Overview

  • Product name
    Anti-CD8 antibody [YTS169.4]
    See all CD8 primary antibodies
  • Description
    Rat monoclonal [YTS169.4] to CD8
  • Host species
    Rat
  • Tested applications
    Suitable for: ICC/IF, Flow Cyt, IHC-Frmore details
  • Species reactivity
    Reacts with: Mouse
  • Immunogen

    CBA mouse spleen cells and thymocytes

  • Positive control
    • Flow Cytometry: Mouse Spleen Cells
  • General notes

    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.

    If you require this antibody in a particular buffer formulation or a particular conjugate for your experiments, please contact orders@abcam.com or you can find further information here.

Properties

Applications

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.

Application Abreviews Notes
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.

Target

  • Function
    Identifies 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 disease
    Defects 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 similarities
    Contains 1 Ig-like V-type (immunoglobulin-like) domain.
  • Post-translational
    modifications
    All 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 localization
    Secreted and Cell membrane.
  • Information by UniProt
  • Database links
  • Form
    CD8 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.
  • Alternative names
    • CD8 antibody
    • CD8 antigen, alpha polypeptide (p32) antibody
    • CD8a antibody
    • CD8a antigen antibody
    • CD8a molecule antibody
    • CD8A_HUMAN antibody
    • Leu2 antibody
    • Leu2 T lymphocyte antigen antibody
    • MAL antibody
    • OKT8 T cell antigen antibody
    • p32 antibody
    • T cell antigen Leu2 antibody
    • T cell co receptor antibody
    • T-cell surface glycoprotein CD8 alpha chain antibody
    • T-lymphocyte differentiation antigen T8/Leu-2 antibody
    • T8 T cell antigen antibody
    see all

Images

  • 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.

    See Abreview

  • 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.

    See Abreview

  • 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/1x10^6 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/1x10^6 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.

References

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) »
See all 30 Publications for this product

Customer reviews and Q&As

1-10 of 20 Abreviews or Q&A

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (tumor tissue)
Permeabilization
Yes - 5% Triton X-100 in PBS
Specification
tumor tissue
Blocking step
Serum as blocking agent for 30 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Dec 27 2016

Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (MC38 tumor)
Permeabilization
Yes - 1x PBS with 0.3% triton-x-100
Specification
MC38 tumor
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C
Fixative
Formaldehyde

Dr. Ravikumar Muthuswamy

Verified customer

Submitted May 11 2015

Application
Immunohistochemistry (Frozen sections)
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 24°C
Sample
Mouse Tissue sections (skin wound)
Specification
skin wound
Permeabilization
No
Fixative
Acetone

Abcam user community

Verified customer

Submitted Mar 28 2014

Answer

Thank you for your email.

There may be some antibodies where I might have suggested the human tissues or cells. I apologize for this. I have included the positive control information irrespective of species so similar mouse tissues sections can used as positive control.

Actually the positive control information for each antibody is present on the datasheet; please check the datasheet. Alternatively, we always follow human protein Atlas database (http://www.proteinatlas.org/index.php). The second option is, searching positive control for other antibodies in catalogue against same target; the cell line or tissue sections tested with those antibodies could also be used as positive control. The third place to search for is literature.

I hope this information will be helpful. Should you have any question please do not hesitate to contact me.

Read More

Answer

Thank you for contacting us.

The positive control for ab36392 is IL2 activatedcultured NK cells.

ab9918; human tonsil sections, human cervical squamous cell carcinoma section

ab34674; human artery or colon tissue sections

ab15537; HepG2 cells. mouse brain sections, human placenta sections; Human hepatocellular liver carcinoma cell line; Human hepatocarcinoma tissue sections

ab2557; mouse spleen tissue sections

ab9475; human tonsil tissue sections

ab22378; mouse spleen tissue sections

ab51312; Thymus tissue sections, human pancreas tissue sections.

The best database to search for positive control is Human Protein Atlas. Please click the link http://www.proteinatlas.org/index.php

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Use our products? Submit an Abreview. Earn rewards!
https://www.abcam.com/abreviews

Read More

Answer

Thank you for contacting us.

We suggest to optimise the working concentration according to the sample. Initially, you could try using 1:250 for ab22378 and 2ug/ml for ab119857.

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Use our products? Submit an Abreview. Earn rewards!
https://www.abcam.com/abreviews

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (Lymph Node)
Specification
Lymph Node
Fixative
Acetone/Alcohol
Permeabilization
No
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 23°C

Abcam user community

Verified customer

Submitted Mar 19 2012

Answer

Thank you for your query.
Unfortunately we have not mapped this clone and we currently do not have any empirical data to indicate which region of CD8 is recognized.
However, online papers mentioning the clone indicate it recognizes a band of about 32kDa in lysates. This would suggest that it recognizes CD8b but we have no paper conclusively stating such.
We also have not tested this antibody in WB and it is therefore not covered by our guarantee.
I am sorry that I do not have a more conclusive answer for your question on this occasion and hope it is nevertheless helpful. Please do not hesitate to contact me again with any further questions.

Read More

Answer

Thank you for your inquiry ab85792. Unfortunately, we are not able to release the immunogen sequence for this particular antibody as it is proprietary information. We aim to provide as much information as possible to our customers, so I am sorry that this has not been possible on this occasion. However, I have done a blast search and due to sequence homology ab85792 is expected to cross-react with two species with Pongo pygmaeus and Gorilla gorilla gorilla - but not with mouse. We have two alternative antibodies in our catalogue which cross-react with mouse CD8 i.e ab22378 and ab34282. I would suggest that your customers should take a look at the datasheets to see if they suit their need. Should you have any further questions, please do not hesitate to contact us.

Read More

Answer

Thank you for contacting us. Your credit note ID is XXXX. I am sorry that this antibody did not perform as stated on the datasheet, I have asked our Finance department to issue a credit note for you. The credit note may be used in one of the following ways: (1) Redeemed against the original invoice if this hasn't already been paid. (2) Held on the account for use against a future order. (3) A full refund can be offered where no other invoices are outstanding. Please contact your Finance department to confirm how you would like the credit note to be used and ensure it is not redeemed without your knowledge. To specifically receive a refund please ask your Finance department to contact our Finance department at creditcontrol@abcam.com or by telephone using the information at the “Contact Us” link in the top right corner of our website. The credit note ID is for your reference only, please refer to the credit note ID in any correspondence with our accounting department. We will send you the completed credit note by email or postal mail with the actual credit note number which will start with the letters CGB. I hope this experience will not prevent you from purchasing other products from us in the future. Our Scientific Support team is always at your service should you require further expert advice

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1-10 of 20 Abreviews or Q&A

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