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AB273858

Human CD86 knockout Raji cell line

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CD86 KO cell line available to order. KO validated by Next Generation Sequencing, Western blot. Free of charge wild type control available. Knockout achieved by CRISPR/Cas9; X = 1 bp insertion; Frameshift: 99%. To order both knockout and wild-type control cells: select '2 x 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.

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

5 Images
Western blot - Human CD86 knockout Raji cell line (AB273858)
  • WB

Lab

Western blot - Human CD86 knockout Raji cell line (AB273858)

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 sample. 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° 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 (ab273858)

Predicted band size: 38 kDa

Observed band size: 70 kDa

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Western blot - Human CD86 knockout Raji cell line (AB273858)
  • WB

Lab

Western blot - Human CD86 knockout Raji cell line (AB273858)

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 sample. 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° 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 (ab273858)

Predicted band size: 38 kDa

Observed band size: 70 kDa

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Western blot - Human CD86 knockout Raji cell line (AB273858)
  • WB

Lab

Western blot - Human CD86 knockout Raji cell line (AB273858)

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 ° 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 (ab273858)

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 line (AB273858)
  • NGS

Supplier Data

Next Generation Sequencing - Human CD86 knockout Raji cell line (AB273858)

1 bp insertion after Leu18 of the WT protein

Next Generation Sequencing - Human CD86 knockout Raji cell line (AB273858)
  • NGS

Supplier Data

Next Generation Sequencing - Human CD86 knockout Raji cell line (AB273858)

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

Key facts

Cell type

Raji

Species or organism

Human

Tissue

Lymphatic

Form

Liquid

form

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": { "NGS": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"<p></p>" }, "WB": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"<p></p>" } } }
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Product details

We will provide viable cells that proliferate on revival.

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?

{ "values": { "2x1000000Cellsvial": { "sellingSize": "2 x 1000000 Cells/vial", "publicAssetCode":"ab273858-2x1000000Cells_vial", "assetComponentDetails": [ { "size":"1 x 1000000 Cells/vial", "name":"ab271145 Human wild-type Raji cell line", "number":"AB273858-CMP02" }, { "size":"1 x 1000000 Cells/vial", "name":"ab273858 Human CD86 knockout Raji cell line", "number":"AB273858-CMP01" } ] }, "1000000Cellsvial": { "sellingSize": "1000000 Cells/vial", "publicAssetCode":"ab273858-1000000Cells_vial", "assetComponentDetails": [ { "size":"1 x 1000000 Cells/vial", "name":"ab273858 Human CD86 knockout Raji cell line", "number":"AB273858-CMP01", "productcode":"" } ] } } }
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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
Dry Ice
Appropriate short-term storage conditions
-196°C
Appropriate long-term storage conditions
-196°C
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Handling procedures

Initial handling guidelines

Upon arrival, the vial should be stored in liquid nitrogen vapor phase and not at -80°C. Storage at -80°C may result in loss of viability.

1. Thaw the vial in 37°C water for bath approximately 1-2 minutes.
2. Transfer the cell suspension (0.8 mL) to a 15 mL/50 mL conical sterile polypropylene centrifuge tube containing 8.4 mL pre-warmed culture medium, wash vial with an additional 0.8 mL culture medium (total volume 10 mL) to collect remaining cells, and centrifuge at 201 x g (rcf) for 5 minutes at room temperature. 10 mL represents minimum recommended dilution. 20 mL represents maximum recommended dilution.
3. Resuspend the cell pellet in 5 mL pre-warmed culture medium and count using a haemocytometer or alternative cell counting method. Based on cell count, seed cells in an appropriate cell culture flask at a density of 4x105 - 5x105 cells/mL(for initial passages it is recomended to culture the cells in the higher range of recomended seeding density). Seeding density is given as a guide only and should be scaled to align with individual lab schedules.
4. Incubate the culture at 37°C incubator with 5% CO2. Cultures should be monitored daily.

Subculture guidelines
  • All seeding densities should be based on cell counts gained by established methods.
  • A guide seeding density of 4x105 cells/mL is recommended.
  • A maximum of 3x106 viable cells/mL is obtainable.
Culture medium

RPMI + 10% FBS

Cryopreservation medium

Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.

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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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Target data

See full target information CD86
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Western blot - Human CD80 knockout Raji cell line (AB273873)

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