Human CD28 Recombinant Rabbit Monoclonal Antibody [PSH12-51] - BSA and Azide free (Detector)
Overview
Product Name
Human CD28 Recombinant Rabbit Monoclonal Antibody [PSH12-51] - BSA and Azide free (Detector)
Antibody Type
Recombinant Rabbit monoclonal Antibody
Immunogen
Recombinant protein within Human CD28 aa 19-152 (HA210920).
Species Reactivity
Human
Validated Applications
ELISA(Det)
Positive Control
Recombinant Human CD28 protein (HA210920).
Conjugation
unconjugated
Clone Number
PSH12-51
Product Features
Form
Liquid
Concentration
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
-
ELISA(Det)
-
Use at an assay dependent concentration. Can be paired for Sandwich ELISA with Rabbit monoclonal [PSH12-50] to Human CD28 antibody (Capture) (HA723457) and Recombinant Human CD28 protein (HA210920) as the standard. The reference range value is 39-5,000 pg/mL.
Target
Function
CD28 (Cluster of Differentiation 28) is a protein expressed on T cells that provides essential co-stimulatory signals required for T cell activation and survival. When T cells are stimulated through CD28 in conjunction with the T-cell receptor (TCR), it enhances the production of various interleukins, particularly IL-6. CD28 serves as a receptor for CD80 (B7.1) and CD86 (B7.2), proteins found on antigen-presenting cells (APCs). CD28 is the only B7 receptor consistently expressed on naive T cells. In the absence of CD28:B7 interaction, a naive T cell's TCR engagement with an MHC:antigen complex leads to anergy. CD28 is also expressed on bone marrow stromal cells, plasma cells, neutrophils, and eosinophils, although its function in these cells is not fully understood. Typically, CD28 is expressed on about 50% of CD8+ T cells and more than 80% of CD4+ T cells in humans. However, some T cells lose CD28 expression during activation, particularly antigen-experienced T cells, which can be re-activated independently of CD28. These CD28− T cells are often antigen-specific, terminally differentiated, and categorized as memory T cells (TMs). The proportion of CD28− T cells increases with age.
Background References
1. Cappell KM et al. A comparison of chimeric antigen receptors containing CD28 versus 4-1BB costimulatory domains. Nat Rev Clin Oncol. 2021 Nov
2. Lotze MT et al. CD28 co-stimulation: novel insights and applications in cancer immunotherapy. Nat Rev Immunol. 2024 Dec
Subcellular Location
Cell membrane; Cell surface.
UNIPROT
Synonyms
CD 28 antibody
CD28 antibody
CD28 antigen antibody
CD28 molecule antibody
CD28_HUMAN antibody
MGC138290 antibody
T cell antigen CD28 antibody
T cell specific surface glycoprotein antibody
T cell specific surface glycoprotein CD28 antibody
T-cell-specific surface glycoprotein CD28 antibody
ExpandImages
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![Sandwich ELISA analysis of Human CD28 matched pair antibodies<br /><br />Capture: <a href="/products/HA723457" style="font-weight: bold;text-decoration: underline;">HA723457</a>, Human CD28 Rabbit mAb [PSH12-50]<br />Detector: <a href="/products/HA723458" style="font-weight: bold;text-decoration: underline;">HA723458</a>, Human CD28 Rabbit mAb [PSH12-51]<br /><br />Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (<a href="/products/HA723457" style="font-weight: bold;text-decoration: underline;">HA723457</a>) diluted in carbonate/bicarbonate buffer, at a concentration of 2ug/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 CD28 protein (<a href="/products/HA210920" style="font-weight: bold;text-decoration: underline;">HA210920</a>) starting from 10,000 pg/ml to 0 pg/ml and detect antibody (<a href="/products/HA723458" style="font-weight: bold;text-decoration: underline;">HA723458</a>, Biotin, 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.](https://storage.huabio.cn/huabio/productImg/HA723458_1.png?v=20260401002256)
Sandwich ELISA analysis of Human CD28 matched pair antibodies
Capture: HA723457, Human CD28 Rabbit mAb [PSH12-50]
Detector: HA723458, Human CD28 Rabbit mAb [PSH12-51]
Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA723457) diluted in carbonate/bicarbonate buffer, at a concentration of 2ug/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 CD28 protein (HA210920) starting from 10,000 pg/ml to 0 pg/ml and detect antibody (HA723458, Biotin, 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. -
![Interpolated concentrations of native CD28 in human samples.<br /><br />Capture: <a href="/products/HA723457" style="font-weight: bold;text-decoration: underline;">HA723457</a>, Human CD28 Rabbit mAb [PSH12-50]<br />Detector: <a href="/products/HA723458" style="font-weight: bold;text-decoration: underline;">HA723458</a>, Human CD28 Rabbit mAb [PSH12-51]<br /><br />Interpolated concentration of native CD28 was measured in duplicate at different sample concentrations. Undiluted samples were 100% cell supernatant. The mean CD28 concentration was determined to be 275 pg/mL in Jurkat cell culture supernatant. There was no detectable signal in THP-1 cell culture supernatant.](https://storage.huabio.cn/huabio/productImg/HA723458_2.png?v=20260401002256)
Interpolated concentrations of native CD28 in human samples.
Capture: HA723457, Human CD28 Rabbit mAb [PSH12-50]
Detector: HA723458, Human CD28 Rabbit mAb [PSH12-51]
Interpolated concentration of native CD28 was measured in duplicate at different sample concentrations. Undiluted samples were 100% cell supernatant. The mean CD28 concentration was determined to be 275 pg/mL in Jurkat cell culture supernatant. There was no detectable signal in THP-1 cell culture supernatant. -
![Interpolated concentrations of native CD28 in Jurkat and THP-1 extract samples based on a 1000 µg/ml extract load.<br /><br />Capture: <a href="/products/HA723457" style="font-weight: bold;text-decoration: underline;">HA723457</a>, Human CD28 Rabbit mAb [PSH12-50]<br />Detector: <a href="/products/HA723458" style="font-weight: bold;text-decoration: underline;">HA723458</a>, Human CD28 Rabbit mAb [PSH12-51]<br /><br />Interpolated concentration of native CD28 was measured in duplicate at different sample concentrations and interpolated from the CD28 standard curves. The mean CD28 concentration was determined to be 4,345 pg/mL in Jurkat cell extract. There was no detectable signal in THP-1 cell extract.](https://storage.huabio.cn/huabio/productImg/HA723458_3.png?v=20260401002256)
Interpolated concentrations of native CD28 in Jurkat and THP-1 extract samples based on a 1000 µg/ml extract load.
Capture: HA723457, Human CD28 Rabbit mAb [PSH12-50]
Detector: HA723458, Human CD28 Rabbit mAb [PSH12-51]
Interpolated concentration of native CD28 was measured in duplicate at different sample concentrations and interpolated from the CD28 standard curves. The mean CD28 concentration was determined to be 4,345 pg/mL in Jurkat cell extract. There was no detectable signal in THP-1 cell extract. -
![Interpolated concentrations of spiked CD28 in cell culture media samples.<br /><br />Capture: <a href="/products/HA723457" style="font-weight: bold;text-decoration: underline;">HA723457</a>, Human CD28 Rabbit mAb [PSH12-50]<br />Detector: <a href="/products/HA723458" style="font-weight: bold;text-decoration: underline;">HA723458</a>, Human CD28 Rabbit mAb [PSH12-51]<br /><br />The concentrations of CD28 were measured in duplicates, interpolated from the CD28 standard curves and corrected for sample dilution. Undiluted samples are as follows: cell culture media 50%.](https://storage.huabio.cn/huabio/productImg/HA723458_4.png?v=20260401002256)
Interpolated concentrations of spiked CD28 in cell culture media samples.
Capture: HA723457, Human CD28 Rabbit mAb [PSH12-50]
Detector: HA723458, Human CD28 Rabbit mAb [PSH12-51]
The concentrations of CD28 were measured in duplicates, interpolated from the CD28 standard curves and corrected for sample dilution. Undiluted samples are as follows: cell culture media 50%.
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"
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Biotin Conjugated Human CD28 Recombinant Rabbit Monoclonal Antibody [PSH12-51] - Detector
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Reactivity: Human
Conjugate: Biotin
Human CD28 Recombinant Rabbit Monoclonal Antibody [PSH12-50] - BSA and Azide free (Capture)
Application: ELISA(Cap)
Reactivity: Human
Conjugate: unconjugated
