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AB275529

Human CD74 knockout Raji cell lysate

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CD74 KO cell lysate available now. KO validated. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, Homozygous: 13 bp deletion in exon 2.

View Alternative Names

CD74 antigen, CD74 antigen (invariant polypeptide of major histocompatibility complex, class II antigen-associated), CD74 molecule, CD74 molecule, major histocompatibility complex, class II invariant chain, DHLAG, Gamma chain of class II antigens, HG2A_HUMAN, HLA class II histocompatibility antigen gamma chain, HLA-DR antigens-associated invariant chain, HLA-DR-gamma, HLADG, Ia antigen-associated invariant chain, Ia associated invariant chain, Ia gamma, Ii, Invariant polypeptide of major histocompatibility complex class II antigen associated, MHC HLA-DR gamma chain, Major histocompatibility complex class II invariant chain, P35, Protein 41, la-gamma, p33

6 Images
Western blot - Human CD74 knockout Raji cell lysate (AB275529)
  • WB

Lab

Western blot - Human CD74 knockout Raji cell lysate (AB275529)

Lane 1 : Wild-type Raji cell lysate 30 ug
Lane 2 : CD74 knockout Raji cell lysate 30 ug
Lane 3 : Jurkat cell lysate 30 ug
Lane 4 : HepG2 cell lysate 30 ug
Lanes 1 - 4 : Merged signal (red and green). Green - ab270265 observed at 35 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55 kDa.
ab270265 was shown to react with CD74 in western blot. The band observed in the knockout lysate lane below 35kDa may represent truncated forms and cleaved fragments. This has not been investigated further. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with ab270265 and ab7291 (Mouse anti-Alpha Tubulin [DM1A]) overnight at 4 °C at 1 ug/ml 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 h at room temperature before imaging.

All lanes:

Western blot - Anti-CD74 antibody [CLIP/3127R] (<a href='/en-us/products/primary-antibodies/cd74-antibody-clip-3127r-ab270265'>ab270265</a>) at 1 µg/mL

Lane 1:

Wild-type Raji cell lysate at 30 µg

Lane 2:

CD74 knockout Raji cell lysate at 30 µg

Lane 2:

Western blot - Human CD74 knockout Raji cell line (<a href='/en-us/products/cell-lines/human-cd74-knockout-raji-cell-line-ab273378'>ab273378</a>)

Lane 3:

Jurkat cell lysate at 30 µg

Lane 4:

HepG2 cell lysate at 30 µg

Predicted band size: 34 kDa

Observed band size: 35 kDa

false

Western blot - Human CD74 knockout Raji cell lysate (AB275529)
  • WB

Lab

Western blot - Human CD74 knockout Raji cell lysate (AB275529)

Lane 1 : Wild-type Raji cell lysate 30 ug
Lane 2 : CD74 knockout Raji cell lysate 30 ug
Lane 3 : Jurkat cell lysate 30 ug
Lane 4 : HepG2 cell lysate 30 ug
Lanes 1 - 4 : Merged signal (red and green). Green - ab22603 observed at 35 kDa. Red - loading control ab52866 (Rabbit anti-alpha Tubulin antibody [EP1332Y]) observed at 55 kDa.
ab22603 was shown to react with CD74 in western blot. The band observed in the knockout lysate lane below 35kDa may represent truncated forms and cleaved fragments. This has not been investigated further. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with ab22603 and ab52866 (Rabbit anti-alpha Tubulin antibody [EP1332Y]) overnight at 4 °C at 1 ug/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-CD74 antibody [PIN.1] (<a href='/en-us/products/primary-antibodies/cd74-antibody-pin1-ab22603'>ab22603</a>) at 1 µg/mL

Lane 1:

Wild-type Raji cell lysate at 30 µg

Lane 2:

Western blot - Human CD74 knockout Raji cell lysate (ab275529)

Lane 2:

CD74 knockout Raji cell lysate at 30 µg

Lane 2:

Western blot - Human CD74 knockout Raji cell line (<a href='/en-us/products/cell-lines/human-cd74-knockout-raji-cell-line-ab273378'>ab273378</a>)

Lane 3:

Jurkat cell lysate at 30 µg

Lane 4:

HepG2 cell lysate at 30 µg

Predicted band size: 34 kDa

Observed band size: 35 kDa

false

Western blot - Human CD74 knockout Raji cell lysate (AB275529)
  • WB

Lab

Western blot - Human CD74 knockout Raji cell lysate (AB275529)

Lane 1 : Wild-type Raji cell lysate 30 ug
Lane 2 : CD74 knockout Raji cell lysate 30 ug
Lane 3 : Jurkat cell lysate 30 ug
Lane 4 : HepG2 cell lysate 30 ug
Lanes 1 - 4 : Merged signal (red and green). Green - ab64772 observed at 35 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55 kDa.
ab64772 was shown to react with CD74 in western blot. The band observed in the knockout lysate lane below 35kDa may represent truncated forms and cleaved fragments. This has not been investigated further. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with ab64772 and ab7291 (Mouse anti-Alpha Tubulin [DM1A]) overnight at 4 °C at 1 ug/ml 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 h at room temperature before imaging.

All lanes:

Western blot - Anti-CD74 antibody (<a href='/en-us/products/primary-antibodies/cd74-antibody-ab64772'>ab64772</a>) at 1 µg/mL

Lane 1:

Wild-type Raji cell lysate at 30 µg

Lane 2:

CD74 knockout Raji cell lysate at 30 µg

Lane 2:

Western blot - Human CD74 knockout Raji cell lysate (ab275529)

Lane 2:

Western blot - Human CD74 knockout Raji cell line (<a href='/en-us/products/cell-lines/human-cd74-knockout-raji-cell-line-ab273378'>ab273378</a>)

Lane 3:

Jurkat cell lysate at 30 µg

Lane 4:

HepG2 cell lysate at 30 µg

Predicted band size: 34 kDa

Observed band size: 35 kDa

false

Western blot - Human CD74 knockout Raji cell lysate (AB275529)
  • WB

Lab

Western blot - Human CD74 knockout Raji cell lysate (AB275529)

Lane 1 : Wild-type Raji cell lysate 30 ug
Lane 2 : CD74 knockout Raji cell lysate 30 ug
Lane 3 : Jurkat cell lysate 30 ug
Lane 4 : HepG2 cell lysate 30 ug
Lanes 1 - 4 : Merged signal (red and green). Green - ab108393 observed at 35 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55 kDa.
ab108393 was shown to react with CD74 in western blot. The band observed in the knockout lysate lane below 35kDa may represent truncated forms and cleaved fragments. This has not been investigated further. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with ab108393 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 h at room temperature before imaging.

All lanes:

Western blot - Anti-CD74 antibody [EPR4064] (<a href='/en-us/products/primary-antibodies/cd74-antibody-epr4064-ab108393'>ab108393</a>) at 1/1000 dilution

Lane 1:

Wild-type Raji cell lysate at 30 µg

Lane 2:

Western blot - Human CD74 knockout Raji cell lysate (ab275529)

Lane 2:

CD74 knockout Raji cell lysate at 30 µg

Lane 2:

Western blot - Human CD74 knockout Raji cell line (<a href='/en-us/products/cell-lines/human-cd74-knockout-raji-cell-line-ab273378'>ab273378</a>)

Lane 3:

Jurkat cell lysate at 30 µg

Lane 4:

HepG2 cell lysate at 30 µg

Predicted band size: 34 kDa

Observed band size: 35 kDa

false

Western blot - Human CD74 knockout Raji cell lysate (AB275529)
  • WB

Lab

Western blot - Human CD74 knockout Raji cell lysate (AB275529)

Lane 1 : Wild-type Raji cell lysate 30 ug
Lane 2 : CD74 knockout Raji cell lysate 30 ug
Lane 3 : Jurkat cell lysate 30 ug
Lane 4 : HepG2 cell lysate 30 ug
Lanes 1 - 4 : Merged signal (red and green). Green - ab9514 observed at 35 kDa. Red - loading control ab52866 (Rabbit anti-alpha Tubulin antibody [EP1332Y]) observed at 55 kDa.
ab9514 was shown to react with CD74 in western blot. The band observed in the knockout lysate lane below 35kDa may represent truncated forms and cleaved fragments. This has not been investigated further. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with ab9514 and ab52866 (Rabbit anti-alpha Tubulin antibody [EP1332Y]) overnight at 4 °C at 5 ug/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-CD74 antibody [LN2] (<a href='/en-us/products/primary-antibodies/cd74-antibody-ln2-ab9514'>ab9514</a>) at 5 µg/mL

Lane 1:

Wild-type Raji cell lysate at 30 µg

Lane 2:

CD74 knockout Raji cell lysate at 30 µg

Lane 2:

Western blot - Human CD74 knockout Raji cell lysate (ab275529)

Lane 2:

Western blot - Human CD74 knockout Raji cell line (<a href='/en-us/products/cell-lines/human-cd74-knockout-raji-cell-line-ab273378'>ab273378</a>)

Lane 3:

Jurkat cell lysate at 30 µg

Lane 4:

HepG2 cell lysate at 30 µg

Predicted band size: 34 kDa

Observed band size: 35 kDa

false

Sanger Sequencing - Human CD74 knockout Raji cell lysate (AB275529)
  • Sanger seq

Supplier Data

Sanger Sequencing - Human CD74 knockout Raji cell lysate (AB275529)

Homozygous : 13 bp deletion in exon 2

Key facts

Cell type

Raji

Species or organism

Human

Tissue

Lymphatic

Knockout validation

Sanger Sequencing

Mutation description

Knockout achieved by using CRISPR/Cas9, Homozygous: 13 bp deletion in exon 2

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>Western blot data indicates that the CRISPR gene edit may have resulted in a truncation of the protein of interest. Please see data images.</p>" } } }
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Product details

Western blot data indicates that the CRISPR gene edit may have resulted in a truncation of the protein of interest. Please see data images.

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?

{ "values": { "1Kit": { "sellingSize": "1 Kit", "publicAssetCode":"ab275529-1Kit", "assetComponentDetails": [ { "size":"1 x 100 µg", "name":"Human CD74 knockout Raji cell lysate", "number":"AB275529-CMP01", "productcode":"" }, { "size":"1 x 100 µg", "name":"Human wild-type Raji cell lysate", "number":"AB275529-CMP02", "productcode":"" } ] } } }
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Properties and storage information

Gene name
CD74
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Sanger Sequencing
Zygosity
Homozygous
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.

CD74 also known as the invariant chain or li is a protein with a mass of about 33 kDa. It is widely expressed on the surface of immune cells such as B cells dendritic cells and macrophages. CD74 plays an important role in the immune system by acting as a chaperone for major histocompatibility complex class II (MHC II) molecules guiding their migration to endosomes where they encounter antigenic peptides. The CD74 protein also functions as a cell surface receptor for macrophage migration inhibitory factor (MIF) enhancing the immune response process.
Biological function summary

CD74 facilitates multiple immune regulatory processes. It forms a complex with the MHC II molecules aiding in their proper folding and peptide loading in the endoplasmic reticulum. This complex then travels through the Golgi apparatus towards the lysosomal compartments where antigens are presented to initiate adaptive immune responses. In addition CD74 is involved in signal transduction pathways that regulate cell proliferation and survival influenced by interactions with MIF and other molecules.

Pathways

The role of CD74 extends into both the antigen presentation and MIF signaling pathways. Within the antigen presentation pathway CD74's activity is tightly connected with MHC II and subsequently to CD4+ T cells facilitating the activation of these immune cells. The MIF signaling pathway involves CD74 interacting with CD44 a receptor that further propagates the signaling cascades essential for immune modulation and cell survival.

CD74 is linked to inflammatory and autoimmune conditions such as rheumatoid arthritis and systemic lupus erythematosus. Aberrant expression of CD74 influences the presentation of autoantigens and may contribute to the chronic inflammation seen in these disorders. Its connection to MIF which is a pro-inflammatory cytokine further implicates CD74 in these autoimmune scenarios by promoting persistent immune activation. Researchers are also exploring CD74 as a therapeutic target in certain cancers due to its influence on immune surveillance and tumor progression.
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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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Complementary Products

Primary Antibodies

AB64772

Anti-CD74 antibody

KO Validated

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD74 antibody (AB64772)

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Proteins & Peptides

AB315990

Recombinant Human CD74 Protein Standard (His tag)

Sandwich ELISA - Recombinant Human CD74 Protein Standard (His tag) (AB315990)

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Assay Kits

AB284618

Human CD74 ELISA Kit

Recombinant|SimpleStep

Sandwich ELISA - Human CD74 ELISA Kit (AB284618)

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Cell Lines & Lysates

AB273378

Human CD74 knockout Raji cell line

Advanced Validation

Western blot - Human CD74 knockout Raji cell line (AB273378)

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