Product nameAnti-CD8 antibody [YTS169.4]
See all CD8 primary antibodies
DescriptionRat monoclonal [YTS169.4] to CD8
Tested applicationsSuitable for: ICC/IF, Flow Cyt, IHC-Frmore details
Species reactivityReacts with: Mouse
CBA mouse spleen cells and thymocytes
- Flow Cyt: Mouse spleen cells.
ab22378 is not expected to work in standard western blots, under denaturing conditions, or in IHC on formalin-fixed tissue. To our knowledge, ab22378 will only stain frozen sections or fresh cells, and will only immunoprecipitate the antigen from cell lysates in mild detergents such as NP40 or Triton-X100.
This antibody clone is manufactured by Abcam.
Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
Storage bufferpH: 7.40
Preservative: 0.02% Sodium azide
Concentration information loading...
- Anti-CD8 antibody [SP16] (ab101500)
- Anti-CD8 antibody [SP239] (ab178089)
- Anti-CD8 antibody [EP1150Y] (Alexa Fluor® 647) (ab196193)
- Anti-CD8 antibody [EP1150Y] (Alexa Fluor® 488) (ab196462)
- Anti-CD8 antibody [EP1150Y] - Low endotoxin, Azide free (ab210067)
- Anti-CD8 antibody [SP239] - BSA and Azide free (ab236230)
- Anti-CD8 antibody [CAL38] (ab237723)
- Anti-CD8 antibody [EP1150Y] (ab93278)
Our Abpromise guarantee covers the use of ab22378 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|ICC/IF||Use at an assay dependent concentration.|
|Flow Cyt||Use 0.1-1µg for 106 cells.
ab18536 - Rat monoclonal IgG2b, is suitable for use as an isotype control with this antibody.
|IHC-Fr||Use at an assay dependent concentration.|
FunctionIdentifies cytotoxic/suppressor T-cells that interact with MHC class I bearing targets. CD8 is thought to play a role in the process of T-cell mediated killing. CD8 alpha chains binds to class I MHC molecules alpha-3 domains.
Involvement in diseaseDefects in CD8A are a cause of familial CD8 deficiency (CD8 deficiency) [MIM:608957]. Familial CD8 deficiency is a novel autosomal recessive immunologic defect characterized by absence of CD8+ cells, leading to recurrent bacterial infections.
Sequence similaritiesContains 1 Ig-like V-type (immunoglobulin-like) domain.
modificationsAll of the five most C-terminal cysteines form inter-chain disulfide bonds in dimers and higher multimers, while the four N-terminal cysteines do not.
Cellular localizationSecreted and Cell membrane.
- Information by UniProt
FormCD8 beta tissue specificity: Isoform 1, isoform 3, isoform 5, isoform 6, isoform 7 and isoform 8 are expressed in both thymus and peripheral CD8+ T-cells. Expression of isoform 1 is higher in thymus CD8+ T-cells than in peripheral CD8+ T-cells. Expression of isoform 6 is higher in peripheral CD8+ T-cells than in thymus CD8+ T-cells. CD8 beta PTM: Phosphorylated as a consequence of T-cell activation.
- CD8 antibody
- CD8 antigen, alpha polypeptide (p32) antibody
- CD8a antibody
IHC-Fr labeling for CD8-positive T-Cells, as indicated by red immunofluorescence (ab22378 at a 1/50 dilution). Yellow arrows designate positive cells. *** = p≤0.001.
Frozen sections from mouse skin (8 µm thick) were post-fixed in cold acetone at −20°C for 10 minutes then air dried at room temperature for 20 minutes. All sections were then washed three times with 0.05% PBS-Tween (PBST); then nonspecific antibody binding was blocked using 10% normal goat serum in PBST at room temperature for 30 minutes. Sections were then incubated overnight at 4°C with primary antibody. Primary antibody incubation was followed by three washes with PBST before addition of the secondary antibodies, which were incubated on the sections for 2 hours at room temperature. Nuclei were counterstained with DAPI (200 ng/mL) for 10 minutes at room temperature. Finally, sections were rinsed with PBST, dehydrated through sequential alcohol washes and then cleared in xylene. Slides were mounted with DPX mounting medium.
Normal mouse (CBA) spleen cells (red cell lysed) double stained with ab22378 (0.3 µg/mL, detected with Alexa Fluor® 488-conjugated monoclonal mouse anti-rat antibody) and an APC conjugated hamster anti-mouse CD3 antibody. ab22378 stains an expected subpopulation of CD8 alpha positive T cells.
Overlay histogram showing mouse spleen cells (C57BL/6) stained with ab22378 (red line). The cells were incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab22378, 0.1μg/1x106 cells) for 30 min at 4ºC. The secondary antibody used was Alexa Fluor® 488 goat anti-rat IgG (H&L) (ab150157) at 1/2000 dilution for 30 min at 4ºC. Isotype control antibody (black line) was rat IgG2b [RTK4530] (ab18541, 2μg/1x106 cells) used under the same conditions.
Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.
ab22378 staining CD8 in murine bone marrow cells by ICC/IF (immunocytochemistry/immunofluorescence).
Cells were formaldehyde fixed prior to blocking in 2% BSA for 30 minutes at 20°C. The primary antibody (1/200) was incubated with the sample for 9 hours at 4°C. An undiluted Alexa Fluor® 555-conjugated goat anti-rat polyclonal was used as the secondary. The nuclei were stained with DAPI.
ab22378 staining CD8 in mouse spleen tissue sections by IHC-Fr (Frozen sections).
Tissue samples were fixed with acetone and blocked with 5% serum for 2 hours at 25°C. The sample was incubated with primary antibody (1/250 in PBS-Tween) at 4°C for 12 hours. An Alexa Fluor® 594-conjugated Goat polyclonal to rat IgG (1/250) was used as secondary antibody. Nuclei were stained with DAPI.
This product has been referenced in:
- Tian L et al. Calotropin regulates the apoptosis of non-small cell cancer by regulating the cytotoxic T-lymphocyte associated antigen 4-mediated TGF-ß/ERK signaling pathway. Mol Med Rep 17:7683-7691 (2018). Read more (PubMed: 29620207) »
- Hsu JM et al. STT3-dependent PD-L1 accumulation on cancer stem cells promotes immune evasion. Nat Commun 9:1908 (2018). Read more (PubMed: 29765039) »