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Rabbit Recombinant Monoclonal CD27 antibody. Suitable for IHC-P, WB, ICC/IF, Flow Cyt (Intra) and reacts with Human samples. Cited in 18 publications.


Images

Flow Cytometry (Intracellular) - Anti-CD27 antibody [EPR8569] (AB131254), expandable thumbnail
  • Western blot - Anti-CD27 antibody [EPR8569] (AB131254), expandable thumbnail
  • Immunocytochemistry/ Immunofluorescence - Anti-CD27 antibody [EPR8569] (AB131254), expandable thumbnail
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD27 antibody [EPR8569] (AB131254), expandable thumbnail
  • Western blot - Anti-CD27 antibody [EPR8569] (AB131254), expandable thumbnail

Publications

Key facts

Isotype

IgG

Host species

Rabbit

Storage buffer

pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA

Form

Liquid

Clonality

Monoclonal

Immunogen

  • The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
IHC-PWBICC/IFFlow Cyt (Intra)
Human
Tested
Tested
Tested
Tested

Tested
Tested

Species

Human

Dilution info

1/1500

Notes

For unpurified use at 1/100 - 1/250.

Perform heat-mediated antigen retrieval before commencing with IHC staining protocol.

Tested
Tested

Species

Human

Dilution info

1/1000

Notes

For unpurified use at 1/1000 - 1/10000.

Tested
Tested

Species

Human

Dilution info

1/250 - 1/500

Notes

-

Tested
Tested

Species

Human

Dilution info

1/100 - 1/500

Notes

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

Associated Products

Select an associated product type

15 products for Alternative Product

1 products for Alternative Version

Target data

Function

Receptor for CD70/CD27L. May play a role in survival of activated T-cells. May play a role in apoptosis through association with SIVA1.

Alternative names

CD27 antigen, CD27L receptor, T-cell activation antigen CD27, T14, Tumor necrosis factor receptor superfamily member 7, CD27, TNFRSF7

Recommended products

Rabbit Recombinant Monoclonal CD27 antibody. Suitable for IHC-P, WB, ICC/IF, Flow Cyt (Intra) and reacts with Human samples. Cited in 18 publications.

Alternative names

CD27 antigen, CD27L receptor, T-cell activation antigen CD27, T14, Tumor necrosis factor receptor superfamily member 7, CD27, TNFRSF7

Key facts

Isotype

IgG

Form

Liquid

Clonality

Monoclonal

Immunogen
  • The exact immunogen used to generate this antibody is proprietary information.
Clone number

EPR8569

Purification technique

Affinity purification Protein A

Dissociation constant

7.9 x 10-11 M

Concentration
Loading...

Storage

Shipped at conditions

Blue Ice

Appropriate short-term storage duration

1-2 weeks

Appropriate short-term storage conditions

+4°C

Appropriate long-term storage conditions

-20°C

Aliquoting information

Upon delivery aliquot

Storage information

Stable for 12 months at -20°C

Notes

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.

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 batch production

For more information, read more on recombinant antibodies.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity summary

CD27 also known as Tumor Necrosis Factor Receptor Superfamily Member 7 (TNFRSF7) is a protein chiefly involved in T cell activation and proliferation. It belongs to the TNF-receptor superfamily and weighs approximately 50 kDa. CD27 is expressed mainly on T lymphocytes and to a lesser extent on B cells and natural killer cells. Researchers often study CD27 and its ligand CD70 to understand immune responses. Tools such as anti-CD27 percp 7h21 monoclonal antibodies and APC-conjugated forms help detect this protein in various assays.

Biological function summary

One finds CD27 at the heart of cellular immune responses. It does not function alone; instead it typically forms part of a receptor-ligand complex with CD70 which influences T cell and B cell activities. This interaction promotes cell survival and plays a role in the activation of NF-kB and MAPK8/JNK signaling pathways which are essential for immune response regulation. These pathways ensure that the body's immune cells can efficiently respond to pathogens.

Pathways

CD27 actively integrates into the NF-kB and mTOR signaling pathways which are important for regulating immune and inflammatory responses. Through these pathways CD27 works closely with proteins like TRAF2 and TRAF3 which are adapter molecules that transmit signals from the receptor. These signaling pathways provide a framework for understanding how CD27 and associated proteins work together to control immune cell behavior facilitating coordinated immune defense mechanisms.

Associated diseases and disorders

CD27 has significant implications in autoimmune diseases and cancer. For example aberrant activation of CD27 may contribute to conditions like lymphoma and autoimmune diseases such as lupus. The CD70-CD27 signaling axis when dysregulated can lead to uncontrolled cell proliferation or apoptosis linking it to these pathologies. Understanding the interplay between CD27 and proteins like Bcl-2 which influences apoptosis helps pave the way for therapeutic strategies targeting these diseases.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

13 product images

  • Flow Cytometry (Intracellular) - Anti-CD27 antibody [EPR8569] (ab131254), expandable thumbnail

    Flow Cytometry (Intracellular) - Anti-CD27 antibody [EPR8569] (ab131254)

    Flow Cytometry analysis of Ramos cells labelling CD27 with purified ab131254 at 1/300 (red). Cells were fixed with 2% paraformaldehyde. A FITC-conjugated goat anti-rabbit IgG (1/150) was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG. Blue - Unlabelled control, cells without incubation with primary and secondary antibodies.

  • Western blot - Anti-CD27 antibody [EPR8569] (ab131254), expandable thumbnail

    Western blot - Anti-CD27 antibody [EPR8569] (ab131254)

    Lanes 1 - 4: Merged signal (red and green). Green - ab131254 observed at 35 kDa. Red - loading control Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55kDa.

    ab131254 was shown to react with CD27 in wild-type Raji cells in western blot with loss of signal observed in CD27 knockout sample. Wild-type and CD27 knockout Raji cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with ab131254 and Anti-alpha Tubulin antibody [DM1A] - Loading Control 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 (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

    All lanes: Western blot - Anti-CD27 antibody [EPR8569] (ab131254) at 1/1000 dilution

    Lane 1: Wild-type Raji (Human Burkitt's lymphoma cell line) whole cell lysate at 20 µg

    Lane 2: CD27 knockout Raji (Human Burkitt's lymphoma cell line) whole cell lysate at 20 µg

    Lane 3: Ramos (Human Burkitt's lymphoma cell line) whole cell lysate at 20 µg

    Lane 4: Human brain tissue lysate at 20 µg

    Performed under reducing conditions.

    Predicted band size: 29 kDa

    Observed band size: 35 kDa

  • Immunocytochemistry/ Immunofluorescence - Anti-CD27 antibody [EPR8569] (ab131254), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-CD27 antibody [EPR8569] (ab131254)

    ab131254 staining CD27 in Raji cells, with negative expression in A375 cells. The cells were fixed with 100% methanol (5 min), permeabilised with 0.1% Triton x-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab131254 at 1 μg/ml and Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291, Mouse monoclonal [DM1A] to alpha Tubulin at 0.5 μg/ml. Cells were then incubated with Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 488), pre-adsorbed at 1/1000 dilution (shown in green) and Goat Anti-Mouse IgG H&L (Alexa Fluor® 647) preadsorbed ab150119, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor® 647), pre-adsorbed at 1/1000 dilution (shown in red). Nuclear DNA was labelled with DAPI (shown in blue).

    Image was acquired with a confocal microscope (Leica-Microsystems TCS SP8) and a single confocal section is shown.

    This product also work with 4% formaldehyde (10 min) fixation under the same testing conditions.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD27 antibody [EPR8569] (ab131254), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD27 antibody [EPR8569] (ab131254)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human tonsil tissue labelling CD27 with purified ab131254 at 1/1500. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. Goat Anti-Rabbit IgG H&L (HRP) ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500).

    Negative control using PBS instead of primary antibody.

    Counterstained with hematoxylin.

  • Western blot - Anti-CD27 antibody [EPR8569] (ab131254), expandable thumbnail

    Western blot - Anti-CD27 antibody [EPR8569] (ab131254)

    Blocking and dilution buffer: 5% NFDM/TBST.

    All lanes: Western blot - Anti-CD27 antibody [EPR8569] (ab131254) at 1/5000 dilution

    All lanes: NAMALWA (human Burkitt's lymphoma cell line) cell lysate at 20 µg

    Secondary

    All lanes: Peroxidase-conjugated goat anti-rabbit IgG, (H+L) at 1/1000 dilution

    Predicted band size: 137 kDa, 29 kDa, 31 kDa, 35 kDa, 42 kDa, 52 kDa, 55 kDa, 57 kDa

    Observed band size: 185 kDa, 55 kDa

  • Western blot - Anti-CD27 antibody [EPR8569] (ab131254), expandable thumbnail

    Western blot - Anti-CD27 antibody [EPR8569] (ab131254)

    Blocking and dilution buffer: 5% NFDM/TBST.

    All lanes: Western blot - Anti-CD27 antibody [EPR8569] (ab131254) at 1/1000 dilution

    Lane 1: Human lymph node tissue lysate at 20 µg

    Lane 2: Human fetal spleen tissue lysate at 20 µg

    Secondary

    All lanes: Peroxidase-conjugated goat anti-rabbit IgG, (H+L) at 1/1000 dilution

    Predicted band size: 29 kDa

    Observed band size: 55 kDa

  • Immunocytochemistry/ Immunofluorescence - Anti-CD27 antibody [EPR8569] (ab131254), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-CD27 antibody [EPR8569] (ab131254)

    Immunocytochemistry/Immunofluorescence analysis of Jurkat cells labelling CD27 with purified ab131254 at 1/500. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain. Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291, a mouse anti-tubulin (1/1000) and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/500) were also used.

    Control 1: primary antibody (1/500) and secondary antibody, Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/500).

    Control 2: Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (1/1000) and secondary antibody, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/500).

  • Western blot - Anti-CD27 antibody [EPR8569] (ab131254), expandable thumbnail

    Western blot - Anti-CD27 antibody [EPR8569] (ab131254)

    All lanes: Western blot - Anti-CD27 antibody [EPR8569] (ab131254) at 1/1000 dilution

    Lane 1: Ramos (human Burkitt's lymphoma cell line) cell lysate at 10 µg

    Lane 2: Human lymph node cell lysate at 10 µg

    Secondary

    All lanes: HRP labelled Goat anti Rabbit IgG at 1/2000 dilution

    Predicted band size: 29 kDa

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD27 antibody [EPR8569] (ab131254), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD27 antibody [EPR8569] (ab131254)

    Immunohistochemistry (Formalin/PFA-fixed paraffin embedded sections) analysis of human stomach tissue labelling CD27 with ab131254 at a dilution of 1/100.

    Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD27 antibody [EPR8569] (ab131254), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD27 antibody [EPR8569] (ab131254)

    Immunohistochemistry (Formalin/PFA-fixed paraffin embedded sections) analysis of human tonsil tissue labelling CD27 with ab131254 at a dilution of 1/100.

    Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

  • OI-RD Scanning - Anti-CD27 antibody [EPR8569] (ab131254), expandable thumbnail

    OI-RD Scanning - Anti-CD27 antibody [EPR8569] (ab131254)

    We have systematically measured KD (the equilibrium dissociation constant between the antibody and its antigen), of more than 840 recombinant antibodies to assess not only their individual KD values but also to see the average affinity of antibody.
    Based on the comparison with published literature values for mouse monoclonal antibodies, Recombinant antibodies appear to be on average 1-2 order of magnitude higher affinity.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD27 antibody [EPR8569] (ab131254), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD27 antibody [EPR8569] (ab131254)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human skeletal muscle showing negative staining with purified ab131254 at 1/1800. Heat mediated antigen retrieval was performed using Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) ab93684 (Tris/EDTA buffer, pH 9.0). Goat Anti-Rabbit IgG H&L (HRP polymer) ab214880, a Goat Anti-Rabbit IgG H&L (HRP polymer) was used as the secondary antibody (1/500).

    Counterstained with hematoxylin.

    Negative control: No staining on human skeletal muscle. The section was incubated with ab131254 at 4°C overnight.

  • Flow Cytometry (Intracellular) - Anti-CD27 antibody [EPR8569] (ab131254), expandable thumbnail

    Flow Cytometry (Intracellular) - Anti-CD27 antibody [EPR8569] (ab131254)

    Flow cytometry staining of human peripheral blood mononuclear cells (PBMCs) with ab131254 (right) or Recombinant Rabbit IgG, monoclonal [EPR25A] - Isotype Control (left). Cells were fixed and permeabilised with BD Cytofix/Cytoperm™ for 20 min. PBMCs were incubated for 30 min at 22°C in 1x PBS containing 10 µg/ml human IgG and 10 % normal goat serum to block FC receptors and non-specific protein-protein interaction followed by the antibody ab131254 or Recombinant Rabbit IgG, monoclonal [EPR25A] - Isotype Control (1x 106 in 100 µl at 0.04 μg/ml (1/52750)) for 30 min at 4°C . The cells were simultaneously stained with CD4.

    The secondary antibody Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed was incubated at 1/4000 for 30min at 4°C

    Acquisition of >30000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter. Events were gated on viable cells.

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Product protocols

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