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
  • Host species

    Mouse
  • 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

    • Antigen Presenting Molecule antibody
    • Cw*1701 antibody
    • D6S204 antibody
    • FLJ27082 antibody
    • HLA A antibody
    • HLA B antibody
    • HLA C antibody
    • HLA class 1 A antibody
    • HLA class 1 B antibody
    • HLA class 1 C antibody
    • HLA JY3 antibody
    • HLAA antibody
    • HLAB antibody
    • HLAC antibody
    • HLC C antibody
    • Human Leucocyte Antigen C antibody
    • Major Histocompatibility Complex Class I A antibody
    • Major Histocompatibility Complex Class I B antibody
    • Major Histocompatibility Complex Class I C antibody
    • Major histocompatibility complex, class I, A + B + C antibody
    • MHC class I antigen A*1 antibody
    • MHC Class I Antigen antibody
    • MHC class I antigen B*7 antibody
    • MHC class I antigen Cw*1 antibody
    • MHC class I HLA A antibody
    • MHC class I HLA B antibody
    • MHC class I HLA C antibody
    • MHC HLA ABC antibody
    • PSORS1 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:

  • Boxberg M  et al. Composition and Clinical Impact of the Immunologic Tumor Microenvironment in Oral Squamous Cell Carcinoma. J Immunol 202:278-291 (2019). Read more (PubMed: 30530592) »
  • Watanabe S  et al. Mutational activation of the epidermal growth factor receptor down-regulates major histocompatibility complex class I expression via the extracellular signal-regulated kinase in non-small cell lung cancer. Cancer Sci 110:52-60 (2019). Read more (PubMed: 30390416) »
See all 66 Publications for this product

Customer reviews and Q&As

1-10 of 12 Abreviews or Q&A

Application
Immunohistochemistry (Frozen sections)
Sample
Human Tissue sections (Melanoma metastasis)
Permeabilization
No
Specification
Melanoma metastasis
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: 21°C
Fixative
Acetone

Abcam user community

Verified customer

Submitted Mar 06 2019

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (Tonsil)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citrate pH 6
Permeabilization
No
Specification
Tonsil
Blocking step
Dako Protein Block X0909 as blocking agent for 15 minute(s) · Concentration: .25% · Temperature: 25°C
Fixative
Formaldehyde

Abcam user community

Verified customer

Submitted Dec 20 2018

Application
Immunohistochemistry (Frozen sections)
Sample
Human Tissue sections (LCL cells)
Permeabilization
No
Specification
LCL cells
Blocking step
PB ab64226 as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: RT°C
Fixative
Neutral buffered formalin

Abcam user community

Verified customer

Submitted Feb 06 2018

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

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

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

Abcam user community

Verified customer

Submitted Jan 06 2014

Answer


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

Read More
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

Abcam user community

Verified customer

Submitted Oct 04 2012

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

Abcam user community

Verified customer

Submitted Jul 18 2012

Answer

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 antibodies are created against. However, if you are able to find the proteins sequences I would gladly do an alignment. Should you wish to perform the alignment for yourself, when homology is less than 85% we usually believe that there is minimal cross reactivity.

I hope this information is helpful. Please let me know if you have any questions.

Read More

1-10 of 12 Abreviews or Q&A

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

Sign up