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    cd80-antibody-epr11572-ab134120.pdf

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Immunology Adaptive Immunity B Cells CD
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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-CD80 antibody [EPR1157(2)] (ab134120)

  • Datasheet
  • SDS
Reviews (2) Submit a question References (15)

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Flow Cytometry - Anti-CD80 antibody [EPR1157(2)] (ab134120)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD80 antibody [EPR1157(2)] (ab134120)
  • Western blot - Anti-CD80 antibody [EPR1157(2)] (ab134120)
  • Western blot - Anti-CD80 antibody [EPR1157(2)] (ab134120)
  • Western blot - Anti-CD80 antibody [EPR1157(2)] (ab134120)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD80 antibody [EPR1157(2)] (ab134120)
  • Anti-CD80 antibody [EPR1157(2)] (ab134120)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR1157(2)] to CD80
  • Suitable for: Flow Cyt, WB, IHC-P
  • Knockout validated
  • Reacts with: Human

Conjugates logo Related conjugates and formulations

Carrier Free

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Knockout
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Human CD80 knockout Raji cell line (ab273873)

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Overview

  • Product name

    Anti-CD80 antibody [EPR1157(2)]
    See all CD80 primary antibodies
  • Description

    Rabbit monoclonal [EPR1157(2)] to CD80
  • Host species

    Rabbit
  • Tested applications

    Suitable for: Flow Cyt, WB, IHC-Pmore details
    Unsuitable for: ICC/IF
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: Raji and Ramos cell lysates. IHC-P: human tonsil Tissue Flow Cyt: Raji cells
  • 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.

    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.

    Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

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. Stable for 12 months at -20°C.
  • Storage buffer

    pH: 7.2
    Preservative: 0.01% Sodium azide
    Constituents: 40% Glycerol (glycerin, glycerine), 0.05% BSA, 59% PBS
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR1157(2)
  • Isotype

    IgG
  • Research areas

    • Immunology
    • Adaptive Immunity
    • B Cells
    • CD
    • Stem Cells
    • Hematopoietic Progenitors
    • Lymphoid
    • B Lymphocytic Lineage
    • Stem Cells
    • Hematopoietic Progenitors
    • Myeloid
    • Dendritic Cell Lineage
    • Stem Cells
    • Hematopoietic Progenitors
    • Myeloid
    • Monocytic Lineage

Associated products

  • Alternative Versions

    • Anti-CD80 antibody [EPR1157(2)] - Low endotoxin, Azide free (ab224797)
    • Anti-CD80 antibody [EPR1157(2)] - BSA and Azide free (ab271905)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (ab150081)
  • Conjugation kits

    • PE / R-Phycoerythrin Conjugation Kit - Lightning-Link® (ab102918)
    • APC Conjugation Kit - Lightning-Link® (ab201807)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • KO cell lines

    • Human CD80 knockout Raji cell line (ab273873)
  • KO cell lysates

    • Human CD80 knockout Raji cell lysate (ab273827)
  • Positive Controls

    • Raji membrane extract lysate (ab30126)
    • Ramos whole cell lysate (ab3955)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab134120 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 at an assay dependent concentration.
WB
1/1000 - 1/10000. Detects a band of approximately 60 kDa (predicted molecular weight: 33 kDa).

For unpurified use at 1/500.

IHC-P (2)
1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Notes
Flow Cyt
Use at an assay dependent concentration.
WB
1/1000 - 1/10000. Detects a band of approximately 60 kDa (predicted molecular weight: 33 kDa).

For unpurified use at 1/500.

IHC-P
1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Application notes
Is unsuitable for ICC/IF.

Target

  • Function

    Involved in the costimulatory signal essential for T-lymphocyte activation. T-cell proliferation and cytokine production is induced by the binding of CD28 or CTLA-4 to this receptor.
  • Tissue specificity

    Expressed on activated B-cells, macrophages and dendritic cells.
  • Sequence similarities

    Contains 1 Ig-like C2-type (immunoglobulin-like) domain.
    Contains 1 Ig-like V-type (immunoglobulin-like) domain.
  • Cellular localization

    Membrane.
  • Target information above from: UniProt accession P33681 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 941 Human
    • Omim: 112203 Human
    • SwissProt: P33681 Human
    • Unigene: 838 Human
    • Alternative names

      • Activation B7-1 antigen antibody
      • B lymphocyte activation antigen B7 antibody
      • B7 antibody
      • B7-1 antibody
      • B7-1 antigen antibody
      • B7.1 antibody
      • BB1 antibody
      • CD28 antigen ligand 1 antibody
      • CD28LG antibody
      • CD28LG1 antibody
      • CD80 antibody
      • CD80 antigen (CD28 antigen ligand 1, B7-1 antigen) antibody
      • CD80 antigen antibody
      • CD80 molecule antibody
      • CD80_HUMAN antibody
      • Costimulatory factor CD80 antibody
      • costimulatory molecule variant IgV-CD80 antibody
      • CTLA-4 counter-receptor B7.1 antibody
      • LAB7 antibody
      • T-lymphocyte activation antigen CD80 antibody
      see all

    Images

    • Flow Cytometry - Anti-CD80 antibody [EPR1157(2)] (ab134120)
      Flow Cytometry - Anti-CD80 antibody [EPR1157(2)] (ab134120)
      Flow Cytometry analysis of Raji (human Burkitt's lymphoma B lymphocyte) cells labeling CD80 with purified ab134120 at 1:190 dilution (10.33 µg/ml) - Red. Cells were fixed with 4% paraformaldehyde . A Goat anti rabbit IgG (Alexa Fluor® 488, ab150081) secondary antibody was used at 1:2000 dilution. Isotype control - Rabbit monoclonal IgG (ab172730) - Black. Unlabeled control - Blue. Untreated cells - Green
    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD80 antibody [EPR1157(2)] (ab134120)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD80 antibody [EPR1157(2)] (ab134120)

      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human tonsil tissue labelling CD80 with ab134120 at a dilution of 1/1000. Heat mediated antigen retrieval was performed using Trs/EDTA buffer, pH9 (ab93684). ImmunoHistoProbe one step HRP Polymer was used. Counter stained with hematoxylin.

      The image shows cytoplasmic and membrane staining.

    • Western blot - Anti-CD80 antibody [EPR1157(2)] (ab134120)
      Western blot - Anti-CD80 antibody [EPR1157(2)] (ab134120)
      All lanes : Anti-CD80 antibody [EPR1157(2)] (ab134120) at 1/1000 dilution

      Lane 1 : Wild-type Raji cell lysate
      Lane 2 : CD80 knockout Raji cell lysate
      Lane 3 : Ramos cell lysate

      Lysates/proteins at 20 µg per lane.

      Performed under reducing conditions.

      Predicted band size: 33 kDa
      Observed band size: 55 kDa why is the actual band size different from the predicted?



      Lanes 1 - 3: Merged signal (red and green). Green - ab134120 observed at 55 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55kDa.

      ab134120 was shown to react with CD80 in wild-type Raji cells in western blot with loss of signal observed in CD80 knockout sample. Wild-type and CD80 knockout Raji cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with ab134120 and ab7291 (Mouse anti-Alpha Tubulin [DM1A]) overnight at 4°C at a 1 in 1000 Dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

    • Western blot - Anti-CD80 antibody [EPR1157(2)] (ab134120)
      Western blot - Anti-CD80 antibody [EPR1157(2)] (ab134120)
      Anti-CD80 antibody [EPR1157(2)] (ab134120) at 1/2500 dilution (purified) + Raji cell lysate at 20 µg

      Secondary
      Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

      Predicted band size: 33 kDa
      Observed band size: 60 kDa why is the actual band size different from the predicted?



      Blocking buffer and concentration: 5% NFDM/TBST.

      Diluting buffer and concentration: 5% NFDM/TBST.

    • Western blot - Anti-CD80 antibody [EPR1157(2)] (ab134120)
      Western blot - Anti-CD80 antibody [EPR1157(2)] (ab134120)
      All lanes : Anti-CD80 antibody [EPR1157(2)] (ab134120) at 1/1000 dilution (unpurified)

      Lane 1 : Raji cell lysate
      Lane 2 : Ramos cell lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : Goat anti-rabbit HRP conjugated antibody at 1/2000 dilution

      Predicted band size: 33 kDa
      Observed band size: 60 kDa why is the actual band size different from the predicted?

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD80 antibody [EPR1157(2)] (ab134120)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD80 antibody [EPR1157(2)] (ab134120)

      Tissue Microarrays stained for "Anti-CD80 antibody [EPR1157(2)]” using "ab134120" in immunohistochemical analysis. This table provides a detailed overview of positive (tick mark) and negative (cross mark) staining per sample type tested. The sections were pre-treated using Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0) for 20 minutes. The sections were incubated with ab134120 for 30 mins at room temperature followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

    • Anti-CD80 antibody [EPR1157(2)] (ab134120)
      Anti-CD80 antibody [EPR1157(2)] (ab134120)

    Protocols

    • Western blot protocols
    • Immunohistochemistry protocols

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    References (15)

    Publishing research using ab134120? Please let us know so that we can cite the reference in this datasheet.

    ab134120 has been referenced in 15 publications.

    • Zhang L  et al. Single-Cell Analyses Inform Mechanisms of Myeloid-Targeted Therapies in Colon Cancer. Cell 181:442-459.e29 (2020). PubMed: 32302573
    • Kuwabara S  et al. Prognostic relevance of tertiary lymphoid organs following neoadjuvant chemoradiotherapy in pancreatic ductal adenocarcinoma. Cancer Sci 110:1853-1862 (2019). PubMed: 30997706
    • Cui CY  et al. Skewed macrophage polarization in aging skeletal muscle. Aging Cell 18:e13032 (2019). PubMed: 31478346
    • Nascimento Da Conceicao V  et al. Loss of Ca2+ entry via Orai-TRPC1 induces ER stress, initiating immune activation in macrophages. J Cell Sci 133:N/A (2019). PubMed: 31722977
    • Jaiswal SR  et al. CTLA4Ig-based reduced intensity conditioning and donor lymphocyte infusions for haploidentical transplantation in refractory aggressive B-cell lymphoma relapsing after an autograft: Early results from a pilot study. Exp Hematol 77:26-35.e1 (2019). PubMed: 31449833
    View all Publications for this product

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review

    Filter by Application

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    1-2 of 2 Abreviews

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-CD80 antibody [EPR1157(2)]

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Human Tissue sections (tonsil)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Tris/EDTA pH 9.0
    Permeabilization
    No
    Specification
    tonsil
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 15% · Temperature: RT°C
    Fixative
    10% NBF
    Read More
    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

    Submitted May 16 2019

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-CD80 antibody [EPR1157(2)]

    Inconclusive
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Mouse Tissue sections (tumor)
    Antigen retrieval step
    Heat mediated
    Permeabilization
    No
    Specification
    tumor
    Fixative
    Formaldehyde
    Read More
    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.

    MISS. Sylvie Colin

    Verified customer

    Submitted Oct 20 2017

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