Recombinant Anti-Neurofibromin antibody [EPR22989-68] (ab238142)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR22989-68] to Neurofibromin
- Suitable for: WB
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-Neurofibromin antibody [EPR22989-68]
See all Neurofibromin primary antibodies -
Description
Rabbit monoclonal [EPR22989-68] to Neurofibromin -
Host species
Rabbit -
Tested applications
Suitable for: WBmore details
Unsuitable for: Flow Cyt,ICC/IF,IHC-P or IP -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: HAP1, HeLa, Mouse brain, Rat brain, NIH/3T3, MEF and PC-12 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
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
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. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR22989-68 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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KO cell lines
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KO cell lysates
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab238142 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application | Abreviews | Notes |
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WB |
1/1000. Predicted molecular weight: 319 kDa.
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Notes |
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WB
1/1000. Predicted molecular weight: 319 kDa. |
Target
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Function
Stimulates the GTPase activity of Ras. NF1 shows greater affinity for Ras GAP, but lower specific activity. May be a regulator of Ras activity. -
Involvement in disease
Defects in NF1 are the cause of neurofibromatosis type 1 (NF1) [MIM:162200]; also known as von Recklinghausen syndrome. A disease characterized by patches of skin pigmentation (cafe-au-lait spots), Lisch nodules of the iris, tumors in the peripheral nervous system and fibromatous skin tumors. Individuals with the disorder have increased susceptibility to the development of benign and malignant tumors.
Defects in NF1 are a cause of juvenile myelomonocytic leukemia (JMML) [MIM:607785]. JMML is a pediatric myelodysplastic syndrome that constitutes approximately 30% of childhood cases of myelodysplastic syndrome (MDS) and 2% of leukemia. Germline mutations of NF1 account for the association of JMML with type 1 neurofibromatosis (NF1).
Defects in NF1 are the cause of Watson syndrome (WS) [MIM:193520]. WS is characterized by the presence of pulmonary stenosis, cafe-au-lait spots, and mental retardation. WS is considered as an atypical form of NF1.
Defects in NF1 are a cause of familial spinal neurofibromatosis (FSNF) [MIM:162210]. Familial spinal NF is considered to be an alternative form of neurofibromatosis, showing multiple spinal tumors.
Defects in NF1 are a cause of neurofibromatosis-Noonan syndrome (NFNS) [MIM:601321]. NFNS is characterized by manifestations of both NF1 and Noonan syndrome (NS). NS is a disorder characterized by dysmorphic facial features, short stature, hypertelorism, cardiac anomalies, deafness, motor delay, and a bleeding diathesis.
Defects in NF1 may be a cause of colorectal cancer (CRC) [MIM:114500]. -
Sequence similarities
Contains 1 CRAL-TRIO domain.
Contains 1 Ras-GAP domain. - Information by UniProt
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Database links
- Entrez Gene: 4763 Human
- Entrez Gene: 18015 Mouse
- Entrez Gene: 24592 Rat
- Omim: 613113 Human
- SwissProt: P21359 Human
- SwissProt: Q04690 Mouse
- SwissProt: P97526 Rat
- Unigene: 113577 Human
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Alternative names
- DKFZp686J1293 antibody
- FLJ21220 antibody
- Neurofibromatosis Noonan syndrome antibody
see all
Images
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All lanes : Anti-Neurofibromin antibody [EPR22989-68] (ab238142) at 1/1000 dilution
Lane 1 : Wild-type MCF7 cell lysate
Lane 2 : NF1 knockout MCF7 cell lysate
Lane 3 : Wild Type HeLa ab255929 cell lysate
Lane 4 : NF1 knockout Hela ab258533 cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 319 kDa
Observed band size: 180-270 kDa why is the actual band size different from the predicted?Anti-NF1 antibody [EPR22989-68] (ab238142) staining at 1/1000 dilution, shown in green; Mouse anti-CANX [CANX/1543] (ab238078) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab238142 was shown to bind specifically to NF1. A band was observed at 180-270 kDa in wild-type MCF7 cell lysates with no signal observed at this size in NF1 knockout cell line. To generate this image, wild-type and NF1 knockout MCF7 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.
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All lanes : Anti-Neurofibromin antibody [EPR22989-68] (ab238142) at 1/1000 dilution
Lane 1 : Wild-type HeLa cell lysate
Lane 2 : NFIB knockout HeLa cell lysate
Lane 3 : HAP1 cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 319 kDaLanes 1-3: Merged signal (red and green). Green - ab238142 observed at 319 kDa. Red - loading control ab7291 observed at 50 kDa.
ab238142 Recombinant Anti-NF1 antibody [EPR22989-68] was shown to specifically react with NFIB in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab264725 (knockout cell lysate ab258533) was used. Wild-type and NF1 knockout samples were subjected to SDS-PAGE. ab238142 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
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All lanes : Anti-Neurofibromin antibody [EPR22989-68] (ab238142) at 1/1000 dilution
Lane 1 : Wild-type HAP1 whole cell lysate
Lane 2 : Neurofibromin knockout HAP1 whole cell lysate
Lane 3 : HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution
Predicted band size: 319 kDa
Observed band size: 250,300 kDa why is the actual band size different from the predicted?ab238142 was shown to specifically react with Neurofibromin in wild-type HAP1 cells as signal was lost in Neurofibromin knockout cells. Wild-type and Neurofibromin knockout samples were subjected to SDS-PAGE. ab238142 and ab129002 (Rabbit anti-Vinculin loading control) were incubated 1 hour at room temperature at 1/1000 dilution and 1/5000 dilution respectively. Blots were developed with Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) secondary antibody at 1/100,000 dilution for 1 hour at room temperature before imaging. The blot was developed on a BIO-RAD® ChemiDoc™ MP instrument using the ECL technique. Lysate should be made freshly and used in WB immediately to minimize protein degradation. Degraded fragment(250 KD) observed is consistent with what has been described in the literature (PMID: 30131853, PMID: 30408279).
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
Exposure time: 26 seconds
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All lanes : Anti-Neurofibromin antibody [EPR22989-68] (ab238142) at 1/1000 dilution
Lane 1 : Mouse brain lysate
Lane 2 : Rat brain lysate
Lane 3 : NIH/3T3 (mouse embryonic fibroblast) whole cell lysate
Lane 4 : MEF (mouse embryonic fibroblast (immortalized)) whole cell lysate
Lane 5 : PC-12 (rat adrenal gland heochromocytoma) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 319 kDa
Observed band size: 200,250,300 kDa why is the actual band size different from the predicted?Degraded fragments (200-250 KD) observed is consistent with what has been described in the literature (PMID: 30131853, PMID: 30408279).
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
Exposure time: Lane 1:3 minutes; Lane 2:26 seconds; Lanes 3-5: 3 minutes.
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (0)
ab238142 has not yet been referenced specifically in any publications.