CD8B Antibody (OASA02440)
This product is Genway GWB-3E5851
|Predicted Species Reactivity||Human|
|Product Format||Purified IgG - liquid|
|Additional Information||Fusion Partners: Spleen cells from immunised BALB/c mice were fused with cells of the mouse NS1 myeloma cell line.|
|::||Preservative Stabilisers: 0.09% - Sodium Azide|
0.5% - Bovine Serum Albumin
|::||Approx Protein Conc: IgG concentration 1.0 mg/ml|
Buffer Solutions: Phosphate buffered saline pH7.4
|Reconstitution and Storage||Store at +4oC or at -20oC if preferred.|
This product should be stored undiluted.
Storage in frost free freezers is not recommended. Avoid repeated freezing and thawing as this may denature the antibody. Should this product contain a precipitate we recommend microcentrifugation before use.
|Replacement Item||This antibody may replace item sc-1143 from Santa Cruz Biotechnology.|
|Immunogen||The immunogen for anti-CD8B antibody: normal human T lymphocytes|
|Predicted Homology Based on Immunogen Sequence||Human|
|Datasheets/Manuals||Printable datasheet for anti-CD8B antibody - OASA02440|
|Application Info||Flow Cytometry: Neat - 1/10|
|Protocol Information||Citation: 1: Dennis A, Kudo T, Kruidenier L, Girard F, Crepin VF, MacDonald TT, Frankel G, Wiles S. The p50 subunit of NF-kappaB is critical for in vivo clearance of thenoninvasive enteric pathogen Citrobacter rodentium. Infect Immun. 2008Nov;76(11):4978-88. Epub 2008 Aug 11. PubMed PMID: 18694964; PubMed CentralPMCID: PMC2573329.|
Species: The bacterial strain Citrobacter rodentium, Mouse
Experiment Name: 1. Immunohistochemistry and 2. Determination of C. rodentium-specific antibody responses.
Experiment Background: 1. In this work, Alison et al. characterized the immune response and pathology of mice lacking the p50 subunit of the transcription factor nuclear factor kappa B (NF-ĸB) during C. rodentiuminfection. They showed that p50–/– mice are unable to clear C. rodentium infection. 2. Mice lacking p50 show delayed and reduced immune cell infiltration into colonic tissue during C. rodentium infection. Immune cell infiltrates were measured during C. rodentium infection of p50+/+ mice and p50–/– mice using frozen sections of murine colon incubated with antibodies against CD3 , CD4, CD8, CD11c as a marker for dendritic cells [DC], and F4/80 as a marker for macrophages [Mø]
Experimental Steps: 1 Immunohistochemistry: Cryostat sections of murine colon were rehydrated in Tris-buffered saline (TBS) for 5 min and then incubated with antibodies against CD3, CD4, CD8, F4/80, CD11c, NK1.1, or CD45R (Oxford, United Kingdom) at a 1:10 to 1:100 dilution for 1 h. The sections were gently washed three times with TBS before the addition of biotinylated anti-rat or anti-hamster immunoglobulin G (IgG) , as appropriate, at a dilution of 1:200 with 4% (vol/vol) normal murine serum for blocking (Sera Laboratories International,Horsted Keynes, United Kingdom) for 30 min. After the washing, a 1:200 dilution of 0.1% avidin-peroxidase (Sigma-Aldrich) was added for 30 min before further washing and the addition of diaminobenzadine substrate (Sigma-Aldrich) for 5 to 10 min. The reaction was stopped with excess TBS, and the sections were counterstained with hematoxylin and mounted. A control slide using no primary antibody was also used to show endogenous peroxidase-containing cells. Stained cell populations were counted in five randomly selected fields per section, and the data were expressed as the number of T cells per 250 um2 of lamina propria.2 Determination of C. rodentium-specific antibody responses. An 18-h culture of C. rodentium was resuspended in PBS plus 1% bacterial protease inhibitor (Sigma-Aldrich Ltd., Dorset, United Kingdom) to an optical density at 600 nm (OD600) of 1.0 and then heat killed at 60C for 1 h. A 1:50 PBS dilution of this culture was added to ELISA plates (BD Biosciences, Oxford, United Kingdom) at 100 ul per well before incubation at 4C overnight. The plates were washed three times with 200 ul per well PBS plus 0.05% Tween and then blocked for 1 h with 200 ul per well 1% bovine serum albumin in PBS blocking buffer. At selected time points postinoculation, blood was collected from the mice by cardiac puncture and centrifuged at 8,000 x g to separate red blood cells from serum. The C. rodentium-coated ELISA plates were washed. and then serum samples diluted 1:100 with PBS were added in duplicate wells (50 ul per well) and incubated at room temperature for 2 h. The plates were washed as before and then incubated at room temperature for 2 h with 100 ul per well 1:2,000 blocking buffer dilutions of rat anti-mouse IgG, IgG2b, IgG3, or IgM conjugated to alkaline phosphatase (AbD , Oxford, United Kingdom). After the plates were washed, antibodies were detected with 100 ul per well 1-mg/ml p-nitrophenylphosphate (Sigma-Aldrich Ltd., Dorset, United Kingdom) for 5 to 10 min before the reaction was stopped with 25 ul per well 3 N NaOH. Absorbance was determined using a VersaMax microplate reader with SoftmaxPRO software (Molecular Devices,California) at 405 nm.
Other Reagents Used: kanamycin , nalidixic acid
Number Of Protocols: 2
|Target Reference||1. Terry, L.A. et al. (1990) Differential expression and regulation of the human CD8 alpha and CD8 beta chains. Tissue Antigens 35: 82-91.|
2. DiSanto, J.P. et al. (1991) Generation of anti-human CD8 beta specific antibodies using transfectants expressing mixed species CD8 heterodimers. J. Immunol. Methods 141: 123-131.
3. Yoshino, N. et al. (2000) Upgrading of flow cytometric analysis for absolute counts, cytokines and other antigenic molecules of Cynomolgus monkeys (Macaca fascicularis) by using anti-human cross-reactive antibodies. Exp. Anim. 49: 97-110.
|Alias Symbols||LY3, P37, LEU2, LYT3, CD8B1, MGC119115, CD8B|
|NCBI Gene Id||926|
|Protein Name||T-cell surface glycoprotein CD8 beta chain|
|Description of Target||MOUSE ANTI HUMAN CD8 BETA|
|Protein Accession #||NP_004922.1|
|Protein Size (# AA)||210|
|Tissue Tool||Find tissues and cell lines supported by DNA array analysis to express CD8B.|
|RNA Seq||Find tissues and cell lines supported by RNA-seq analysis to express CD8B.|
- Reconstitution & Storage Instructions
- Western Blotting/Immunoblotting (WB/IB) Protocol
- Immunohistochemistry (IHC) Protocol
- Immunocytochemistry (ICC) Protocol
- Enzyme-Linked ImmunoSorbent Assay (ELISA) Protocol
- Blocking Peptide Competition Protocol (BPCP)
- Immunoprecipitation (IP) Protocol
- Antibody Array (AA) Protocol
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