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AB273812

Human CD86 knockout Raji cell lysate

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CD86 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 = 1 bp insertion; Frameshift: 99%.

View Alternative Names

Activation B7-2 antigen, Activation B7-2 antigen 3, B-lymphocyte activation antigen B7-2, B-lymphocyte activation antigen B7-2 2, B7, B7-2, B7.2, B70, B72 antigen, BU63, CD28 antigen ligand 2, CD28 antigen ligand 2 2, CD28LG2, CD86 antigen, CD86 antigen (CD28 antigen ligand 2, B7-2 antigen), CD86 antigen (CD28 antigen ligand 2, B7-2 antigen) 1, 2, CD86 molecule, CD86_HUMAN, CLS1, CTLA-4 counter-receptor B7.2, CTLA-4 counter-receptor B7.2 2, CTLA-4 counter-receptor B7.2 2, 3, Cd28l2, ETC-1, Early T-cell costimulatory molecule 1, FUN-1, LAB72, Ly-58, MB7, MB7-2, MGC34413, Membrane glycoprotein, T lymphocyte activation antigen CD86 precursor, T-lymphocyte activation antigen CD86, TS/A-2

4 Images
Western blot - Human CD86 knockout Raji cell lysate (AB273812)
  • WB

Lab

Western blot - Human CD86 knockout Raji cell lysate (AB273812)

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

Lane 2 : CD86 knockout Raji (Human Burkitt's lymphoma cell line) whole cell lysate

Lanes 1 - 2 : Merged signal (red and green). Green - ab269587 observed at 70 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55kDa.

ab269587 was shown to react with CD86 in wild-type Raji cells in western blot with loss of signal observed in CD86 knockout cell line ab273858 (knockout cell lysate ab273812). Wild-type and CD86 knockout Raji cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with ab269587 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-CD86 antibody [EP1158-37] (<a href='/en-us/products/primary-antibodies/cd86-antibody-ep1158-37-ab269587'>ab269587</a>) at 1/1000 dilution

Lane 1:

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

Lane 2:

CD86 knockout Raji (Human Burkitt's lymphoma cell line) whole cell lysate at 20 µg

Lane 2:

Western blot - Human CD86 knockout Raji cell line (<a href='/en-us/products/cell-lines/human-cd86-knockout-raji-cell-line-ab273858'>ab273858</a>)

Predicted band size: 38 kDa

Observed band size: 70 kDa

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Western blot - Human CD86 knockout Raji cell lysate (AB273812)
  • WB

Lab

Western blot - Human CD86 knockout Raji cell lysate (AB273812)

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

Lane 2 : CD86 knockout Raji (Human Burkitt's lymphoma cell line) whole cell lysate

Lanes 1 - 2 : Merged signal (red and green). Green - ab239075 observed at 70 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55kDa.

ab239075 was shown to react with CD86 in wild-type Raji cells in western blot with loss of signal observed in CD86 knockout cell line ab273858 (knockout cell lysate ab273812). Wild-type and CD86 knockout Raji cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with ab239075 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-CD86 antibody [EPR21962] (<a href='/en-us/products/primary-antibodies/cd86-antibody-epr21962-ab239075'>ab239075</a>) at 1/1000 dilution

Lane 1:

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

Lane 2:

CD86 knockout Raji (Human Burkitt's lymphoma cell line) whole cell lysate at 20 µg

Lane 2:

Western blot - Human CD86 knockout Raji cell line (<a href='/en-us/products/cell-lines/human-cd86-knockout-raji-cell-line-ab273858'>ab273858</a>)

Predicted band size: 38 kDa

Observed band size: 70 kDa

false

Western blot - Human CD86 knockout Raji cell lysate (AB273812)
  • WB

Lab

Western blot - Human CD86 knockout Raji cell lysate (AB273812)

Lane 1 : Wild-type Raji (Human Burkitt's lymphoma cell line) whole cell lysate
Lane 2 : CD86 knockout Raji (Human Burkitt's lymphoma cell line) whole cell lysate
Lane 3 : Ramos whole cell lysate
Lane 4 : Human liver cell lysate

Lanes 1 - 4 : Merged signal (red and green). Green - ab220188 observed at 70 kDa. Red - loading control ab52866 (Rabbit anti-alpha Tubulin antibody [EP1332Y]) observed at 55 kDa.

ab220188 was shown to react with CD86 in wild-type Raji cells in Western blot with loss of signal observed in CD86 knockout cell line ab273812 (knockout cell lysate ab273858). Wild-type Raji and CD86 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with ab220188 and ab52866 (Rabbit anti-alpha Tubulin antibody [EP1332Y]) overnight at 4 °C at 0.5 μg/ml and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed (ab216777) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.

All lanes:

Western blot - Anti-CD86 antibody [C86/1146] (<a href='/en-us/products/primary-antibodies/cd86-antibody-c86-1146-ab220188'>ab220188</a>) at 0.5 µg/mL

Lane 1:

Wild-type Raji cell lysate at 20 µg

Lane 2:

CD86 knockout Raji cell lysate at 20 µg

Lane 2:

Western blot - Human CD86 knockout Raji cell line (<a href='/en-us/products/cell-lines/human-cd86-knockout-raji-cell-line-ab273858'>ab273858</a>)

Lane 3:

Ramos cell lysate at 20 µg

Lane 4:

Human Liver tissue lysate at 20 µg

Predicted band size: 38 kDa

Observed band size: 70 kDa

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Next Generation Sequencing - Human CD86 knockout Raji cell lysate (AB273812)
  • NGS

Supplier Data

Next Generation Sequencing - Human CD86 knockout Raji cell lysate (AB273812)

Knockout achieved by CRISPR/Cas9; X = 1 bp insertion; Frameshift : 99%

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 = 1 bp insertion; Frameshift: 99%

Disease

Lymphoma

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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, ERS Genomics Limited and Sigma-Aldrich Co. LLC, and is developed with patented technology. For full details of the licenses and 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
CD86
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.

CD86 also known as B7-2 is a protein involved in the regulation of the immune response. It has an approximate mass of 70 kDa and is expressed on antigen-presenting cells like dendritic cells monocytes and macrophages. Notably CD86 is present on macrophages including those in tissues such as skin and lymphoid organs. Expressed on these cells CD86 serves as a vital mediator in the co-stimulatory signals necessary for T cell activation and survival.
Biological function summary

CD86 plays a significant role in the immune system by providing secondary signals for T cell activation and differentiation. It is a part of the B7 protein family and forms a complex with CD28 and CTLA-4 on T cells. When CD86 binds to CD28 it sends positive co-stimulatory signals which promote T cell proliferation and cytokine production. On the other hand interaction with CTLA-4 transmits an inhibitory signal which reduces immune response. This dual interaction helps to balance immune activation and tolerance.

Pathways

CD86 takes part in important immune-related signaling pathways particularly the T cell receptor signaling pathway and the PI3K-Akt signaling pathway. Both pathways are fundamental for initiating immune responses. CD86's interaction with CD28 activates downstream signaling cascades including PI3K-Akt which is important for cell survival and growth. Additionally CD86 collaborates with other proteins such as CD80 another co-stimulatory molecule to amplify T cell activation within these pathways.

CD86 is associated with autoimmune diseases and transplant rejection. In autoimmune diseases like rheumatoid arthritis the overexpression or dysregulation of CD86 can lead to excessive T cell activation causing immune system attacks on the body's own tissues. Similarly in transplant rejection CD86 may contribute by enhancing immune response against transplanted organs. The engagement between CD86 and CD28 is a critical factor in these conditions and therapies targeting this interaction are under exploration to mitigate the immune response.
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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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Primary Antibodies

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