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V-Crk Sarcoma Virus CT10 Oncogene Homolog (Avian) (CRK) (AA 102-304) antibody

Details for Product No. ABIN967702
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Antigen
Synonyms
CG1587, CRK, D-CRK, D-Crk, DCrk, Dcrk, Dmel\\CG1587, crk, dCRK, dCrk, fb34h10, fc05a01, fi19g05, wu:fb34h10, wu:fc05a01, wu:fc54a04, wu:fi19g05, zgc:100936, CRKII, p38, Crko, crk2, crkII, v-crk, P38C- ... show more
CG1587, CRK, D-CRK, D-Crk, DCrk, Dcrk, Dmel\\CG1587, crk, dCRK, dCrk, fb34h10, fc05a01, fi19g05, wu:fb34h10, wu:fc05a01, wu:fc54a04, wu:fi19g05, zgc:100936, CRKII, p38, Crko, crk2, crkII, v-crk, P38C-CRK, Crk-I, Crk-II, Crk-III, Crk3, CrkIII, c-Crk show less
Epitope
AA 102-304
(25), (17), (6), (5), (2), (1), (1), (1), (1), (1), (1), (1)
Reactivity
Human
(109), (59), (58), (37), (24), (24), (12), (2), (2), (1)
Host
Mouse
(87), (22)
Clonality (Clone)
Monoclonal ()
Conjugate
Un-conjugated
(4), (4), (4), (3), (3), (3), (3), (3), (3), (3), (3), (1), (1), (1)
Application
Western Blotting (WB), Immunoprecipitation (IP), BioImaging (BI)
(64), (35), (30), (24), (19), (18), (15), (13), (6), (3), (2), (2), (1)
Pubmed 7 references available
Quantity 50 µg
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Catalog No. ABIN967702
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Immunogen Human Crk
Clone LPR-02
Isotype IgG2a
Cross-Reactivity Cow (Bovine), Chicken, Dog (Canine), Frog, Mouse (Murine), Rat (Rattus)
Characteristics 1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
2. Please refer to us for technical protocols.
3. This antibody has been developed and certified for the bioimaging application. However, a routine bioimaging test is not performed on every lot. Researchers are encouraged to titrate the reagent for optimal performance.
4. Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
5. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
6. Triton is a trademark of the Dow Chemical Company.
Purification Purified from tissue culture supernatant or ascites by affinity chromatography.
Alternative Name Crk
Background Crk was first isolated as the v-crk oncogene from chicken retroviruses CT10 and ASV-1. All human cell lines examined to date express a 40 kDa Crk protein. In addition, there is variable expression of 42 kDa and 28 kDa Crk proteins. The c-crk gene is one of a class of genes, such as Nck and GRB2/ASH, which encode proteins that consist mainly of SH2 and SH3 domains. These proteins function as adaptor molecules in tyrosine kinase signal transduction pathways. The SH2 domains interact with phosphotyrosine-containing peptides, while the SH3 domains can enhance this interaction and/or bind to other cellular components. Both the SH2 and SH3 domains of the human Crk protein are required for differentiation of PC12 cells. Thus, Crk has a role in an NGF-induced signaling pathway that involves activation of p21ras. Furthermore, three proteins of 118 kDa, 125 kDa, and 136 kDa which specifically bind to the Crk SH3 domain have been identified.
Molecular Weight 40 kDa
Application Notes Bioimaging
1. Seed the cells in appropriate culture medium at ~10,000 cells per well in an 96-well Imaging Plate and culture overnight.
2. Remove the culture medium from the wells, and fix the cells by adding 100 myl of Fixation Buffer to each well. Incubate for 10 minutes at room temperature (RT). 3. Remove the fixative from the wells, and permeabilize the cells using either 90% methanol, or Triton™ X-100: a. Add 100 myl of -20°C 90% methanol or to each well and incubate for 5 minutes at RT. OR b. Add 100 myl of 0.1% Triton™ X-100 to each well and incubate for 5 minutes at RT.
4. Remove the permeabilization buffer, and wash the wells twice with 100 myl of 1× PBS.
5. Remove the PBS, and block the cells by adding 100 myl of to each well. Incubate for 30 minutes at RT.
6. Remove the blocking buffer and add 50 myl of the optimally titrated primary antibody (diluted in Stain Buffer) to each well, and incubate for 1 hour at RT.
7. Remove the primary antibody, and wash the wells three times with 100 myl of 1× PBS.
8. Remove the PBS, and add the second step reagent at its optimally titrated concentration in 50 myl to each well, and incubate in the dark for 1 hour at RT.
9. Remove the second step reagent, and wash the wells three times with 100 myl of 1× PBS.
10. Remove the PBS, and counter-stain the nuclei by adding 200 myl per well of 2 myg/ml Hoechst 33342 in 1× PBS to each well at least 15 minutes before imaging.
11. View and analyze the cells on an appropriate imaging instrument.
Comment

Related Products: ABIN967389, ABIN968535

Restrictions For Research Use only
Format Liquid
Concentration 250 µg/ml
Buffer Aqueous buffered solution containing BSA, glycerol.
Preservative Sodium azide
Precaution of Use This product contains sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
Storage -20 °C
Supplier Images
anti-V-Crk Sarcoma Virus CT10 Oncogene Homolog (Avian) (CRK) (AA 102-304) antibody Western blot analysis of Crk on a HeLa lysate. Lane 1: 1:5000, lane 2: 1:10000, lane 1:20000 dilution of the Crk antibody.
anti-V-Crk Sarcoma Virus CT10 Oncogene Homolog (Avian) (CRK) (AA 102-304) antibody (2) Immunofluorescent staining of HeLa (ATCC CCL-2) cells. Cells were seeded in a 96 well imaging plate at ~ 10 000 cells per well. After overnight incubation, cells were stained using the Triton™ X-100 perm protocol and the anti-Crk antibody. The second step reagent was FITC goat anti mouse Ig. Images were taken on a BD Pathway 855 Bioimager system using a 20x objective. This antibody also stained A549 (ATCC CCL-185) and U-2 OS (ATCC HTB-96) cells and worked with both the Triton™ X-100 and alcohol perm protocols.
Product cited in: Fournier, Lamorte, Maroun et al.: "Cbl-transforming variants trigger a cascade of molecular alterations that lead to epithelial mesenchymal conversion." in: Molecular biology of the cell, Vol. 11, Issue 10, pp. 3397-410, 2000 (PubMed).

Smith, Evans, Murakami et al.: "Wee1-regulated apoptosis mediated by the crk adaptor protein in Xenopus egg extracts." in: The Journal of cell biology, Vol. 151, Issue 7, pp. 1391-400, 2001 (PubMed).

Girardin, Yaniv: "A direct interaction between JNK1 and CrkII is critical for Rac1-induced JNK activation." in: The EMBO journal, Vol. 20, Issue 13, pp. 3437-46, 2001 (PubMed).

Cho, Klemke: "Purification of pseudopodia from polarized cells reveals redistribution and activation of Rac through assembly of a CAS/Crk scaffold." in: The Journal of cell biology, Vol. 156, Issue 4, pp. 725-36, 2002 (PubMed).

Smith, Richardson, Kopf et al.: "Apoptotic regulation by the Crk adapter protein mediated by interactions with Wee1 and Crm1/exportin." in: Molecular and cellular biology, Vol. 22, Issue 5, pp. 1412-23, 2002 (PubMed).

Liu, Kimura, Baumann et al.: "APS facilitates c-Cbl tyrosine phosphorylation and GLUT4 translocation in response to insulin in 3T3-L1 adipocytes." in: Molecular and cellular biology, Vol. 22, Issue 11, pp. 3599-609, 2002 (PubMed).

Miller, Chen, Gharib et al.: "Increased C-CRK proto-oncogene expression is associated with an aggressive phenotype in lung adenocarcinomas." in: Oncogene, Vol. 22, Issue 39, pp. 7950-7, 2003 (PubMed).

Validation Images
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