Recombinant Anti-TAP2 antibody [EPR26237-82] KO Tested (ab307282)

Product name

Anti-TAP2 antibody [EPR26237-82]
See all TAP2 primary antibodies
Description

Rabbit monoclonal [EPR26237-82] to TAP2
Host species

Rabbit
Specificity

Not suitable for rat and mouse for WB and IHC-P.
Tested applications

Suitable for: IHC-P, ICC/IF, IP, WB, Flow Cyt (Intra)more details
Species reactivity

Reacts with: Human
Does not react with: Mouse, Rat
Immunogen

Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: Wild-type HeLa whole cell lysate. Jurkat, Raji, MOLT-4 and HT-1080 whole cell lysate. Untreated HeLa whole cell lysate. IFN gamma treated HeLa whole cell lysate. Human tonsil tissue lysate. IHC-P: Human tonsil, kidney, liver, breast and cerebrum tissue. Human breast cancer tissue. Wild -type HeLa cell pellet. ICC/IF: Wild-type HeLa cells. Flow Cyt (Intra): Wild-type HeLa cells. IP: IFN gamma treated HeLa whole cell lysate.
General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle.
Storage buffer

pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR26237-82
Isotype

IgG
Research areas

Alternative Versions
- Anti-TAP2 antibody [EPR26237-82] - BSA and Azide free (ab307283)
Compatible Secondaries
- Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (ab150081)
- Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
Related Products
- VeriBlot for IP Detection Reagent (HRP) (ab131366)
- Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed (ab150120)

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab307282 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-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.
ICC/IF		1/500.
IP		1/30.
WB		1/1000. Predicted molecular weight: 76 kDa.
Flow Cyt (Intra)		1/500.

Notes
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.
ICC/IF 1/500.
IP 1/30.
WB 1/1000. Predicted molecular weight: 76 kDa.
Flow Cyt (Intra) 1/500.

Function

Involved in the transport of antigens from the cytoplasm to the endoplasmic reticulum for association with MHC class I molecules. Also acts as a molecular scaffold for the final stage of MHC class I folding, namely the binding of peptide. Nascent MHC class I molecules associate with TAP via tapasin. Inhibited by the covalent attachment of herpes simplex virus ICP47 protein, which blocks the peptide-binding site of TAP. Inhibited by human cytomegalovirus US6 glycoprotein, which binds to the lumenal side of the TAP complex and inhibits peptide translocation by specifically blocking ATP-binding to TAP1 and prevents the conformational rearrangement of TAP induced by peptide binding. Inhibited by human adenovirus E3-19K glycoprotein, which binds the TAP complex and acts as a tapasin inhibitor, preventing MHC class I/TAP association.
Involvement in disease

Bare lymphocyte syndrome 1
Sequence similarities

Belongs to the ABC transporter superfamily. ABCB family. MHC peptide exporter (TC 3.A.1.209) subfamily.
Contains 1 ABC transmembrane type-1 domain.
Contains 1 ABC transporter domain.
Domain

The peptide-binding site is shared between the cytoplasmic loops of TAP1 and TAP2.
Cellular localization

Endoplasmic reticulum membrane. The transmembrane segments seem to form a pore in the membrane.
Target information above from: UniProt accession Q03519 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
Database links
- Entrez Gene: 6891 Human
- Omim: 170261 Human
- SwissProt: Q03519 Human
- Unigene: 502 Human
Alternative names
- ABC transporter, MHC 2 antibody
- ABC18 antibody
- ABCB3 antibody
- Antigen peptide transporter 2 antibody
- APT2 antibody
- ATP binding cassette, sub family B (MDR/TAP), member 3 antibody
- ATP-binding cassette sub-family B member 3 antibody
- D6S217E antibody
- Peptide supply factor 2 antibody
- Peptide transporter involved in antigen processing 2 antibody
- Peptide transporter PSF2 antibody
- Peptide transporter TAP2 antibody
- PSF 2 antibody
- PSF-2 antibody
- PSF2 antibody
- Really interesting new gene 11 protein antibody
- RING 11 antibody
- RING11 antibody
- TAP 2 antibody
- Tap2 antibody
- TAP2_HUMAN antibody
- Transporter 2 ATP binding cassette sub family B antibody
- Transporter 2, ABC (ATP binding cassette antibody
- Transporter 2, ATP binding cassette, sub family B (MDR/TAP) antibody
see all

Western blot - Anti-TAP2 antibody [EPR26237-82] (ab307282)

All lanes : Anti-TAP2 antibody [EPR26237-82] (ab307282) at 1/1000 dilution

Lane 1 : Wild-type HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate
Lane 2 : TAP2 knockout HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate
Lane 3 : Jurkat (human T cell leukemia T lymphocyte from peripheral blood) whole cell lysate
Lane 4 : Raji (human Burkitt's lymphoma B lymphocyte) whole cell lysate
Lane 5 : Untreated HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lane 6 : HeLa treated with 10 ng/ml IFN gamma for 16 hours whole cell lysate
Lane 7 : MOLT-4 (human lymphoblastic leukemia T lymphoblast) whole cell lysate
Lane 8 : HT-1080 (human fibrosarcoma epithelial cell) whole cell lysate
Lane 9 : Human tonsil tissue lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/100000 dilution

Predicted band size: 76 kDa
Observed band size: 75 kDa why is the actual band size different from the predicted?

Samples are non-boiled as boiling may cause protein aggregates.

The expression of TAP2 is upregulated in response to IFN gamma treatment (PMID: 1946428).

Blocking/dilution buffer: 5% NFDM/TBST.

Exposure time: 26 seconds.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TAP2 antibody [EPR26237-82] (ab307282)

Immunohistochemical analysis of paraffin-embedded human tonsil tissue labeling TAP2 with AB307282 at 1/2000 (0.257 ug/ml) followed by ready to use LeicaDS9800 (Bond® Polymer Refine Detection). Positive staining in human tonsil. The section was incubated with ab307282 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control: Secondary antibody is ready to use LeicaDS9800 (Bond® Polymer Refine Detection).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TAP2 antibody [EPR26237-82] (ab307282)

Immunohistochemical analysis of paraffin-embedded cell pellets labeling TAP2 with AB307282 at 1/2000 (0.257 ug/ml) followed by ready to use LeicaDS9800 (Bond® Polymer Refine Detection).

Positive staining in wild-type HeLa cell pellet and no staining in TAP2 knockout HeLa cell pellet.

The section was incubated with ab307282 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control: Secondary antibody is ready to use LeicaDS9800 (Bond® Polymer Refine Detection).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TAP2 antibody [EPR26237-82] (ab307282)

Immunohistochemical analysis of paraffin-embedded human breast cancer tissue labeling TAP2 with AB307282 at 1/2000 (0.257 ug/ml) followed by ready to use LeicaDS9800 (Bond® Polymer Refine Detection). Positive staining in human breast cancer (PMID: 22065046). The section was incubated with ab307282 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control: Secondary antibody is ready to use LeicaDS9800 (Bond® Polymer Refine Detection).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TAP2 antibody [EPR26237-82] (ab307282)

Immunohistochemical analysis of paraffin-embedded human cerebrum tissue labeling TAP2 with AB307282 at 1/2000 (0.257 ug/ml) followed by ready to use LeicaDS9800 (Bond® Polymer Refine Detection). Positive staining in human cerebrum. The section was incubated with ab307282 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control: Secondary antibody is ready to use LeicaDS9800 (Bond® Polymer Refine Detection).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TAP2 antibody [EPR26237-82] (ab307282)

Immunohistochemical analysis of paraffin-embedded human kidney tissue labeling TAP2 with AB307282 at 1/2000 (0.257 ug/ml) followed by ready to use LeicaDS9800 (Bond® Polymer Refine Detection). Positive staining in human kidney. The section was incubated with ab307282 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control: Secondary antibody is ready to use LeicaDS9800 (Bond® Polymer Refine Detection).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TAP2 antibody [EPR26237-82] (ab307282)

Immunohistochemical analysis of paraffin-embedded human liver tissue labeling TAP2 with AB307282 at 1/2000 (0.257 ug/ml) followed by ready to use LeicaDS9800 (Bond® Polymer Refine Detection). Positive staining in human liver. The section was incubated with ab307282 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control: Secondary antibody is ready to use LeicaDS9800 (Bond® Polymer Refine Detection).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TAP2 antibody [EPR26237-82] (ab307282)

Immunohistochemical analysis of paraffin-embedded human breast tissue labeling TAP2 with AB307282 at 1/2000 (0.257 ug/ml) followed by ready to use LeicaDS9800 (Bond® Polymer Refine Detection). Positive staining in human breast. The section was incubated with ab307282 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control: Secondary antibody is ready to use LeicaDS9800 (Bond® Polymer Refine Detection).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
Immunocytochemistry/ Immunofluorescence - Anti-TAP2 antibody [EPR26237-82] (ab307282)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized TAP2 KO HeLa (TAP2 knockout human cervical adenocarcinoma epithelial cell) ( ab265426) cells labeling TAP2 with AB307282 at 1/500 (1.028 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2 ug/ml) dilution (Green).

Confocal image showing endoplasmic reticulum staining in wildtype HeLa cells and no staining in TAP2 knockout HeLa cells.

Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8).

ab12223 Anti-KDEL mouse monoclonal antibody was used to counterstain tubulin at 1/200 dilution (5 ug/ml) dilution, followed by ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) at a 1/1000 dilution (2 ug/ml) (Red). The nuclear counterstain was DAPI (Blue).

Secondary antibody only control: Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 ug/ml) dilution.
Flow Cytometry (Intracellular) - Anti-TAP2 antibody [EPR26237-82] (ab307282)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Wild-type HeLa (human cervix adenocarcinoma epithelial cell, Right) / TAP2 knockout HeLa (Left) cells labeling TAP2 with AB307282 at 1/500 dilution (0.1ug) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/2000 dilution was used as the secondary antibody.

Positive staining on HeLa cells (ab255928), while no staining on TAP2 knockout HeLa cells (ab265426).
Immunoprecipitation - Anti-TAP2 antibody [EPR26237-82] (ab307282)

TAP2 was immunoprecipitated from 0.35 mg HeLa (human epithelial cell line from cervix adenocarcinoma) treated with 10 ng/ml IFN gamma for 16 hours, whole cell lysate 10 ug with AB307282 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using AB307282 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1: HeLa treated with 10 ng/ml IFN gamma for 16 hours, whole cell lysate 10 ug

Lane 2: AB307282 IP in HeLa treated with 10 ng/ml IFN gamma for 16 hours, whole cell lysate

Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab307282 in HeLa treated with 10ng/ml IFN gamma for 16 hours, whole cell lysate

Blocking and dilution buffer and concentration: 5% NFDM/TBST.

Exposure time: 1 seconds.

Input is non-boiled as boiling may cause protein aggregates.
Anti-TAP2 antibody [EPR26237-82] (ab307282)

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

Overview

Product name

Description

Host species

Specificity

Tested applications

Species reactivity

Immunogen

Positive control

General notes

Properties

Form

Storage instructions

Storage buffer

Purity

Clonality

Clone number

Isotype

Research areas

Associated products

Alternative Versions

Compatible Secondaries

Isotype control

Related Products

Applications

The Abpromise guarantee

Target

Function

Involvement in disease

Sequence similarities

Domain

Cellular localization

Database links

Alternative names

Images

Protocols

Datasheets and documents

SDS download

Datasheet download

Certificate of Compliance

References (0)

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