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AB281360

Human CD27A knockout Raji cell lysate

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CD27 KO cell lysate available now. KO validated by Next Generation Sequencing, Western blot. Free of charge wild type control included. Knockout achieved by CRISPR/Cas9 X = 10 bp deletion Frameshift: 100%.
3 Images
Western blot - Human CD27A knockout Raji cell lysate (AB281360)
  • WB

Lab

Western blot - Human CD27A knockout Raji cell lysate (AB281360)

Lane 1 : Wild-type Raji cell lysate (20 ug)
Lane 2 : CD27A knockout Raji cell lysate (20 ug)
Lane 3 : Ramos cell lysate (20 ug)
Lane 4 : Human brain cell lysate (20 ug)

ab133761 was shown to react with CD27 in wild-type Raji cells in western blot. Loss of signal was observed when CD27 knockout cell line ab274910 (knockout cell lysate ab281360) was used. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with ab133761 overnight at 4 °C at a 1 in 1000 dilution. Blots were incubated with HRP conjugated Goat anti-Rabbit (H+L) secondary antibody at 1 in 5000 for 1 hour at room temperature before development with Optiblot ECL reagent (ab133456) and imaging.

All lanes:

Western blot - Anti-CD27 antibody [EPR8568] (<a href='/en-us/products/primary-antibodies/cd27-antibody-epr8568-ab133761'>ab133761</a>) at 1/1000 dilution

Lane 1:

Wild-type Raji cell lysate at 20 µg

Lane 2:

CD27 knockout Raji cell lysate at 20 µg

Lane 2:

Western blot - Human CD27A knockout Raji cell line (<a href='/en-us/products/cell-lines/human-cd27a-knockout-raji-cell-line-ab274910'>ab274910</a>)

Lane 3:

Ramos cell lysate at 20 µg

Lane 4:

Human Brain tissue lysate at 20 µg

Predicted band size: 29 kDa

Observed band size: 50 kDa

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Exposure time: 2min

Western blot - Human CD27A knockout Raji cell lysate (AB281360)
  • WB

Lab

Western blot - Human CD27A knockout Raji cell lysate (AB281360)

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

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

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

Lane 4 : Human brain tissue lysate 20 ug

Lanes 1 - 4 : Merged signal (red and green). Green - ab131254 observed at 35 kDa. Red - 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 cell line ab274910 (knockout cell lysate ab281360). 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 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.

All lanes:

Western blot - Anti-CD27 antibody [EPR8569] (<a href='/en-us/products/primary-antibodies/cd27-antibody-epr8569-ab131254'>ab131254</a>) 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 2:

Western blot - Human CD27A knockout Raji cell line (<a href='/en-us/products/cell-lines/human-cd27a-knockout-raji-cell-line-ab274910'>ab274910</a>)

Lane 3:

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

Lane 4:

Human brain tissue lysate at 20 µg

Predicted band size: 29 kDa

Observed band size: 35 kDa

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Next Generation Sequencing - Human CD27A knockout Raji cell lysate (AB281360)
  • NGS

Supplier Data

Next Generation Sequencing - Human CD27A knockout Raji cell lysate (AB281360)

Knockout achieved by CRISPR/Cas9; X = 10 bp deletion; Frameshift : 100%

Key facts

Cell type

Raji

Species or organism

Human

Tissue

Lymphatic

Knockout validation

Next Generation Sequencing,Western blot

Mutation description

Knockout achieved by CRISPR/Cas9 X = 10 bp deletion Frameshift: 100%

Disease

Lymphoma

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Complementary Products

Primary Antibodies

AB236155

Anti-CD27 antibody [EPR20585] - BSA and Azide free

RabMAb|Recombinant

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD27 antibody [EPR20585] - BSA and Azide free (AB236155)

  • 0
  • 0
Assay Kits

AB289648

Human CD27 ELISA Kit

Recombinant|SimpleStep

ELISA - Human CD27 ELISA Kit (AB289648)

  • 0
  • 0
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Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Reactivity", "Dilution Info", "Notes"] }, "values": { "WB": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"<p></p>" } } }
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Product details

Knockout cell lysate achieved by CRISPR/Cas9.

REACH authorisation
Abcam has not and does not intend to apply for the REACH Authorisation of customers' uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.

Lysate preparation: Our lysates are made using RIPA buffer to which we add a protease inhibitor cocktail and phosphatase inhibitor cocktail (ratio: 300:100:10). This means that the protein of interest is denatured. If you require a native form of the protein please use the live cell version. Please refer to our lysis protocol for further details on how our lysates are prepared.

User storage instructions: Lyophilizate may be stored at 4°C. After reconstitution, store at -20°C for short-term storage or -80°C for long-term storage.

This product is subject to limited use licenses from The Broad Institute and ERS Genomics Limited, and is developed with patented technology. For full details of the limited use licenses and relevant patents please refer to our limited use license and patent pages.

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What's included?

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Properties and storage information

Gene name
CD27
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Next Generation Sequencing, Western blot
Shipped at conditions
Ambient - Can Ship with Ice
Appropriate short-term storage conditions
-20°C
Appropriate long-term storage conditions
-20°C
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Supplementary information

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

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.

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.
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Quality control

STR analysis

CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX

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Cell culture

Biosafety level

EU: 2 US: 2

Adherent/suspension

Suspension

Gender

Male

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Product protocols

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Product promise

We are committed to supporting your work with high-quality reagents, and we're here for you every step of the way. In the unlikely event that one of our products does not perform as expected, you're protected by our Product Promise.
For full details, please see our Terms & Conditions

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

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