Recombinant Anti-CD8 alpha antibody [CAL66] - BSA and Azide free (ab251596)

Product name

Anti-CD8 alpha antibody [CAL66] - BSA and Azide free
See all CD8 alpha primary antibodies
Description

Rabbit monoclonal [CAL66] to CD8 alpha - BSA and Azide free
Host species

Rabbit
Tested applications

Suitable for: ICC/IF, IP, Flow Cyt, IHC-P, mIHCmore details
Unsuitable for: WB
Species reactivity

Reacts with: Rat, Human
Immunogen

Synthetic peptide within Human CD8 alpha aa 204-235. The exact sequence is proprietary.
Database link: P01732
Positive control
- IHC-P: Human tonsil, colon and gastric carcinoma tissue; rat spleen tissue. ICC/IF: Human PBMCs. Flow cyt: Human PBMCs. IP: Human thymus lysate. mIHC: Human breast cancer tissue.
General notes

ab251596 is the carrier-free version of ab237709.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.

This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
This product is compatible with the Maxpar^® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar^® is a trademark of Fluidigm Canada Inc.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C. Do Not Freeze.
Storage buffer

Constituent: PBS
Carrier free

Yes
Concentration information loading...
Purity

Protein A purified
Purification notes

Purity >99%
Clonality

Monoclonal
Clone number

CAL66
Isotype

IgG
Research areas

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab251596 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.
IP		Use at an assay dependent concentration.
Flow Cyt		Use at an assay dependent concentration.
IHC-P		Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. Use at 1 μg/ml for rat
mIHC		Use at an assay dependent concentration.

Notes
ICC/IF Use at an assay dependent concentration.
IP Use at an assay dependent concentration.
Flow Cyt Use at an assay dependent concentration.
IHC-P Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. Use at 1 μg/ml for rat
mIHC Use at an assay dependent concentration.

Application notes

Is unsuitable for WB.

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.
Target information above from: UniProt accession P01732 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
Database links
- Entrez Gene: 925 Human
- Entrez Gene: 24930 Rat
- Omim: 186910 Human
- SwissProt: P01732 Human
- SwissProt: P07725 Rat
- Unigene: 85258 Human
- Unigene: 10306 Rat
Alternative names
- alpha polypeptide (p32) antibody
- CD_antigen=CD8a antibody
- CD8 antibody
- CD8 antigen alpha polypeptide antibody
- CD8 antigen alpha polypeptide (p32) 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
- Ly 2 antibody
- Ly 35 antibody
- Ly B antibody
- Ly2 antibody
- Ly3 antibody
- Ly35 antibody
- LyB antibody
- Lyt 2.1 lymphocyte differentiation antigen (AA at 100) 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 lymphocyte differentiation antigen T8/Leu 2 antibody
- T-cell surface glycoprotein CD8 alpha chain antibody
- T-cell surface glycoprotein Lyt 2 antibody
- T-lymphocyte differentiation antigen T8/Leu-2 antibody
- T8 T cell antigen antibody
- T8/Leu-2 T-lymphocyte differentiation antigen antibody
see all

Multiplex immunohistochemistry - Anti-CD8 alpha antibody [CAL66] - BSA and Azide free (ab251596)This image is courtesy of ImmunoAtlas.

Fluorescence multiplex immunohistochemical analysis of Human breast cancer tissue (formalin-fixed paraffin-embedded section).

Merged staining of Anti-PD-L1 (ab251611), Anti-Granzyme B (ab219803), Anti-PD1 (ab251613), Anti-pan Cytokeratin (ab264485), Anti-EpCAM (ab225894), Anti-CD8 alpha (ab251596) and Anti-FOXP3 (ab96048).

The immunostaining was performed on a Leica Biosystems BOND^® MAX instrument with an Opal™ 6-Plex Detection Kit (NEL821001KT, Akoya Biosciences^®).

The section was incubated in six rounds of staining; sequentially for ab251611 (1/750 dilution), ab219803 (1/250 dilution), ab251613 (1/750 dilution), ab264485 (0.5 µg/ml), ab225894 (1/1250 dilution), ab251596 (1/1500 dilution) and ab96048 (10 µg/ml); each using a separate fluorescent tyramide signal amplification system. EDTA based antigen retrieval (Leica Biosystems BOND^® Epitope Retrieval Solution 2, pH 9.0, 20 minutes) was used in between rounds of tyramide signal amplification to remove the antibody from the previous round, to avoid any cross-reactivity. DAPI (dark blue) was used as a nuclear counter stain.
Microscopy and pseudocoloring of individual Opal™ dyes was performed using a Vectra 3 Imaging System (Akoya Biosciences^®).

This data was developed using the same antibody clone in a different buffer formulation containing PBS and sodium azide (ab237709).

This data is courtesy of ImmunoAtlas and it can be found here.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD8 alpha antibody [CAL66] - BSA and Azide free (ab251596)

Immunohistochemical analysis of paraffin-embedded human tonsil tissue labeling CD8 alpha with ab237709 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Positive staining on the human tonsil is observed. Counter stained with hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ready to use.

Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

This data was developed using the same antibody clone in a different buffer formulation containing PBS and sodium azide (ab237709).
Multiplex immunohistochemistry - Anti-CD8 alpha antibody [CAL66] - BSA and Azide free (ab251596)This image is courtesy of ImmunoAtlas.

Fluorescence multiplex immunohistochemical analysis of Human breast cancer tissue (formalin-fixed paraffin-embedded section).

Merged staining of Anti-PD-L1 (ab251611), Anti-Granzyme B (ab219803), Anti-PD1 (ab251613), Anti-pan Cytokeratin (ab264485), Anti-EpCAM (ab225894), Anti-CD8 alpha (ab251596) and Anti-FOXP3 (ab96048).

The immunostaining was performed on a Leica Biosystems BOND^® MAX instrument with an Opal™ 6-Plex Detection Kit (NEL821001KT, Akoya Biosciences^®).

The section was incubated in six rounds of staining; sequentially for ab251611 (1/750 dilution), ab219803 (1/250 dilution), ab251613 (1/750 dilution), ab264485 (0.5 µg/ml), ab225894 (1/1250 dilution), ab251596 (1/1500 dilution) and ab96048 (10 µg/ml); each using a separate fluorescent tyramide signal amplification system. EDTA based antigen retrieval (Leica Biosystems BOND^® Epitope Retrieval Solution 2, pH 9.0, 20 minutes) was used in between rounds of tyramide signal amplification to remove the antibody from the previous round, to avoid any cross-reactivity. DAPI (dark blue) was used as a nuclear counter stain.
Microscopy and pseudocoloring of individual Opal™ dyes was performed using a Vectra 3 Imaging System (Akoya Biosciences^®).

This data was developed using the same antibody clone in a different buffer formulation containing PBS and sodium azide (ab237709).

This data is courtesy of ImmunoAtlas and it can be found here.
Multiplex immunohistochemistry - Anti-CD8 alpha antibody [CAL66] - BSA and Azide free (ab251596)This image is courtesy of ImmunoAtlas.

Fluorescence multiplex immunohistochemical analysis of Human breast cancer tissue (formalin-fixed paraffin-embedded section).

Merged staining of Anti-PD-L1 (ab251611), Anti-Granzyme B (ab219803), Anti-PD1 (ab251613), Anti-pan Cytokeratin (ab264485), Anti-EpCAM (ab225894), Anti-CD8 alpha (ab251596) and Anti-FOXP3 (ab96048).

The immunostaining was performed on a Leica Biosystems BOND^® MAX instrument with an Opal™ 6-Plex Detection Kit (NEL821001KT, Akoya Biosciences^®).

The section was incubated in six rounds of staining; sequentially for ab251611 (1/750 dilution), ab219803 (1/250 dilution), ab251613 (1/750 dilution), ab264485 (0.5 µg/ml), ab225894 (1/1250 dilution), ab251596 (1/1500 dilution) and ab96048 (10 µg/ml); each using a separate fluorescent tyramide signal amplification system. EDTA based antigen retrieval (Leica Biosystems BOND^® Epitope Retrieval Solution 2, pH 9.0, 20 minutes) was used in between rounds of tyramide signal amplification to remove the antibody from the previous round, to avoid any cross-reactivity. DAPI (dark blue) was used as a nuclear counter stain.
Microscopy and pseudocoloring of individual Opal™ dyes was performed using a Vectra 3 Imaging System (Akoya Biosciences^®).

This data was developed using the same antibody clone in a different buffer formulation containing PBS and sodium azide (ab237709).

This data is courtesy of ImmunoAtlas and it can be found here.
Immunoprecipitation - Anti-CD8 alpha antibody [CAL66] - BSA and Azide free (ab251596)

CD8 alpha was immunoprecipitated from 0.35 mg of human thymus lysate with ab237709 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab237709 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used as secondary antibody at 1/5000 dilution.

Lane 1: Human thymus lysate 10 μg (Input).
Lane 2: ab237709 IP in human thymus lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab237709 in human thymus lysate.

Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 100 seconds.

This data was developed using the same antibody clone in a different buffer formulation containing PBS and sodium azide (ab237709).
Multiplex immunohistochemistry - Anti-CD8 alpha antibody [CAL66] - BSA and Azide free (ab251596)This image is courtesy of ImmunoAtlas.

Fluorescence multiplex immunohistochemical analysis of Human breast cancer tissue (formalin-fixed paraffin-embedded section).

Merged staining of Anti-PD-L1 (ab251611), Anti-Granzyme B (ab219803), Anti-PD1 (ab251613), Anti-pan Cytokeratin (ab264485), Anti-EpCAM (ab225894), Anti-CD8 alpha (ab251596) and Anti-FOXP3 (ab96048).

The immunostaining was performed on a Leica Biosystems BOND^® MAX instrument with an Opal™ 6-Plex Detection Kit (NEL821001KT, Akoya Biosciences^®).

The section was incubated in six rounds of staining; sequentially for ab251611 (1/750 dilution), ab219803 (1/250 dilution), ab251613 (1/750 dilution), ab264485 (0.5 µg/ml), ab225894 (1/1250 dilution), ab251596 (1/1500 dilution) and ab96048 (10 µg/ml); each using a separate fluorescent tyramide signal amplification system. EDTA based antigen retrieval (Leica Biosystems BOND^® Epitope Retrieval Solution 2, pH 9.0, 20 minutes) was used in between rounds of tyramide signal amplification to remove the antibody from the previous round, to avoid any cross-reactivity. DAPI (dark blue) was used as a nuclear counter stain.
Microscopy and pseudocoloring of individual Opal™ dyes was performed using a Vectra 3 Imaging System (Akoya Biosciences^®).

This data was developed using the same antibody clone in a different buffer formulation containing PBS and sodium azide (ab237709).

This data is courtesy of ImmunoAtlas and it can be found here.
Multiplex immunohistochemistry - Anti-CD8 alpha antibody [CAL66] - BSA and Azide free (ab251596)This image is courtesy of ImmunoAtlas.

Fluorescence multiplex immunohistochemical analysis of Human breast cancer tissue (formalin-fixed paraffin-embedded section).

Merged staining of Anti-PD-L1 (ab251611), Anti-Granzyme B (ab219803), Anti-PD1 (ab251613), Anti-pan Cytokeratin (ab264485), Anti-EpCAM (ab225894), Anti-CD8 alpha (ab251596) and Anti-FOXP3 (ab96048).

The immunostaining was performed on a Leica Biosystems BOND^® MAX instrument with an Opal™ 6-Plex Detection Kit (NEL821001KT, Akoya Biosciences^®).

The section was incubated in six rounds of staining; sequentially for ab251611 (1/750 dilution), ab219803 (1/250 dilution), ab251613 (1/750 dilution), ab264485 (0.5 µg/ml), ab225894 (1/1250 dilution), ab251596 (1/1500 dilution) and ab96048 (10 µg/ml); each using a separate fluorescent tyramide signal amplification system. EDTA based antigen retrieval (Leica Biosystems BOND^® Epitope Retrieval Solution 2, pH 9.0, 20 minutes) was used in between rounds of tyramide signal amplification to remove the antibody from the previous round, to avoid any cross-reactivity. DAPI (dark blue) was used as a nuclear counter stain.
Microscopy and pseudocoloring of individual Opal™ dyes was performed using a Vectra 3 Imaging System (Akoya Biosciences^®).

This data was developed using the same antibody clone in a different buffer formulation containing PBS and sodium azide (ab237709).

This data is courtesy of ImmunoAtlas and it can be found here.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD8 alpha antibody [CAL66] - BSA and Azide free (ab251596)

Immunohistochemical analysis of paraffin-embedded human colon tissue labeling CD8 alpha with ab237709 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Positive staining on the T lymphocytes in human colon is observed. Counter stained with hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ready to use.

Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

This data was developed using the same antibody clone in a different buffer formulation containing PBS and sodium azide (ab237709).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD8 alpha antibody [CAL66] - BSA and Azide free (ab251596)

Immunohistochemical analysis of paraffin-embedded human gastric carcinoma tissue labeling CD8 alpha with ab237709 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Positive staining on the T lymphocytes in human gastric carcinoma is observed. Counter stained with hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ready to use.

Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

This data was developed using the same antibody clone in a different buffer formulation containing PBS and sodium azide (ab237709).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD8 alpha antibody [CAL66] - BSA and Azide free (ab251596)

Immunohistochemical analysis of paraffin-embedded rat spleen tissue labeling CD8 alpha with ab237709 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Positive staining on the rat spleen is observed. Counter stained with hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ready to use.

Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

This data was developed using the same antibody clone in a different buffer formulation containing PBS and sodium azide (ab237709).
Immunocytochemistry/ Immunofluorescence - Anti-CD8 alpha antibody [CAL66] - BSA and Azide free (ab251596)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized human PBMC (peripheral blood mononuclear cells) labeling CD8 alpha with ab237709 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing membranous staining in subsets of human PBMC. The nuclear counter stain is DAPI (blue). CD3 is detected with an anti-CD3 antibody-Alexa Fluor^® 647 comjugate at 1/200 dilution (red).

Secondary antibody only control: Used PBS instead of primary antibody, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS and sodium azide (ab237709).
Flow Cytometry - Anti-CD8 alpha antibody [CAL66] - BSA and Azide free (ab251596)

Flow cytometric analysis of 2% paraformaldehyde-fixed, 0.1% Tween 20 permeacblized human PBMC (peripheral blood mononuclear cells) labeling CD8 alpha with ab237709 at 1/500 dilution (Right) compared with Recombinant Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (Left).

Goat Anti-Rabbit IgG Fc (Alexa Fluor^® 488) preadsorbed (ab150097), at 1/5000 dilution was used as the secondary antibody.

Cells were surface stained with anti-CD8 alpha conjugated to BV421.The fixed with 2% PFA for 10min followed by intracellularly stained with rabbit IgG (Left) or ab237709 (Right). They recognize same populations.

This data was developed using the same antibody clone in a different buffer formulation containing PBS and sodium azide (ab237709).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD8 alpha antibody [CAL66] - BSA and Azide free (ab251596)

Formalin-fixed, paraffin-embedded human tonsil tissue stained for CD8 alpha using ab237709 at 0.25 µg/ml in immunohistochemical analysis.

This data was developed using the same antibody clone in a different buffer formulation containing PBS and sodium azide (ab237709).
Anti-CD8 alpha antibody [CAL66] - BSA and Azide free (ab251596)

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

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Key features and details

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Overview

Product name

Description

Host species

Tested applications

Species reactivity

Immunogen

Positive control

General notes

Properties

Form

Storage instructions

Storage buffer

Carrier free

Purity

Purification notes

Clonality

Clone number

Isotype

Research areas

Associated products

Alternative Versions

Compatible Secondaries

Conjugation kits

Immunohistochemistry reagents

Isotype control

Applications

The Abpromise guarantee

Target

Function

Involvement in disease

Sequence similarities

Post-translationalmodifications

Cellular localization

Database links

Alternative names

Images

Protocols

Datasheets and documents

Datasheet download

References (0)

Customer reviews and Q&As

Post-translational
modifications