Overview

  • Product name
    Anti-CD8 alpha antibody [144B]
    See all CD8 alpha primary antibodies
  • Description
    Mouse monoclonal [144B] to CD8 alpha
  • Host species
    Mouse
  • Tested applications
    Suitable for: IHC-R, WB, Flow Cyt, IHC-Pmore details
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Synthetic peptide corresponding to Human CD8 alpha (C terminal).
    Database link: P01732

  • Positive control
    • IHC-P: Human tonsil tissue. Flow Cyt: Human peripheral blood lymphocytes.

Properties

Applications

Our Abpromise guarantee covers the use of ab17147 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
IHC-R Use at an assay dependent concentration.
WB Use at an assay dependent concentration.
Flow Cyt 1/100.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

IHC-P 1/25 - 1/50. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

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 carboxyl-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
  • Alternative names
    • alpha polypeptide (p32) antibody
    • CD8 antibody
    • CD8 antigen alpha polypeptide 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
    • Ly3 antibody
    • LYT3 antibody
    • MAL antibody
    • OKT8 T cell antigen antibody
    • OTTHUMP00000160760 antibody
    • OTTHUMP00000160764 antibody
    • OTTHUMP00000203528 antibody
    • OTTHUMP00000203721 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
    • T8/Leu-2 T-lymphocyte differentiation antigen antibody
    see all

Images

  • Formalin fixed paraffin embedded human tonsil stained with CD8 using ABC and DAB chromogen.
  • Human peripheral blood lymphocytes stained with ab17147 (red line). Human whole blood was processed using a modified protocol based on Chow et al, 2005 (PMID: 16080188). In brief, human whole blood was fixed in 4% formaldehyde (methanol-free) for 10 min at 22°C. Red blood cells were then lyzed by the addition of Triton X-100 (final concentration - 0.1%) for 15 min at 37°C. For experimentation, cells were treated with 50% methanol (-20°C) for 15 min at 4°C. Cells were then incubated with the antibody (ab17147, 1/100 dilution) for 30 min at 4°C. The secondary antibody used was Alexa Fluor® 488 goat anti-mouse IgG (H&L) (ab150113) at 1/2000 dilution for 30 min at 4°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >30,000 total events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter. Gating strategy - peripheral blood lymphocytes.

  • ab17147 staining CD8 in human nasal polyp and tonsil tissue sections by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections). Tissue was fixed with 10% formalin for 24 hours and a heat mediated antigen retrieval step was performed using EDTA pH 8.0. Samples were then blocked with 20% DMEM and 1% BSA in culture medium for 20 minutes at 21°C followed by incubation with the primary antibody at a 1/150 dilution, for 18 hours at 4°C. A biotinylated rabbit anti-mouse (Fab2) polyclonal was used as secondary antibody at a 1/400 dilution.An Avidin Biotin blocking step was used in addition to those already mentioned.

    See Abreview

  • All lanes : Anti-CD8 alpha antibody [144B] (ab17147) at 1/500 dilution

    Lane 1 : Peripheral T-lymphocyte lysate
    Lane 2 : PC3 whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : AP-conjugated polyclonal goat anti-mouse IgG at 1/2000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Observed band size: 29 kDa
    why is the actual band size different from the predicted?


    Exposure time: 15 minutes


    Blocked with 4% BSA for 2 hours at 20°C.

    Incubated with the primary antibody for 30 minutes.

    See Abreview

References

This product has been referenced in:
  • Liu B  et al. PD-1/PD-L1 expression in a series of intracranial germinoma and its association with Foxp3+ and CD8+ infiltrating lymphocytes. PLoS One 13:e0194594 (2018). Read more (PubMed: 29617441) »
  • Fang J  et al. Prognostic significance of tumor infiltrating immune cells in oral squamous cell carcinoma. BMC Cancer 17:375 (2017). Read more (PubMed: 28549420) »
See all 10 Publications for this product

Customer reviews and Q&As

1-8 of 8 Abreviews or Q&A

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (spleen)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Tris/EDTA pH 9.0
Permeabilization
No
Specification
spleen
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 20% · Temperature: RT°C
Fixative
10% NBF

Abcam user community

Verified customer

Submitted Jul 11 2017

Answer

Thank you for contacting us.

We currently do not offer products which meet each of the requirements that you have set. I assembled for you well characterized products which meet most of these requirements but each has some deficiency. Pricing is to be found near the top of the webpage for each product directly below the bold type face product name. There is not currently any special pricing at JHMI. Each datasheet contains all images which we currently have for the products. Any publications which we are aware of the product being used in are located behind the specific references tab or at the bottom of the page.

I would recommend rabbit antibodies as it appears you want to test in mouse however I have added mouse antibodies should you wish to use a mouse on mouse IHC kit such as theMouse on Mouse (M.O.M) Polymer IHC Kit ab127055:https://www.abcam.com/Mouse-on-Mouse-MOM-Polymer-IHC-Kit-ab127055.html



Unconjugated anti-CD3 that targets rat, mouse and pig produced in any animal:

ab16669; rabbit monoclonal; predicted based on sequence homology but not yet tested in Pig

https://www.abcam.com/CD3-antibody-SP7-ab16669.html

Or

ab8671; Mouse monoclonal; predicted but not tested in Pig based on sequence homology; not yet tested in IHC-P

https://www.abcam.com/CD3-antibody-B355-1-RIV-9-ab8671.html



Unconjugated anti-CD4 that targets rat, mouse and pig produced in that same animal:

Ab61131; Rabbit polyclonal; predicted but not tested in Pig, Mouse, Rat based on sequence homology

https://www.abcam.com/CD4-antibody-ab61131.html

Or

ab125711; Mouse monoclonal;predicted in pig; not yet tested in mouse

https://www.abcam.com/CD4-antibody-10B5-ab125711.html



Unconjugated anti-CD8 that targets rat, mouse and pig produced in that same animal:

Ab4055; Rabbit polyclonal, predicted but not tested in Pig, Mouse, Rat based on sequence homology

https://www.abcam.com/CD8-antibody-ab4055.html

Or

Ab17147; Mouse monoclonal, not yet tested in Mouse, Rat or Pig. I do not that the exact immunogen however the C-terminal end is well conserved in each species as such we predict this product may work in these species

https://www.abcam.com/CD8-antibody-144B-ab17147.html



Unconjugated anti-FOXP3 that targets rat, mouse and pig produced in that same animal:

ab75763; Rabbit polyclonal; not yet tested in rat and pig in IHC-P This product is the only rabbit host which is predicted to work in all these species;

https://www.abcam.com/FOXP3-antibody-ab75763.html

Or

ab22510; Mouse monoclonal; predicted but not yet tested in mouse and pig based on sequence homology

https://www.abcam.com/FOXP3-antibody-mAbcam-22510-ab22510.html



Horseradish peroxidase-conjugated anti-that same animal IgG:

Ab97051; Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (HRP)

https://www.abcam.com/Goat-polyclonal-Secondary-Antibody-to-Rabbit-IgG-H-L-HRP-ab97051.html

Or

Ab97023; Goat polyclonal Secondary Antibody to Mouse IgG - H&L (HRP)

https://www.abcam.com/Goat-polyclonal-Secondary-Antibody-to-Mouse-IgG-HL-HRP-ab97023.html



For UNTESTED species and/or applications, we have established a testing discount program. Here is a brief description of how it works:

The testing discount program is for customers who like to use an antibody/protein on an untested species/application. You would purchase the antibody at full price, test it and submit an Abreview with your data (positive or negative). On your next order you will receive a discount for ONE antibody at the full price (100%) of the antibody you have tested. The terms and conditions applicable to this offer can be found here: https://www.abcam.com/index.html?pageconfig=resource&rid=11998&viapagetrap=collaborationdiscount

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 kindly confirming these details. I am sorry this vial of antibody has not worked for this customer.

As requested, we are pleased to arrange a free of charge replacement. I can confirm that 1 vial of ab17147 Anti-CD8 antibody [144B]and ab123946 Anti-Macrophage Scavenger Receptor I antibodyhave been added to your Abcam order reference 1111111. It wasn't possible to send this replacements out with your Friday order, as this will be packed before we Scientific Supporters working. I am sorry about this.

I would like to reassure you that this free of charge replacement vial is also covered by our Abpromise guarantee. Should the customer still be experiencing difficulties with the new vial, or if you have any further questions, please do not hesitate to let me know.

Thank you for your help and cooperation with this case. Please do not hesitate to contact me if you need anything further.

Read More

Answer

Thank you for contacting us. While we have not tested this product in rabbit samples, BLASTs of the immunogen region show that the immunogen used has only a 67% (CD8A) and 46% (CD8B) homology with rabbit. We consider a baseline homology of 85% necessary to ensure cross reactivity and would therefore not recommend this product in rabbit tissues. I believe that you have also contacted us with a further list of products. My colleague is presently gathering information regarding possible alternatives for this and the other targets which you have inquired about and should be in contact with you soon. 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 (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (Nasal Polyp and Tonsil)
Specification
Nasal Polyp and Tonsil
Fixative
Formaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: EDTA, pH8
Permeabilization
No
Blocking step
20% DMEM and 1% BSA in culture medium as blocking agent for 20 minute(s) · Concentration: 1% · Temperature: 21°C

Abcam user community

Verified customer

Submitted Sep 28 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Resin sections)
Sample
Human Tissue sections (Nasal polyp and tonsil)
Specification
Nasal polyp and tonsil

Abcam user community

Verified customer

Submitted Aug 24 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (skin - psoriasis)
Specification
skin - psoriasis
Fixative
Paraformaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: citrate pH 6.0
Permeabilization
Yes - TBST
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 2% · Temperature: 22°C

Miss. Sandra Peternel

Verified customer

Submitted Apr 08 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (Peripheral T-lymphoyctes, PC3 cells)
Loading amount
20 µg
Specification
Peripheral T-lymphoyctes, PC3 cells
Gel Running Conditions
Reduced Denaturing
Blocking step
BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 4% · Temperature: 20°C

Abcam user community

Verified customer

Submitted Jan 15 2008

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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