Anti-HLA Class 1 ABC antibody [EMR8-5] (ab70328)

Overview

  • Product name
    Anti-HLA Class 1 ABC antibody [EMR8-5]
    See all HLA Class 1 ABC primary antibodies
  • Description
    Mouse monoclonal [EMR8-5] to HLA Class 1 ABC
  • Specificity
    ab70328 reacts with the heavy chains of human HLA class 1 A, -B, and -C.
  • Tested applications
    Suitable for: Flow Cyt, ICC/IF, WB, IHC-P, IHC-Frmore details
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Recombinant HLA-A*2402 extracellular domain

Properties

  • Form
    Liquid
  • Storage instructions
    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term.
  • Storage buffer
    Preservative: 0.05% Sodium Azide
    Constituents: PBS, pH 7.4
  • Concentration information loading...
  • Purity
    Protein A purified
  • Clonality
    Monoclonal
  • Clone number
    EMR8-5
  • Myeloma
    NS1
  • Isotype
    IgG1
  • Light chain type
    kappa
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab70328 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
Flow Cyt Use 1-2µg for 106 cells. (methanol or paraformaldehyde fixed cells)



ab170190-Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
ICC/IF 1/100.
WB Use at an assay dependent concentration.
IHC-P 1/50 - 1/100.
IHC-Fr Use at an assay dependent concentration. PubMed: 23766645

Target

  • Relevance
    HLA-ABC antigen is present on most nucleated cells of humans and Old World monkeys. HLA CLass I is involved in the presentation of foreign antigens to the immune system.
  • Cellular localization
    Cell Membrane; Single-pass type I membrane protein
  • Database links
  • Alternative names
    • HLA A antibody
    • HLA B antibody
    • HLA C antibody
    • HLA class 1 A antibody
    • HLA class 1 B antibody
    • HLA class 1 C antibody
    • Major histocompatibility complex, class I, A + B + C antibody
    • MHC class I HLA A antibody
    • MHC class I HLA B antibody
    • MHC class I HLA C antibody
    • MHC HLA ABC antibody
    see all

Images

  • Immunohistochemical analysis of murine lung with Human MDA-MB231 breast cancer cells, staining HLA Class 1 ABC with ab70328.

    Tissue was fixed with formaldehyde and blocked with blocking agent for 4 minutes at 36°C; antigen retrieval was by heat mediation. Samples were incubated with primary antibody (1/100 in diluent) for 24 minutes at 36°C. An undiluted HRP-conjugated goat anti-mouse polyclonal IgG was used as the secondary antibody.

    See Abreview

  • All lanes : Anti-HLA Class 1 ABC antibody [EMR8-5] (ab70328)

    Lane 1 : A*2402
    Lane 2 : A*0101
    Lane 3 : A*1101
    Lane 4 : A*0201
    Lane 5 : A*0207
    Lane 6 : A2402
    Lane 7 : B*0702
    Lane 8 : B*0801
    Lane 9 : B*1501
    Lane 10 : B*3501
    Lane 11 : B*4001
    Lane 12 : B*4002
    Lane 13 : B*4006
    Lane 14 : B*4403
    Lane 15 : Cw*0102
    Lane 16 : Cw*0801
    Lane 17 : Cw*1202
    Lane 18 : Cw*1502

    Western blot analysis of ab70328 reactivity using recominant HLA heavy chains.

    Lanes 1-6: HLA-A

    Lanes 7-14: HLA-B

    Lanes 15-18: HLA-C

  • All lanes : Anti-HLA Class 1 ABC antibody [EMR8-5] (ab70328)

    Lane 1 : Human oral cancer cell line OSC20
    Lane 2 : HLA-A2402 gene transfected OSC20
    Lane 3 : Human Leukemia cell line K562
    Lane 4 : Recombinant HLA-A2402 heavy chain
  • ab70328 staining HLA Class 1 ABC in Human Glioblastoma tumour grown in mouse brain by ICC/IF (Immunocytochemistry/immunofluorescence).

    Cells were fixed with formaldehyde, permeabilized with Triton X-100 and blocked with 10% serum for 30 minutes at room temperature. Samples were incubated with primary antibody (1/100 in diluent) for 16 hours at 4°C. An AlexaFluor®568-conjugated donkey monoclonal (1/400) was used as the secondary antibody.

    See Abreview

  • Overlay histogram showing Jurkat cells stained with ab70328 (red line). The cells were fixed with methanol (5 min) and incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab70328, 1µg/1x106 cells) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in Jurkat cells fixed with 4% paraformaldehyde (10 min) used under the same conditions.

    Please note that Abcam do not have data for use of this antibody on non-fixed cells. We welcome any customer feedback.

References

This product has been referenced in:
  • Mordoh J  et al. Phase II Study of Adjuvant Immunotherapy with the CSF-470 Vaccine Plus Bacillus Calmette-Guerin Plus Recombinant Human Granulocyte Macrophage-Colony Stimulating Factor vs Medium-Dose Interferon Alpha 2B in Stages IIB, IIC, and III Cutaneous Melanoma Patients: A Single Institution, Randomized Study. Front Immunol 8:625 (2017). IHC-P ; Human . Read more (PubMed: 28620382) »
  • Pavesi A  et al. A 3D microfluidic model for preclinical evaluation of TCR-engineered T cells against solid tumors. JCI Insight 2:N/A (2017). IHC-P ; Human . Read more (PubMed: 28614795) »

See all 34 Publications for this product

Customer reviews and Q&As

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (Colon/Tonsil)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: pH6 20 mins 100 oC
Permeabilization
No
Specification
Colon/Tonsil
Blocking step
No blocking step used for 10 minute(s) · Concentration: 3% · Temperature: 37°C
Fixative
Formaldehyde
Username

Mr. Christopher Bagnall

Verified customer

Submitted May 30 2016

Application
Western blot
Sample
Human Cell lysate - whole cell (PBMC)
Gel Running Conditions
Non-reduced Denaturing
Loading amount
10 µg
Specification
PBMC
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 4% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Nov 01 2015

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: citrate, pH6, 100C for 20 min
Sample
Human Tissue sections (tonsil, colon, colon cancer)
Specification
tonsil, colon, colon cancer
Permeabilization
No
Fixative
Formaldehyde
Username

Abcam user community

Verified customer

Submitted Jan 06 2014


This clone has been shown to have a kappa light chain.

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (lung with human MDA-MB231 breast cancer cells)
Antigen retrieval step
Heat mediated
Permeabilization
No
Specification
lung with human MDA-MB231 breast cancer cells
Blocking step
U ultraview DAB as blocking agent for 4 minute(s) · Concentration: 0% · Temperature: 36°C
Fixative
Formaldehyde
Username

Abcam user community

Verified customer

Submitted Oct 04 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (Human Glioblastoma tumour grown in mouse brain)
Specification
Human Glioblastoma tumour grown in mouse brain
Fixative
Formaldehyde
Permeabilization
Yes - Triton X-100
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: RT°C
Username

Abcam user community

Verified customer

Submitted Jul 18 2012



Es freut mich zu hören, dass Sie unser Angebot unseren Antikörper ab70328an Xenografts zu testen, annehmen möchten. Wie besprochen, können Sie den Gutscheincode zum Kauf eines weiteren Antikörpers einsetzen, sobald Si...

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Vielen Dank für Ihre Anfrage.

Bei über80000 Produkten, die wir derzeit im Katalog haben, können wir aus logistischen Gründen leider keine kostenlosen Proben zu Testzwecken anbieten. Hier bei Abcam verfahren wir so, dass wi...

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Thank you for contacting Abcam.

The products ab3283 and ab70328 have not been tested for cross reactivity withcynomolgus monkeys. I was unable to find the associated cynomolgis proteins to the human proteins that many of the CD47 and HLAC1 a...

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Thank you for contacting us. For ab53134, the antigen retrieval protocol is as follows: - Bring 500 - 2000 ml 10 mM citrate buffer (pH6.0) to the boil in a stainless steel pressure cooker. - Put the slide into staining rack and lower into pressure cook...

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

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