Name: Anti-CD45 antibody [EP322Y]
Brand: RabMAb
SKU: ab40763
Rating: 5 (8 reviews)

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データシート
文献 (12)
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製品名

Anti-CD45 antibody [EP322Y]
CD45 一次抗体製品一覧
製品の詳細

Rabbit monoclonal [EP322Y] to CD45
由来種

Rabbit
特異性

ab40763 recognizes cytoplasmic domain of CD45.
アプリケーション

適用あり: WB, Flow Cyt, ICC/IF, IHC-P, IHC-Frmore details
種交差性

交差種: Human
免疫原

Synthetic peptide corresponding to residues in the C-terminal domain of human CD45.
ポジティブ･コントロール
- Flow Cyt : Jurkat cells. IHC-P: Human kidney, colon lymphoid, tonsil and spleen tissue. ICC/IF: Jurkat cells. WB: Jurkat, HuT-78 and Raji cell lysate.
特記事項

Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab^® patents
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.
This product is a recombinant rabbit monoclonal antibody.

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. Avoid freeze / thaw cycle.
解離定数（K_D 値）

K_D = 3.60 x 10 ^-11 M
Learn more about K_D
バッファー

pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 40% Glycerol, 0.05% BSA, PBS
Concentration information loading...
精製度

Protein A purified
ポリ/モノ

モノクローナル
クローン名

EP322Y
アイソタイプ

IgG
研究分野

Our Abpromise guarantee covers the use of ab40763 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

アプリケーション	Abreviews	特記事項
WB		1/5000. Detects a band of approximately 200 kDa (predicted molecular weight: 147 kDa).
Flow Cyt		1/15 - 1/20. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. Permeabilization and intracellular staining is necessary.
ICC/IF		1/100.
IHC-P		1/250.
IHC-Fr		Use at an assay dependent concentration.

機能

Protein tyrosine-protein phosphatase required for T-cell activation through the antigen receptor. Acts as a positive regulator of T-cell coactivation upon binding to DPP4. The first PTPase domain has enzymatic activity, while the second one seems to affect the substrate specificity of the first one. Upon T-cell activation, recruits and dephosphorylates SKAP1 and FYN.
関連疾患

Defects in PTPRC are a cause of severe combined immunodeficiency autosomal recessive T-cell-negative/B-cell-positive/NK-cell-positive (T(-)B(+)NK(+) SCID) [MIM:608971]. A form of severe combined immunodeficiency (SCID), a genetically and clinically heterogeneous group of rare congenital disorders characterized by impairment of both humoral and cell-mediated immunity, leukopenia, and low or absent antibody levels. Patients present in infancy recurrent, persistent infections by opportunistic organisms. The common characteristic of all types of SCID is absence of T-cell-mediated cellular immunity due to a defect in T-cell development.
Genetic variations in PTPRC are involved in multiple sclerosis susceptibility (MS) [MIM:126200]. MS is a neurodegenerative disorder characterized by the gradual accumulation of focal plaques of demyelination particularly in the periventricular areas of the brain. Peripheral nerves are not affected. Onset usually in third or fourth decade with intermittent progression over an extended period. The cause is still uncertain.
配列類似性

Belongs to the protein-tyrosine phosphatase family. Receptor class 1/6 subfamily.
Contains 2 fibronectin type-III domains.
Contains 2 tyrosine-protein phosphatase domains.
ドメイン

The first PTPase domain interacts with SKAP1.
翻訳後修飾

Heavily N- and O-glycosylated.
細胞内局在

Membrane. Membrane raft. Colocalized with DPP4 in membrane rafts.
Target information above from: UniProt accession P08575 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: 5788 Human
- Omim: 151460 Human
- SwissProt: P08575 Human
- Unigene: 654514 Human
別名
- B220 antibody
- CD 45 antibody
- CD45 antibody
- CD45 antigen antibody
- CD45R antibody
- GP180 antibody
- L-CA antibody
- LCA antibody
- Leukocyte common antigen antibody
- loc antibody
- Ly-5 antibody
- LY5 antibody
- Ly5, homolog of antibody
- Lyt-4 antibody
- OTTHUMP00000033813 antibody
- OTTHUMP00000033816 antibody
- OTTHUMP00000033817 antibody
- OTTHUMP00000038574 antibody
- Protein tyrosine phosphatase receptor type c polypeptide antibody
- Protein tyrosine phosphatase, receptor type C antibody
- protein tyrosine phosphatase, receptor type, C antibody
- Protein tyrosine phosphatase, receptor type, c polypeptide antibody
- Ptprc antibody
- PTPRC_HUMAN antibody
- Receptor-type tyrosine-protein phosphatase C antibody
- T200 antibody
- T200 glycoprotein antibody
- T200 leukocyte common antigen antibody
see all

Flow Cytometry - Anti-CD45 antibody [EP322Y] (ab40763)

Overlay histogram showing Jurkat (Human T cell leukemia cell line from peripheral blood) cells fixed in 4% PFA and stained with purified ab40763 at a dilution of 1 in 20 (red line).

The secondary antibody used was FITC goat anti-rabbit at a dilution of 1 in 500. Rabbit monoclonal IgG was used as an isotype control (black line) and cells incubated in the absence of both primary and secondary antibody were used as a negative control (blue line).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD45 antibody [EP322Y] (ab40763)

Immunohistochemical staining of paraffin embedded human tonsil with purified ab40763 at a working dilution of 1/250.

The secondary antibody used is HRP goat anti-rabbit IgG H&L (ab97051) at 1/500. The sample is counterstained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0.

PBS was used instead of the primary antibody as the negative control (shown in the inset).
Immunocytochemistry/ Immunofluorescence - Anti-CD45 antibody [EP322Y] (ab40763)

Immunofluorescence staining of Jurkat cells (Human T cell leukemia cell line from peripheral blood) with purified ab40763 at a working dilution of 1/100, counterstained with DAPI.

The secondary antibody was Alexa Fluor^® 488 goat anti-rabbit (ab150077), used at a dilution of 1/1000. ab7291, a mouse anti-tubulin antibody (1/1000), was used to stain tubulin along with ab150120 (Alexa Fluor^® 594 goat anti-mouse, 1/1000), shown in the top right hand panel.

The cells were fixed in 4% PFA and permeabilized using 0.1% Triton X 100.

The negative controls are shown in bottom middle and right hand panels - for negative control 1, purified ab40763 was used at a dilution of 1/500 followed by an Alexa Fluor^® 594 goat anti-mouse antibody (ab150120) at a dilution of 1/500. For negative control 2, ab7291 (mouse anti-tubulin) was used at a dilution of 1/500 followed by an Alexa Fluor^® 488 goat anti-rabbit antibody (ab150077) at a dilution of 1/400.
Western blot - Anti-CD45 antibody [EP322Y] (ab40763)

Anti-CD45 antibody [EP322Y] (ab40763) at 1/5000 dilution (purified) + HuT-78 cell lysate at 20 µg

Secondary
HRP goat anti-rabbit IgG (H+L) at 1/50000 dilution

Predicted band size: 147 kDa
Observed band size: 200 kDa
why is the actual band size different from the predicted?

Blocking/Dilution buffer: 5% NFDM/TBST
Western blot - Anti-CD45 antibody [EP322Y] (ab40763)

Anti-CD45 antibody [EP322Y] (ab40763) at 1/5000 dilution (purified) + Raji (Human Burkitt's lymphoma cell line) cell lysate at 20 µg

Secondary
HRP goat anti-rabbit IgG (H+L) at 1/50000 dilution

Predicted band size: 147 kDa
Observed band size: 200 kDa why is the actual band size different from the predicted?

Blocking/Dilution buffer: 5% NFDM/TBST
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD45 antibody [EP322Y] (ab40763)

Unpurified ab40763 showing positive staining in normal spleen tissue.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD45 antibody [EP322Y] (ab40763)

Unpurified ab40763 showing positive staining in normal colon lymphoid cells tissue.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD45 antibody [EP322Y] (ab40763)

Unpurified ab40763 showing negative staining in skeletal muscle tissue.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD45 antibody [EP322Y] (ab40763)

Unpurified ab40763 showing negative staining in normal kidney tissue.
Western blot - Anti-CD45 antibody [EP322Y] (ab40763)

Anti-CD45 antibody [EP322Y] (ab40763) at 1/5000 dilution (unpurified, blocked with 3% milk) + Jurkat (Human T cell lymphoblast-like cell line) whole cell lysate at 10 µg

Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution

Developed using the ECL technique.

Performed under reducing conditions.

Predicted band size: 147 kDa
Observed band size: 200 kDa why is the actual band size different from the predicted?

Exposure time: 8 minutes

This blot was produced using a 3-8% Tris Acetate gel under the TA buffer system. The gel was run at 150V for 60 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes.

The membrane was then blocked for an hour using 3% milk before being incubated with ab40763 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution ab133406.
Other - Anti-CD45 antibody [EP322Y] (ab40763)

Equilibrium disassociation constant (KD)
Learn more about KD
Go here to learn more about KD:
https://www.abcam.com/index.html?pageconfig=resource&rid=15749

This product has been referenced in:

Quispe Calla NE et al. Exogenous oestrogen inhibits genital transmission of cell-associated HIV-1 in DMPA-treated humanized mice. J Int AIDS Soc 21:N/A (2018). Read more (PubMed: 29334191) »
Sherman H et al. A Novel Three-Dimensional Immune Oncology Model for High-Throughput Testing of Tumoricidal Activity. Front Immunol 9:857 (2018). Read more (PubMed: 29740450) »

See all 12 Publications for this product

ab40763 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。

レビューと Q&A

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レビューを送る質問を送る

1-10 of 10 Abreviews or Q&A

Abcam guarantees this product to work in the species/application used in this Abreview.

Application

Western blot

Sample

Human Cell lysate - whole cell (HL60 (human acute promyelocytic leukemia))

Gel Running Conditions

Reduced Denaturing (10% SDS-PAGE)

Loading amount

15 µg

Specification

HL60 (human acute promyelocytic leukemia)

Blocking step

Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

投稿 Oct 30 2018

Application

Immunocytochemistry/ Immunofluorescence

Sample

Mouse Cell (MEL (murine erythroleukemia cell))

Permeabilization

Yes - Tween-20

Specification

MEL (murine erythroleukemia cell)

Blocking step

BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: 25°C

Fixative

Acetone

Abcam user community

Verified customer

投稿 Oct 30 2018

Application

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

Sample

Human Tissue sections (Human breast cancer tissue (form MDA-MB-231 Xenogr)

Antigen retrieval step

Heat mediated - Buffer/Enzyme Used: pH 6.0 Citric Acid

Permeabilization

Yes - Tween-20

Specification

Human breast cancer tissue (form MDA-MB-231 Xenogr

Blocking step

BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: 25°C

Fixative

Paraformaldehyde

Abcam user community

Verified customer

投稿 Oct 29 2018

Application

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

Sample

Human Tissue sections (tonsil)

Antigen retrieval step

Heat mediated - Buffer/Enzyme Used: Tris/EDTA ph9

Permeabilization

Specification

tonsil

Fixative

Formaldehyde

Abcam user community

Verified customer

投稿 Sep 20 2018

Application

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

Sample

Human Tissue sections (Tonsil)

Antigen retrieval step

Heat mediated - Buffer/Enzyme Used: pH 9.0 EDTA

Permeabilization

Specification

Tonsil

Blocking step

PB ab64226 as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: RT°C

Fixative

Formaldehyde

Abcam user community

Verified customer

投稿 Mar 30 2018

Abcam guarantees this product to work in the species/application used in this Abreview.

Application

Immunocytochemistry/ Immunofluorescence

Sample

Human Cell (skin)

Permeabilization

Specification

skin

Blocking step

BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 24°C

Fixative

Formaldehyde

Abcam user community

Verified customer

投稿 Aug 26 2016

Application

Immunohistochemistry (Frozen sections)

Sample

Human Tissue sections (human small intestine tissue)

Specification

human small intestine tissue

Permeabilization

Fixative

Paraformaldehyde

Dr. Irit Shoval

Verified customer

投稿 Sep 19 2014

Application

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

Blocking step

BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: 21°C

Antigen retrieval step

Heat mediated - Buffer/Enzyme Used: Citric acid

Sample

Human Tissue sections (Tonsil)

Specification

Tonsil

Permeabilization

Fixative

Formaldehyde

Mr. Carl Hobbs

Verified customer

投稿 May 31 2014

Question

Customer kindly called to ask about the Abreview process and discuss difficulties they had been experiencing with ab10558.

Abcam community

Verified customer

Asked on Aug 09 2012

Answer

Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products. I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement of ab10558 with ab40763 the order number is xxx. To check the status of the order please contact our Customer Service team and reference this number. Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know. As there is a price discrepancy between these items I have also created credit note for this difference. Your credit note ID is xxxx. I have asked our accounting department to issue a credit note for you, which can be redeemed against the invoice of a future order by passing it on to your purchasing department. To avoid confusion, please ensure your accounts department is aware of how the credit note is being used. If you have questions on how to use the credit note, please contact our accounting department. Our accounting department can be contacted by email at us.credits@abcam.com or by telephone using the information at the Contact Us link in the top right corner of our website. Please refer to the credit note ID in any correspondence with our accounting department. The credit note ID is for your reference only and does not automatically guarantee the credit. 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.

Abcam Scientific Support

Answered on Aug 09 2012

Question

Would like to try ab40763 in ICC/IF.

Abcam community

Verified customer

Asked on Jul 23 2012

Answer

DISCOUNT CODE: ***
Expiration date: November 24, 2012

This code will give you 1 free primary antibodybefore the expiration date. To redeem this offer, please submit an Abreview for ab40763 in ICC and include this code in the “Additional Comments” section so we know the Abreview is for this promotion. Please remember that submission of the Abreview is sufficient for the discount code to become active. For more information on how to submit an Abreview, please visit the site: www.abcam.com/Abreviews.

The code will be active once the Abreview has been submitted and can be redeemed in one of the following ways: 1) Call to place your order and mention the code to our customer service department; 2) Include the code in your fax order; 3) Place your order on the web and enter the promotional code.

Any feedback that you can provide will be greatly appreciated, whether positive or negative. If you have any further questions, please do not hesitate to contact us. We look forward to receiving your Abreview and wish you luck with your research.

The terms and conditions applicable to this offer can be found here: www.abcam.com/collaborationdiscount.

Abcam Scientific Support

Answered on Jul 23 2012

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

カスタム・ソリューション & パートナーシップ

Anti-CD45 抗体 [EP322Y] (ab40763)

製品の概要

製品の特性

関連製品

アプリケーション

ターゲット情報

画像

データシートおよび資料

参考文献

This product has been referenced in:

レビューと Q&A

Western blot abreview for Anti-CD45 antibody [EP322Y]

Immunocytochemistry/ Immunofluorescence abreview for Anti-CD45 antibody [EP322Y]

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-CD45 antibody [EP322Y]

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-CD45 antibody [EP322Y]

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-CD45 antibody [EP322Y]

Immunocytochemistry/ Immunofluorescence abreview for Anti-CD45 antibody [EP322Y]

Immunohistochemistry (Frozen sections) abreview for Anti-CD45 antibody [EP322Y]

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-CD45 [EP322Y] antibody