Name: Anti-CD8 alpha antibody [CAL66]
Brand: RabMAb
SKU: ab237709
Rating: 5 (2 reviews)

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Multiplex immunohistochemistry - Anti-CD8 alpha antibody [CAL66] (ab237709)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [CAL66] to CD8 alpha
Suitable for: ICC/IF, IP, Flow Cyt, mIHC, IHC-P
Reacts with: Rat, Human

Alexa Fluor® 488 Alexa Fluor® 555 Alexa Fluor® 594 Alexa Fluor® 647 Carrier Free

製品名

Anti-CD8 alpha antibody [CAL66]
CD8 alpha 一次抗体製品一覧
製品の詳細

Rabbit monoclonal [CAL66] to CD8 alpha
由来種

Rabbit
アプリケーション

適用あり: ICC/IF, IP, Flow Cyt, mIHC, IHC-Pmore details
適用なし: WB
種交差性

交差種: Rat, Human
免疫原

Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
ポジティブ･コントロール
- 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.

製品の状態

Liquid
保存方法

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
バッファー

pH: 7.2
Preservative: 0.05% Sodium azide
Constituent: PBS
Concentration information loading...
精製度

Protein A purified
特記事項（精製）

Purity >99%
ポリ/モノ

モノクローナル
クローン名

CAL66
アイソタイプ

IgG
研究分野

Alternative Versions
- Anti-CD8 alpha antibody [CAL66] - BSA and Azide free (ab251596)
Compatible Secondaries
Immunohistochemistry reagents
- Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) (ab93684)
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)

The Abpromise guarantee

Abpromise保証は、次のテスト済みアプリケーションにおけるab237709の使用に適用されます

アプリケーションノートには、推奨の開始希釈率がありますが、適切な希釈率につきましてはご検討ください。

アプリケーション	Abreviews	特記事項
ICC/IF		1/100.
IP		1/30.
Flow Cyt		1/500.
mIHC		Use at an assay dependent concentration.
IHC-P	(2)	Use a concentration of 0.25 µg/ml. 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

特記事項
ICC/IF 1/100.
IP 1/30.
Flow Cyt 1/500.
mIHC Use at an assay dependent concentration.
IHC-P Use a concentration of 0.25 µg/ml. 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

追加情報

Is unsuitable for WB.

機能

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.
関連疾患

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.
配列類似性

Contains 1 Ig-like V-type (immunoglobulin-like) domain.
翻訳後修飾

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.
細胞内局在

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
参照データベース
- Entrez Gene: 925 Human
- Entrez Gene: 24930 Rat
- Omim: 186910 Human
- SwissProt: P01732 Human
- SwissProt: P07725 Rat
- Unigene: 85258 Human
- Unigene: 10306 Rat
別名
- 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] (ab237709)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; cyan; Opal™ 520), Anti-Granzyme B (ab219803; yellow; Opal™ 540), Anti-PD1 (ab251613; magenta; Opal™ 570), Anti-pan Cytokeratin (ab264485; red; Opal™ 620), Anti-EpCAM (ab225894; red; Opal™ 620), Anti-CD8 alpha (ab251596; green; Opal™ 650) and Anti-FOXP3 (ab96048; orange; Opal™ 690). EpCAM and pan-cytokeratin share the same dye and color. Dyes are pseudo-colored for better contrast of the markers.

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 is courtesy of ImmunoAtlas and it can be found here.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD8 alpha antibody [CAL66] (ab237709)

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).
Multiplex immunohistochemistry - Anti-CD8 alpha antibody [CAL66] (ab237709)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; cyan; Opal™ 520), Anti-Granzyme B (ab219803; yellow; Opal™ 540), Anti-PD1 (ab251613; magenta; Opal™ 570), Anti-pan Cytokeratin (ab264485; red; Opal™ 620), Anti-EpCAM (ab225894; red; Opal™ 620), Anti-CD8 alpha (ab251596; green; Opal™ 650) and Anti-FOXP3 (ab96048; orange; Opal™ 690). EpCAM and pan-cytokeratin share the same dye and color. Dyes are pseudo-colored for better contrast of the markers.

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 is courtesy of ImmunoAtlas and it can be found here.
Multiplex immunohistochemistry - Anti-CD8 alpha antibody [CAL66] (ab237709)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; cyan; Opal™ 520), Anti-Granzyme B (ab219803; yellow; Opal™ 540), Anti-PD1 (ab251613; magenta; Opal™ 570), Anti-pan Cytokeratin (ab264485; red; Opal™ 620), Anti-EpCAM (ab225894; red; Opal™ 620), Anti-CD8 alpha (ab251596; green; Opal™ 650) and Anti-FOXP3 (ab96048; orange; Opal™ 690). EpCAM and pan-cytokeratin share the same dye and color. Dyes are pseudo-colored for better contrast of the markers.

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 is courtesy of ImmunoAtlas and it can be found here.
Immunoprecipitation - Anti-CD8 alpha antibody [CAL66] (ab237709)

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.
Multiplex immunohistochemistry - Anti-CD8 alpha antibody [CAL66] (ab237709)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; cyan; Opal™ 520), Anti-Granzyme B (ab219803; yellow; Opal™ 540), Anti-PD1 (ab251613; magenta; Opal™ 570), Anti-pan Cytokeratin (ab264485; red; Opal™ 620), Anti-EpCAM (ab225894; red; Opal™ 620), Anti-CD8 alpha (ab251596; green; Opal™ 650) and Anti-FOXP3 (ab96048; orange; Opal™ 690). EpCAM and pan-cytokeratin share the same dye and color. Dyes are pseudo-colored for better contrast of the markers.

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 is courtesy of ImmunoAtlas and it can be found here.
Multiplex immunohistochemistry - Anti-CD8 alpha antibody [CAL66] (ab237709)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; cyan; Opal™ 520), Anti-Granzyme B (ab219803; yellow; Opal™ 540), Anti-PD1 (ab251613; magenta; Opal™ 570), Anti-pan Cytokeratin (ab264485; red; Opal™ 620), Anti-EpCAM (ab225894; red; Opal™ 620), Anti-CD8 alpha (ab251596; green; Opal™ 650) and Anti-FOXP3 (ab96048; orange; Opal™ 690). EpCAM and pan-cytokeratin share the same dye and color. Dyes are pseudo-colored for better contrast of the markers.

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 is courtesy of ImmunoAtlas and it can be found here.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD8 alpha antibody [CAL66] (ab237709)

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).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD8 alpha antibody [CAL66] (ab237709)

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).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD8 alpha antibody [CAL66] (ab237709)

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).
Immunocytochemistry/ Immunofluorescence - Anti-CD8 alpha antibody [CAL66] (ab237709)

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.
Flow Cytometry - Anti-CD8 alpha antibody [CAL66] (ab237709)

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 a 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.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD8 alpha antibody [CAL66] (ab237709)

Formalin-fixed, paraffin-embedded human tonsil tissue stained for CD8 alpha using ab237709 at 0.25 µg/ml in immunohistochemical analysis.
Anti-CD8 alpha antibody [CAL66] (ab237709)

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

SDS download

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ab237709 を使用した論文を発表された方は、こちらまでお知らせください。データシートに掲載させていただきます。

ab237709 は 43 報の論文で使用されています。

Qiao T et al. Combined pembrolizumab and bevacizumab therapy effectively inhibits non-small-cell lung cancer growth and prevents postoperative recurrence and metastasis in humanized mouse model. Cancer Immunol Immunother 72:1169-1181 (2023). PubMed: 36357599
Cai Z et al. BCAT2 Shapes a Noninflamed Tumor Microenvironment and Induces Resistance to Anti-PD-1/PD-L1 Immunotherapy by Negatively Regulating Proinflammatory Chemokines and Anticancer Immunity. Adv Sci (Weinh) 10:e2207155 (2023). PubMed: 36642843
Cui B et al. Dendritic cells originating exosomal miR-193b-3p induces regulatory T cells to alleviate liver transplant rejection. Int Immunopharmacol 114:109541 (2023). PubMed: 36700764
Gao L et al. Identification and validation of pyroptosis-related gene landscape in prognosis and immunotherapy of ovarian cancer. J Ovarian Res 16:27 (2023). PubMed: 36707884
Zhao X et al. Adult spinal cord tissue transplantation combined with local tacrolimus sustained-release collagen hydrogel promotes complete spinal cord injury repair. Cell Prolif 56:e13451 (2023). PubMed: 36916024

View all Publications for this product

レビューと Q&A

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Application

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)

Sample

Dog Tissue sections (lymph node)

Antigen retrieval step

Heat mediated - Buffer/Enzyme Used: buffer pH9

Permeabilization

Specification

lymph node

Blocking step

Serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 25°C

Fixative

Formaldehyde

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

投稿 Nov 10 2021

Application

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)

Sample

Monkey Tissue sections (Skin)

Antigen retrieval step

Heat mediated - Buffer/Enzyme Used: buffer pH9

Permeabilization

Specification

Skin

Blocking step

Serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 25°C

Fixative

Formaldehyde

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

投稿 Nov 10 2021

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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Key features and details

Related conjugates and formulations

製品の概要

製品名

製品の詳細

由来種

アプリケーション

種交差性

免疫原

ポジティブ･コントロール

製品の特性

製品の状態

保存方法

バッファー

精製度

特記事項（精製）

ポリ/モノ

クローン名

アイソタイプ

研究分野

関連製品

Alternative Versions

Compatible Secondaries

Immunohistochemistry reagents

Isotype control

アプリケーション

The Abpromise guarantee

ターゲット情報

機能

関連疾患

配列類似性

翻訳後修飾

細胞内局在

参照データベース

別名

画像

プロトコール

データシートおよび資料

SDS download

Datasheet download

参考文献 (43)

レビューと Q&A

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-CD8 alpha antibody [CAL66]

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-CD8 alpha antibody [CAL66]