![Full length untagged recombinant human Lck protein was either not added (-) or added (+) to control cell extract which does not contain Lck and then resolved by SDS-PAGE on a 10% Tris-glycine gel. The proteins then were transferred to nitrocellulose and incubated with 0.50 μg/mL Anti-Lck [38-54] pan antibody. After washing, membranes were incubated with goat F(ab')2 anti-rabbit IgG alkaline phosphatase and bands were detected using the Tropix WesternStarTM detection method. The data demonstrate the specificity of the antibody for the Lck protein.](/media/images/0094731.jpeg)
BP7117 Lck antibody
see related secondary antibodiessee all 59 Lck products
0.1 mg / US$ 515
NOVUS BIOLOGICALS
PO Box 802 Littleton, CO 80160, USA
E-Mail: novus@novus-biologicals.com
Homepage: http://www.novus-biologicals.com
PO Box 802 Littleton, CO 80160, USA
E-Mail: novus@novus-biologicals.com
Homepage: http://www.novus-biologicals.com
Quick Overview
Rabbit anti Lck
Synonyms
p56,
Product Description
Rabbit anti Lck , Presentation: Aff - Purified. Product is tested for Western blot / Immunoblot ( WB )
Properties
| Applications | Western blot / Immunoblot ( WB ) |
| Reactivity | Mouse ( Ms ), Human ( Hu ) |
| Presentation | Aff - Purified |
| Host | Rabbit |
| Catalog Number | BP7117 |
| Swiss Prot. No. | P06239 |
| Quantity | 0.1 mg |
| Price | US$ 515 |
| Delivery | Worldwide |
| Manufacturer | Acris Antibodies GmbH |
| Datasheet | view  BP7117.pdf |
Datasheet Extract
| Background | Lck (p56lck), a member of the Src family of non-receptor tyrosine protein kinases, is expressed predominantly in T cells. Lck function is critical both for T cell development in the thymus and activation of mature T cells in the periphery by antigen. The activity of Lck is known to be regulated by phosphorylation of two conserved tyrosine residues, Tyr-505 (equivalent to Tyr-529 in c-Src) and Tyr-394 (equivalent to Tyr-418 in c-Src). |
| Immunogen | Chemically synthesized peptide derived from amino acid region 38 to 54 of Lck protein Swiss Num.: P06239 Remarks: The sequence is conserved in human and mouse. |
| Format | State: Liquid Ig fraction Purification: Epitope-specific chromatography BufferSystem: Phosphate buffer, pH 7.4, with 1.0 mg/mL BSA (IgG, protease-free) as a carrier containing 0.05 % sodium azide as preservative |
| Applications | Western blot (0.1-1.0 μg/mL; at 0.50 μg/mL, the dilution provides 200 mL working solution, which at 10 mL/blot allows 20 blots to be performed). |
| Specificity | Anti-Lck [38-54] Pan antibody is useful to determine total levels of Lck protein. Species: Human, mouse. |
| Storage | Store at -80 ºC. Upon initial thawing, aliquot and store at -80 ºC. Avoid repeated freezing and thawing.
Shelf life: one year from despatch. |
| References | Fujimaki, W., et al. (2001) Functional uncoupling of TCR engagement and Lck activation in anergic human thymic CD4{super+} T cells. J. Biol. Chem. 276(20):17455-17460.
Ellery, J.M., et al. (2000) Activation of the interleukin 2 receptor: a possible role for tyrosine phosphatases. Cell Signal 12(6):367-373. Gervais, F.G. and A. Veillette (1997) Reconstitution of interactions between protein-tyrosine phosphatase CD45 and tyrosine-protein kinase p56(lck) in nonlymphoid cells. J. Biol. Chem. 272(19):12754-12761. Yamaguchi, H. and W.A. Hendrickson (1996) Structural basis for activation of human lymphocyte kinase Lck upon tyrosine phosphorylation. Nature 384(6608):484-489. |
| Protocols | Western Blotting Procedure
1. Lyse approximately 10e7 cells in 0.5 mL of ice cold Cell Lysis Buffer (formulation provided below). This buffer, a modified RIPA buffer, is suitable for recovery of most proteins, including membrane receptors, cytoskeletal-associated proteins, and soluble proteins. Other cell lysis buffer formulations, such as Laemmli sample buffer and Triton-X 100 buffer, are also compatible with this procedure. Additional optimization of the cell stimulation protocol and cell lysis procedure may be required for each specific application. 2. Remove the cellular debris by centrifuging the lysates at 14,000 x g for 10 minutes. Alternatively, lysates may be ultracentrifugedat 100,000 x g for 30 minutes for greater clarification. 3. Carefully decant the clarified cell lysates into clean tubes and determine the protein concentration using a suitable method, such as the Bradford assay. Polypropylene tubes are recommended for storing cell lysates. 4. React an aliquot of the lysate with an equal volume of 2x Laemmli Sample Buffer (125 mM Tris, pH 6.8, 10% glycerol, 10% SDS, 0.006% bromophenol blue, and 130 mM dithiothreitol [DTT]) and boil the mixture for 90 seconds at 100°C. 5. Load 10-30 µg of the cell lysate into the wells of an appropriate single percentage or gradient minigel and resolve the proteins by SDS-PAGE. 6. In preparation for the Western transfer, cut a piece of PVDF membrane slightly larger than the gel. Soak the membrane in methanol for 1 minute, then rinse with ddH2O for 5 minutes. Alternatively, nitrocellulose may be used. 7. Soak the membrane, 2 pieces of Whatman paper, and Western apparatus sponges in transfer buffer (formulation provided below) for 2 minutes. 8. Assemble the gel and membrane into the sandwich apparatus. 9. Transfer the proteins at 140 mA for 60-90 minutes at room temperature. 10. Following the transfer, rinse the membrane with Tris buffered saline for 2 minutes. 11. Block the membrane with blocking buffer (formulation provided below) for one hour at room temperature or overnight at 4°C. 12. Incubate the blocked blot with primary antibody at a concentration of 0.1-1.0 μg/mL in Tris buffered saline supplemented with 3% BSA and 0.1% Tween 20 for 2 hours at room temperature. 13. Wash the blot with several changes of Tris buffered saline supplemented with 0.1% Tween 20. 14. Detect the antibody band using an appropriate secondary antibody, such as goat F(ab)2 anti-rabbit IgG alkaline phosphatase conjugate or goat F(ab)2 anti-rabbit IgG horseradish peroxidase conjugate in conjunction with your chemiluminescence reagents and instrumentation. Cell Lysis Buffer Formulation: 10 mM Tris, pH 7.4 100 mM NaCl 1 mM EDTA 1 mM EGTA 1 mM NaF 20 mM Na4P2O7 2 mM Na3VO4 0.1% SDS 0.5% sodium deoxycholate 1% Triton-X 100 10% glycerol 1 mM PMSF (made from a 0.3 M stock in DMSO) or 1 mM AEBSF (water soluble version of PMSF) 60 µg/mL aprotinin 10 µg/mL leupeptin 1 µg/mL pepstatin (alternatively, protease inhibitor cocktail such as Sigma Cat. # P2714 may be used) Transfer Buffer Formulation: 2.4 gm Tris base 14.2 gm glycine 200 mL methanol Q.S. to 1 liter, then add 1 mL 10% SDS. Cool to 4°C prior to use. Tris Buffered Saline Formulation: 20 mM Tris-HCl, pH 7.4 0.9% NaCl Blocking Buffer Formulation: 100 mL Tris buffered saline 5 gm BSA 0.1 mL Tween 20 |
| Pictures | Full length untagged recombinant human Lck protein was either not added (-) or added (+) to control cell extract which does not contain Lck and then resolved by SDS-PAGE on a 10% Tris-glycine gel. The proteins then were transferred to nitrocellulose and incubated with 0.50 μg/mL Anti-Lck [38-54] pan antibody. After washing, membranes were incubated with goat F(ab')2 anti-rabbit IgG alkaline phosphatase and bands were detected using the Tropix WesternStarTM detection method. The data demonstrate the specificity of the antibody for the Lck protein. |
12 Secondary Antibodies
| Catalog No. | Name / Host | Presentation | Reactivity | ||||
|---|---|---|---|---|---|---|---|
| R1364B | Rabbit IgG (H&L) | ||||||
| Goat | Biotin | 2 mg / US$ 295 | |||||
| R1364F | Rabbit IgG (H&L) | ||||||
| Goat | FITC | 2 mg / US$ 285 | |||||
| R1364T | Rabbit IgG (H&L) | ||||||
| Goat | TRITC | 2 mg / US$ 285 | |||||
| R1364TR | Rabbit IgG (H&L) | ||||||
| Goat | Texas Red | 2 mg / US$ 295 | |||||
| R1364HRP | Rabbit IgG (H&L) | ||||||
| Goat | HRP | 2 mg / US$ 295 | |||||
| R1364AP | Rabbit IgG (H&L) | ||||||
| Goat | AP | 1 mg / US$ 325 | |||||
| R1458C2 | Rabbit IgG (H&L) multi-species ads. | ||||||
| Goat | Cy2 | 1 mg / US$ 445 | |||||
| R1458C3 | Rabbit IgG (H&L) multi-species ads. | ||||||
| Goat | Cy3 | 1 mg / US$ 445 | |||||
| R1458C35 | Rabbit IgG (H&L) multi-species ads. | ||||||
| Goat | Cy3.5 | 1 mg / US$ 445 | |||||
| R1458C5 | Rabbit IgG (H&L) multi-species ads. | ||||||
| Goat | Cy5 | 1 mg / US$ 445 | |||||
| R1458C55 | Rabbit IgG (H&L) multi-species ads. | ||||||
| Goat | Cy5.5 | 1 mg / US$ 445 | |||||
| R1427R | Rabbit IgG (H&L) F(ab')2 Fragment multi-species ads. | ||||||
| Donkey | PE | 1 ml / US$ 455 | |||||
Click here to see all secondary antibodies for 'BP7117 Lck'.
