Anti-ITCH/AIP4 antibody [32/Itch] (ab280211)
![Anti-ITCH/AIP4 antibody [32/Itch] (ab280211)](https://www.abcam.com/ps/products/280/ab280211/Images/ab280211-456779-anti-itchaip4-antibody-32itch-western-blot-hela.jpg "Anti-ITCH/AIP4 antibody [32/Itch] (ab280211)")
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Mouse monoclonal [32/Itch] to ITCH/AIP4
- Suitable for: WB
- Knockout validated
- Reacts with: Mouse, Rat, Human
Overview
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Product name
Anti-ITCH/AIP4 antibody [32/Itch]
See all ITCH/AIP4 primary antibodies -
Description
Mouse monoclonal [32/Itch] to ITCH/AIP4 -
Host species
Mouse -
Tested applications
Suitable for: WBmore details
Unsuitable for: ICC -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: Hela, Ramos, Raji, C6, PC-12, NIH/3T3, K562 whole cell lysates.
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
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
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading... -
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
32/Itch -
Isotype
IgG1 -
Research areas
Images
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Western blot - Anti-ITCH/AIP4 antibody [32/Itch] (ab280211)All lanes : Anti-ITCH/AIP4 antibody [32/Itch] (ab280211) at 1/1000 dilution
Lane 1 : HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate
Lane 2 : Ramos (human Burkitt's lymphoma B lymphocyte), whole cell lysate
Lane 3 : Raji (human Burkitt's lymphoma B lymphocyte), whole cell lysate
Lane 4 : C6 (rat glial tumor glial cell), whole cell lysate
Lane 5 : PC-12 (rat adrenal gland pheochromocytoma), whole cell lysate
Lane 6 : NIH/3T3 (mouse embryonic fibroblast), whole cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Peroxidase-Conjugated Goat anti-Mouse IgG (H+L) at 1/10000 dilution
Predicted band size: 103 kDa
Observed band size: 120 kDa why is the actual band size different from the predicted?Blocking and Diluting buffer and concentration: 5% NFDM/TBST
The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID:11782481).
The band around 75kDa is the degradation form. 50kDa band is the non-specific band according to KO validation.
Exposure time Lane 1-2/4-5: 3 minutes Lane 3/6: 1 minute
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![Western blot - Anti-ITCH/AIP4 antibody [32/Itch] (ab280211)](https://www.abcam.com/ps/products/280/ab280211/Images/ab280211-456781-anti-itchaip4-antibody-32itch-western-blot-wildtype-hela.jpg)
Western blot - Anti-ITCH/AIP4 antibody [32/Itch] (ab280211)All lanes : Anti-ITCH/AIP4 antibody [32/Itch] (ab280211) at 1/1000 dilution
Lane 1 : Wild-type HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate
Lane 2 : ITCH knockout HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate
Lane 3 : K562 (human chronic myelogenous leukemia lymphoblast), whole cell lysate
Lane 4 : Raji (human Burkitt's lymphoma B lymphocyte), whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Mouse IgG H&L (IRDye® 800CW) (ab216772) and Goat Anti-Rabbit IgG H&L (IRDye® 680RD) (ab216777) at 1/10000 dilution
Predicted band size: 103 kDa
Observed band size: 120 kDa why is the actual band size different from the predicted?Blocking and Diluting buffer and concentration: Intercept® (TBS) Blocking Buffer diluted with an equal volume of 0.1% TBS
Lanes 1 - 4: Merged signal (red and green). Green - ab280211 observed at 120 kDa. Red - loading control ab181602 (Rabbit monoclonal [EPR16891] to GAPDH) observed at 36 kDa.
Lanes 1-2: ab280211 Anti-ITCH antibody was shown to react with ITCH in HeLa cells in Western blot. Loss of signal was observed when ITCH knockout sample was used. Wild-type and ITCH knockout samples were subjected to SDS-PAGE.
ab280211 and Anti-GAPDH antibody [EPR16891] (ab181602) were incubated at 4? overnight at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 680CW) (ab216777) and Goat anti-Mouse IgG H&L (IRDye® 800RD) (ab216772) secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.
The band around 75kDa is the degradation form. 50kDa band is the non-specific band according to KO validation.
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![Anti-ITCH/AIP4 antibody [32/Itch] (ab280211)](https://www.abcam.com/ps/products/280/ab280211/Images/ab280211-1-Benefits-of-Recombinant-antibodies.jpg)
Anti-ITCH/AIP4 antibody [32/Itch] (ab280211)
