TNFR2 Recombinant Rabbit Monoclonal Antibody

• Catalog Number: orb612211
• Category: Antibodies
• Description: TNFR2 Recombinant Rabbit Monoclonal Antibody
• Species/Host: Rabbit
• Clonality: Recombinant
• Clone Number: 7D3
• Tested applications: FC, ICC, IF, IHC-Fr, IHC-P, WB
• Predicted Reactivity: Rat
• Reactivity: Human, Mouse
• Isotype: IgG
• Immunogen: KLH conjugated synthetic peptide derived from human TNFR2
• Concentration: 1mg/ml
• Dilution range: WB=1:500-2000, IHC-P=1:50-200, IHC-F=1:50-200, ICC/IF=1:50, IF=1:50-200, Flow-Cyt=1:50
• Form/Appearance: Liquid
• Conjugation: Unconjugated
• MW: 46 kDa
• Target: TNFRSF1B
• UniProt ID: P20333
• Storage: Maintain refrigerated at 2-8°C for up to 2 weeks. For long term storage store at -20°C in small aliquots to prevent freeze-thaw cycles.
• Buffer/Preservatives: 0.01M TBS (pH7.4) with 1% rAlbumin, 0.02% Proclin300 and 50% Glycerol.
• Alternative names: TNF Receptor II; CD120b; p75; p75TNFR; p80 TNF alp
• Note: For research use only
• Expiration Date: 12 months from date of receipt.

Blank control: HL-60. Primary Antibody (green line): Rabbit Anti-TNF Receptor II antibody (orb612211), dilution: 1:50, Isotype Control Antibody (orange line): Rabbit IgG. Secondary Antibody: Goat anti-rabbit IgG-AF488, dilution: 1:1000. Protocol, The cells were fixed with 4% PFA (10 min at room temperature) and then permeabilized with 0.1% PBST for 20 min at room temperature. The cells were then incubated in 5% BSA to block non-specific protein-protein interactions for 30 min at room temperature. Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20000 events was performed.

Hela cell, 4% Paraformaldehyde-fixed, Triton X-100 at room temperature for 20 min, Blocking buffer (normal goat serum) at 37°C for 20 min, Antibody incubation with (TNF Receptor II) monoclonal Antibody, Unconjugated (orb612211) 1:50, 90 minutes at 37°C, followed by a conjugated Goat Anti-Rabbit IgG antibody at 37°C for 90 minutes, DAPI (blue) was used to stain the cell nuclei.

MCF-7 cell, 4% Paraformaldehyde-fixed, Triton X-100 at room temperature for 20 min, Blocking buffer (normal goat serum) at 37°C for 20 min, Antibody incubation with (TNF Receptor II) monoclonal Antibody, Unconjugated (orb612211) 1:50, 90 minutes at 37°C, followed by a conjugated Goat Anti-Rabbit IgG antibody at 37°C for 90 minutes, DAPI (blue) was used to stain the cell nuclei.

Paraformaldehyde-fixed, paraffin embedded (human kidney tissue), Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min, Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes, Blocking buffer (normal goat serum) at 37°C for 30 min, Antibody incubation with (TNFR2) Monoclonal Antibody, Unconjugated (orb612211) at 1:50 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.

Paraformaldehyde-fixed, paraffin embedded (human uterus tissue), Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min, Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes, Blocking buffer (normal goat serum) at 37°C for 30 min, Antibody incubation with (TNFR2) Monoclonal Antibody, Unconjugated (orb612211) at 1:50 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.

Paraformaldehyde-fixed, paraffin embedded (mouse kidney tissue), Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min, Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes, Blocking buffer (normal goat serum) at 37°C for 30 min, Antibody incubation with (TNFR2) Monoclonal Antibody, Unconjugated (orb612211) at 1:50 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.

Sample: Lane 1: Human THP-1 cell Lysates, Lane 2: Human K562 cell Lysates, Lane 3: Human U937 cell Lysates, Lane 4: Human U87MG cell Lysates, Lane 5: Human HUVEC cell Lysates, Primary: Anti-TNFR2 (orb612211) at 1/1000 dilution, Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution, Predicted band size: 46kDa, Observed band size: 70 kDa.

Sample: Lane 1: MCF-7 cell lysate, Lane 2: Jurkat cell lysate, Lane 3: Hela cell lysate, Primary: Anti-TNFR2 (orb612211) at 1:500 dilution, Secondary: Goat Anti-Rabbit IgG - HRP at 1:5000 dilution, Predicted band size: 46 kD, Observed band size: 60 kD.

SW480 cell, 4% Paraformaldehyde-fixed, Triton X-100 at room temperature for 20 min, Blocking buffer (normal goat serum) at 37°C for 20 min, Antibody incubation with (TNF Receptor II) monoclonal Antibody, Unconjugated (orb612211) 1:50, 90 minutes at 37°C, followed by a conjugated Goat Anti-Rabbit IgG antibody at 37°C for 90 minutes, DAPI (blue) was used to stain the cell nuclei.