Biotin Conjugated Human CD27/TNFRSF7 Recombinant Rabbit Monoclonal Antibody [PSH08-33]
cat.: HA722976B
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human
Applications: ELISA(Det), ELISA
Clonality: Monoclonal
Clone number: PSH08-33
Form: Liquid
Storage condition: Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term.
Storage buffer: PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% ProClin300.
Concentration: 1ug/ul
Purification: Protein A affinity purified.
Isotype: IgG
Immunogen: Recombinant protein within Human CD27 aa 20-191 (HA210880).
Positive control: Recombinant Human CD27 protein (HA210880).
Subcellular location: Cell membrane.
Recommended Dilutions:
  ELISA(Det)
  ELISA
Use at an assay dependent concentration. Can be paired for Sandwich ELISA with Rabbit monoclonal [PSH08-31] to Human CD27 antibody (Capture) (HA722973) or Rabbit monoclonal [PSH08-32] to Human CD27 antibody (Capture) (HA722974) and recombinant Human CD27 protein (HA210880) as the standard. The reference range value is 12.3-4,000 pg/ml.
Use at an assay dependent concentration.
Uniprot #: SwissProt: P26842 Human
Alternative names: CD 27 CD27 CD27 antigen CD27 molecule CD27_HUMAN CD27L receptor LPFS2 MGC20393 OTTHUMP00000238557 S152 T cell activation antigen CD27 T cell antivation antigen S152 T-cell activation antigen CD27 T14 TNFRSF 7 TNFRSF7 TNFSF7 Tp 55 Tp55 Tumor necrosis factor receptor superfamily member 7
Images
HA722976B_1.png Fig1: Sandwich ELISA analysis of Human CD27 matched pair antibodies

Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA722973) diluted in carbonate/bicarbonate buffer, at a concentration of 2 µg/ml overnight at 4℃. Wells of the plate were washed, blocked with 150 µl 0.05% tween-20 1% BSA blocking buffer, and incubated with serial diluted Recombinant Human CD27 protein (HA210880) starting from 4000 pg/ml to 0 pg/ml and detect antibody (HA722976B, 0.2 µg/ml) for 1 hour at 30℃ with shaking. Then the plate was washed and incubated with 100 µl per well of SA-HRP for 0.5 hour at 30℃ with shaking. Detection was performed using an Ultra TMB Substrate for 10 minutes at room temperature in the dark. The reaction was stopped with sulfuric acid and absorbances were read on a spectrophotometer at 450 nm.
HA722976B_2.png Fig2: Sandwich ELISA analysis of Human CD27 matched pair antibodies

Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA722974) diluted in carbonate/bicarbonate buffer, at a concentration of 2 µg/ml overnight at 4℃. Wells of the plate were washed, blocked with 150 µl 0.05% tween-20 1% BSA blocking buffer, and incubated with serial diluted Recombinant Human CD27 protein (HA210880) starting from 4000 pg/ml to 0 pg/ml and detect antibody (HA722976B, 0.2 µg/ml) for 1 hour at 30℃ with shaking. Then the plate was washed and incubated with 100 µl per well of SA-HRP for 0.5 hour at 30℃ with shaking. Detection was performed using an Ultra TMB Substrate for 10 minutes at room temperature in the dark. The reaction was stopped with sulfuric acid and absorbances were read on a spectrophotometer at 450 nm.
HA722976B_3.png Fig3: Interpolated concentrations of native CD27 in human cell culture supernatant samples.

Interpolated concentration of native CD27 was measured in duplicate at different sample concentrations and interpolated from the CD27 standard curves. Undiluted samples were 50% cell supernatant. The interpolated dilution factor corrected values were plotted (mean +/- SD, n=2). The mean CD27 concentration was determined to be 3084 pg/ml in Daudi and 3995 pg/ml in Raji in cell culture supernatant, undetectable in MCF7 cell culture supernatant.
HA722976B_4.png Fig4: Interpolated concentrations of native CD27 in human cell culture supernatant samples.

Interpolated concentration of native CD27 was measured in duplicate at different sample concentrations and interpolated from the CD27 standard curves. Undiluted samples were 100% cell supernatant. The interpolated dilution factor corrected values were plotted (mean +/- SD, n=2). The mean CD27 concentration was determined to be 3041 pg/mL in Daudi and 3777 pg/ml in Raji in cell culture supernatant, undetectable in MCF7 cell culture supernatant.
HA722976B_5.png Fig5: Interpolated concentrations of spiked CD27 in cell culture media samples.

The concentrations of CD27 were measured in duplicates, interpolated from the CD27 standard curves and corrected for sample dilution. Undiluted samples are as follows: cell culture media 50%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2).
HA722976B_6.png Fig6: Interpolated concentrations of spiked CD27 in cell culture media samples.

The concentrations of CD27 were measured in duplicates, interpolated from the CD27 standard curves and corrected for sample dilution. Undiluted samples are as follows: cell culture media 50%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2).
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.