Human MICA Recombinant Rabbit Monoclonal Antibody [PSH11-40]
cat.: HA723328
| Product Type: | Recombinant Rabbit monoclonal IgG, primary antibodies |
|---|---|
| Species reactivity: | Human |
| Applications: | ELISA(Det) |
| Clonality: | Monoclonal |
| Clone number: | PSH11-40 |
| Form: | Liquid |
| Storage condition: | Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles. |
| Storage buffer: | PBS (pH7.4). |
| Concentration: | 1ug/ul |
| Purification: | Protein A affinity purified. |
| Isotype: | IgG |
| Immunogen: | Recombinant protein within Human MICA aa 24-308 (HA210951). |
| Positive control: | Recombinant Human MICA protein (HA210951). |
| Subcellular location: | Cell membrane. Cytoplasm. |
| Recommended Dilutions:
ELISA(Det) |
Use at an assay dependent concentration. Can be paired for Sandwich ELISA with Rabbit monoclonal [PSH11-39] to HumanMICA antibody (Capture) (HA723327) and Recombinant Human Human MICA protein (HA210951) as the standard. The reference range value is 31.2-4,000 pg/mL. |
| Uniprot #: | SwissProt: Q29983 Human |
| Alternative names: | HLA class I antigen FLJ36918 FLJ60820 MGC111087 MGC21250 MHC class I chain related gene A protein MHC class I chain related protein A MHC class I chain related protein A HLA B HLA C MHC class I polypeptide related sequence A MHC class I polypeptide-related sequence A MHC class I related protein MIC A MIC-A micA MICA_HUMAN OTTHUMP00000029088 OTTHUMP00000044528 OTTHUMP00000165170 OTTHUMP00000165172 PERB11.1 Stress inducible class I homolog |
Images
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Fig1:
Sandwich ELISA analysis of human MICA matched pair antibodies Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA723327) 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 MICA protein (HA210951) starting from 4,000 pg/ml to 0 pg/ml and detect antibody (HA723328, 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. |
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Fig2:
Interpolated concentrations of native MICA in Hela, K-562 and THP-1 extract samples based on a 1000 µg/ml extract load. Interpolated concentration of native MICA was measured in duplicate at different sample concentrations and interpolated from the MICA standard curves. The interpolated dilution factor corrected values were plotted (mean +/- SD, n=2). The mean MICA concentration was determined to be 7,949 pg/mL in Hela and 1,379 pg/mL in K-562 cell extract. There was no detectable signal in THP-1 cell extract. |
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Fig3:
Interpolated concentrations of spiked MICA in cell culture media samples. The concentrations of MICA were measured in duplicates, interpolated from the MICA 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). |
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Fig4:
ELISA Assay of Anti-Human MICA Antibody (HA723328) to different antigens. Indirect ELISA analysis of Human MICA was performed by coating wells of a 96-well plate with 50 µl per well of Human MICA antigen and Human MICB diluted in carbonate/bicarbonate buffer, at a concentration of 1 µg/mL overnight at 4℃. Wells of the plate were washed, blocked with Starting Block blocking buffer, and incubated with 50 µl per well of a mouse Human MICA monoclonal antibody starting at a concentration of 2 µg/mL (HA723328) and serially diluting it to a concentration of 0.13 ng/mL for 2 hours at room temperature. The plate was washed and incubated with 50 µl per well of an HRP-conjugated goat anti-Rabbit IgG secondary antibody at a dilution of 1:10,000 for one hour at room temperature. Detection was performed using an Ultra TMB Substrate for 5 minutes at room temperature in the dark. The reaction was stopped with sulfuric acid and absorbances were read on a spectrophotometer at 450 nm. |
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.