Anti-CD74 antibody [LN2] (ab265584)
Key features and details
- Mouse monoclonal [LN2] to CD74
- Suitable for: IHC-P, WB, ICC/IF
- Reacts with: Human
- Isotype: IgG1
Overview
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Product name
Anti-CD74 antibody [LN2]
See all CD74 primary antibodies -
Description
Mouse monoclonal [LN2] to CD74 -
Host species
Mouse -
Tested Applications & Species
See all applications and species dataApplication Species ICC/IF HumanIHC-P HumanWB Human -
Immunogen
Tissue, cells or virus corresponding to Human CD74. (Human SU-DHL-4 lymphoma cells).
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Positive control
- IHC-P: Human colon carcinoma and lymph node tissues. ICC: Raji and Daudi cells. WB: Raji, HeLa, and Daudi whole cell lysates.
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General notes
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.
One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.
Learn more here.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
Storage buffer
pH: 6.8
Preservative: 0.09% Sodium azide
Constituent: PBS -
Concentration information loading...
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Purification notes
Purified from cell culture supernatant. -
Clonality
Monoclonal -
Clone number
LN2 -
Isotype
IgG1 -
Research areas
Images
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All lanes : Anti-CD74 antibody [LN2] (ab9514) at 5 µg/ml
Lane 1 : Wild-type Raji cell lysate
Lane 2 : CD74 knockout Raji cell lysate
Lane 3 : Jurkat cell lysate
Lane 4 : HepG2 cell lysate
Lysates/proteins at 30 µg per lane.
Performed under reducing conditions.
Predicted band size: 34 kDa
Observed band size: 35 kDa why is the actual band size different from the predicted?This data was developed using the same antibody clone in a different buffer formulation (ab9514).
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 CD74 knockout cell line ab273378 (knockout lysate ab275529) below 35 kDa is likely to represent a truncated form. 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 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.
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Formalin-fixed, paraffin-embedded human colon carcinoma tissue stained for CD74 using ab265584 at 1/100 dilution in immunohistochemical analysis. The secondary antibody was an HRP-conjugated goat anti-mouse IgG. Substrate: DAB
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Formalin-fixed, paraffin-embedded human lymph node tissue stained for CD74 using ab265584 at 1/100 dilution in immunohistochemical analysis. The secondary antibody was an HRP-conjugated goat anti-mouse IgG. Substrate: DAB
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Formalin-fixed, paraffin-embedded Raji cells stained for CD74 using ab265584 at 1/100 dilution in immunocytochemical analysis. The secondary antibody was an HRP-conjugated goat anti-mouse IgG. Substrate: DAB
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Formalin-fixed, paraffin-embedded Daudi cells stained for CD74 using ab265584 at 1/100 dilution in immunocytochemical analysis. The secondary antibody was an HRP-conjugated goat anti-mouse IgG. Substrate: DAB
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All lanes : Anti-CD74 antibody [LN2] (ab265584) at 1/1000 dilution
Lane 1 : HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate
Lane 2 : Raji (human Burkitt's lymphoma cell line) whole cell lysate
Lane 3 : GaMG whole cell lysate
Lane 4 : HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lane 5 : SR whole cell lysate
Lane 6 : Jurkat (human T cell leukemia cell line from peripheral blood) whole cell lysate
Lane 7 : Daudi (human Burkitt's lymphoma cell line) whole cell lysate
Lane 8 : KG-1 whole cell lysate
Lysates/proteins at 50 µg per lane.
Secondary
All lanes : HRP-conjugated goat anti-mouse IgG
Developed using the ECL technique.
Predicted band size: 34 kDa
Exposure time: 30 seconds