BSA to block non-specific protein-protein interactions (30 min at r.t.) , followed by secondary antibody incubation for 40 min at room temperature. Primary Antibody (green): Rabbit Anti-CD86 antibody (bs-1035R): 1 \u03bcg\/10^6 cells; Isotype Control (orange): Rabbit IgG (bs-0295P). Blank control (black): PBS. Acquisition of 20,000 events was performed.
The MJ (H) cells were incubated in 5%BSA to block non-specific protein-protein interactions (30 min at r.t.) , followed by secondary antibody incubation for 40 min at room temperature. Primary Antibody (green): Rabbit Anti-CD86 antibody (bs-1035R): 1 \u03bcg\/10^6 cells; Isotype Control (orange): Rabbit IgG (bs-0295P). Blank control (black): PBS. Acquisition of 20,000 events was performed.
4% Paraformaldehyde-fixed Daudi (H) cell; Antibody incubation with (CD86) polyclonal Antibody, unconjugated (bs-1035R) 1:100, 90 min at 37\u00b0C; followed by conjugated Goat Anti-Rabbit IgG antibody (green, bs-40295G-FITC) at 37\u00b0C for 90 min, DAPI (blue, C02-04002) was used to stain the cell nuclei. PBS instead of the primary antibody was used as the blank control.
4% Paraformaldehyde-fixed Daudi (H) cell; Antibody incubation with (CD86) polyclonal Antibody, unconjugated (bs-1035R) 1:100, 90 min at 37\u00b0C; followed by conjugated Goat Anti-Rabbit IgG antibody (green, bs-40295G-FITC) at 37\u00b0C for 90 min, DAPI (blue, C02-04002) was used to stain the cell nuclei. PBS instead of the primary antibody was used as the blank control.
25 ug total protein per lane of various lysates (see on figure) probed with CD86 polyclonal antibody, unconjugated (bs-1035R) at 1:1000 dilution and 4\u00b0C overnight incubation. Followed by conjugated secondary antibody incubation at r.t. for 60 min.
25 ug total protein per lane of various lysates (see on figure) probed with CD86 polyclonal antibody, unconjugated (bs-1035R) at 1:1000 dilution and 4\u00b0C overnight incubation. Followed by conjugated secondary antibody incubation at r.t. for 60 min.
Bioss unconjugated antibody vial

Applications

  • WB
  • ELISA
  • FCM
  • IHC-P
  • IHC-F
  • IF(IHC-P)
  • IF(IHC-F)
  • IF(ICC)

Reactivity

  • Human
  • Mouse
  • Rat
Overview
Catalog # bs-1035R-100uL
Product Name CD86 Polyclonal Antibody
Applications
WB ELISA FCM IHC-P IHC-F IF(IHC-P) IF(IHC-F) IF(ICC)
Reactivity Human, Mouse, Rat
Specifications
Conjugation Unconjugated
Host Rabbit
Source KLH conjugated synthetic peptide derived from rat CD86
Immunogen Range 140-175/313
Clonality Polyclonal
Isotype IgG
Concentration 1ug/ul
Purification Purified by Protein A.
Storage Buffer 0.01M TBS(pH7.4) with 1% BSA, 0.02% Proclin300 and 50% Glycerol.
Storage Condition Shipped at 4C. Store at -20C for one year. Avoid repeated freeze/thaw cycles.
Target
Gene ID 56822
Swiss Prot O35531
Subcellular location Cell membrane
Synonyms B7-2, Cd86 antigen; Cd86; Membrane glycoprotein
Background This gene encodes a type I membrane protein that is a member of the immunoglobulin superfamily. This protein is expressed by antigen-presenting cells, and it is the ligand for two proteins at the cell surface of T cells, CD28 antigen and cytotoxic T-lymphocyte-associated protein 4. Binding of this protein with CD28 antigen is a costimulatory signal for activation of the T-cell. Binding of this protein with cytotoxic T-lymphocyte-associated protein 4 negatively regulates T-cell activation and diminishes the immune response. Alternative splicing results in several transcript variants encoding different isoforms.
Application Dilution
WB 1:300-5000
ELISA 1:500-1000
FCM 1:20-100
IHC-P 1:200-400
IHC-F 1:100-500
IF(IHC-P) 1:50-200
IF(IHC-F) 1:50-200
IF(ICC) 1:50-200

Frequently asked questions

Which applications is CD86 Polyclonal Antibody (bs-1035R) validated for, and at what dilution?

bs-1035R is validated for WB (1:300-5000); ELISA (1:500-1000); FCM (1:20-100); IHC-P (1:200-400); IHC-F (1:100-500); IF(IHC-P) (1:50-200); IF(IHC-F) (1:50-200); IF(ICC) (1:50-200). These are recommended starting dilutions; optimal dilutions should be determined by the end user.

Which species does CD86 Polyclonal Antibody (bs-1035R) react with?

Tested reactivity: Human, Mouse, Rat.

How should bs-1035R be stored?

Shipped at 4C. Store at -20C for one year. Avoid repeated freeze/thaw cycles. Storage buffer: 0.01M TBS(pH7.4) with 1% BSA, 0.02% Proclin300 and 50% Glycerol.

What immunogen was used to raise CD86 Polyclonal Antibody (bs-1035R)?

KLH conjugated synthetic peptide derived from rat CD86 Immunogen range: 140-175/313.

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