Recombinant Anti-Lck antibody [EPR20798-107] KO Tested (ab227975)

Product name

Anti-Lck antibody [EPR20798-107]
See all Lck primary antibodies
Description

Rabbit monoclonal [EPR20798-107] to Lck
Host species

Rabbit
Tested applications

Suitable for: WB, IP, IHC-P, ICC/IF, Flow Cyt (Intra)more details
Species reactivity

Reacts with: Mouse, Rat, Human
Immunogen

Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: WEHI-231, Raji, Ramos and Jurkat whole cell lysates; Human thymus and tonsil lysates; Rat thymus lysate; Mouse lymph node lysate. IHC-P: Human diffuse large B-cell lymphoma tissue; Mouse spleen tissue; Rat colon tissue. ICC/IF: Ramos and WEHI-231 cells. Flow Cyt (intra): Jurkat and WEHI-231 cells. IP: Jurkat whole cell lysate.
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
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

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: 7.2
Preservative: 0.01% Sodium azide
Constituents: 0.05% BSA, 40% Glycerol (glycerin, glycerine), PBS
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR20798-107
Isotype

IgG
Research areas
- Neuroscience
- Development
- Neuroscience
- Processes

Alternative Versions
- Anti-Lck antibody [EPR20798-107] - BSA and Azide free (ab229379)
Compatible Secondaries
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
Recombinant Protein
- Recombinant human Lck protein (ab79626)
Related Products
- Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (ab195889)
- Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) (ab93684)

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab227975 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
WB		1/1000. Detects a band of approximately 58 kDa (predicted molecular weight: 58 kDa).
IP		1/30.
IHC-P		1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF		1/100.
Flow Cyt (Intra)		1/500.

Notes
WB 1/1000. Detects a band of approximately 58 kDa (predicted molecular weight: 58 kDa).
IP 1/30.
IHC-P 1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF 1/100.
Flow Cyt (Intra) 1/500.

Function

Tyrosine kinase that plays an essential role for the selection and maturation of developing T-cell in the thymus and in mature T-cell function. Is constitutively associated with the cytoplasmic portions of the CD4 and CD8 surface receptors and plays a key role in T-cell antigen receptor(TCR)-linked signal transduction pathways. Association of the TCR with a peptide antigen-bound MHC complex facilitates the interaction of CD4 and CD8 with MHC class II and class I molecules, respectively, and thereby recruits the associated LCK to the vicinity of the TCR/CD3 complex. LCK then phosphorylates tyrosines residues within the immunoreceptor tyrosines-based activation motifs (ITAMs) in the cytoplasmic tails of the TCRgamma chains and CD3 subunits, initiating the TCR/CD3 signaling pathway. In addition, contributes to signaling by other receptor molecules. Associates directly with the cytoplasmic tail of CD2, and upon engagement of the CD2 molecule, LCK undergoes hyperphosphorylation and activation. Also plays a role in the IL2 receptor-linked signaling pathway that controls T-cell proliferative response. Binding of IL2 to its receptor results in increased activity of LCK. Is expressed at all stages of thymocyte development and is required for the regulation of maturation events that are governed by both pre-TCR and mature alpha beta TCR. Phosphorylates RUNX3.
Tissue specificity

Expressed specifically in lymphoid cells.
Involvement in disease

Note=A chromosomal aberration involving LCK is found in leukemias. Translocation t(1;7)(p34;q34) with TCRB.
Sequence similarities

Belongs to the protein kinase superfamily. Tyr protein kinase family. SRC subfamily.
Contains 1 protein kinase domain.
Contains 1 SH2 domain.
Contains 1 SH3 domain.
Domain

The SH2 domain mediates interaction with SQSTM1. Interaction is regulated by Ser-59 phosphorylation.
Post-translational
modifications

Phosphorylated on Tyr-394, which increases enzymatic activity (By similarity). Phosphorylated on Tyr-505, which decreases activity.
Cellular localization

Cytoplasm. Cell membrane. Present in lipid rafts in an unactive form.
Target information above from: UniProt accession P06239 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
Database links
- Entrez Gene: 3932 Human
- Entrez Gene: 16818 Mouse
- Entrez Gene: 313050 Rat
- Omim: 153390 Human
- SwissProt: P06239 Human
- SwissProt: P06240 Mouse
- SwissProt: Q01621 Rat
- Unigene: 470627 Human
Form

This protein is known to be similar in amino acid sequence to HCK (P08631), FYN (P06241), YES1 (P07947), SRC (P12931), and LYN (P07948). Therefore, cross-reactivity with these homologous proteins may be observed. We would be happy to provide immunogen alignment information upon request.
Alternative names
- IMD22 antibody
- LCK antibody
- Lck p56 antibody
- LCK proto-oncogene, Src family tyrosine kinase antibody
- LCK_HUMAN antibody
- Leukocyte C-terminal Src kinase antibody
- LSK antibody
- Lymphocyte cell specific protein tyrosine kinase antibody
- Lymphocyte cell-specific protein-tyrosine kinase antibody
- Lymphocyte specific protein tyrosine kinase antibody
- Membrane associated protein tyrosine kinase antibody
- Oncogene lck antibody
- P56 LCK antibody
- p56(LSTRA) protein tyrosine kinase antibody
- p56-LCK antibody
- p56lck antibody
- pp58 lck antibody
- pp58lck antibody
- Protein YT16 antibody
- Proto oncogene tyrosine protein kinase LCK antibody
- Proto-oncogene Lck antibody
- Protooncogene tyrosine protein kinase LCK antibody
- T cell specific protein tyrosine kinase antibody
- T cell-specific protein-tyrosine kinase antibody
- T lymphocyte specific protein tyrosine kinase p56lck antibody
- Tyrosine-protein kinase Lck antibody
- YT 16 antibody
- YT16 antibody
see all

Western blot - Anti-Lck antibody [EPR20798-107] (ab227975)

All lanes : Anti-Lck antibody [EPR20798-107] (ab227975) at 1/1000 dilution

Lane 1 : Wild-type Jurkat cell lysate
Lane 2 : Lck knockout Jurkat cell lysate
Lane 3 : Ramos cell lysate
Lane 4 : A549 cell lysate

Lysates/proteins at 20 µg per lane.

Performed under reducing conditions.

Predicted band size: 58 kDa
Observed band size: 60 kDa why is the actual band size different from the predicted?

False colour image of Western blot: Anti-Lck antibody [EPR20798-107] staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab227975 was shown to bind specifically to Lck. A band was observed at 60 kDa in wild-type Jurkat cell lysates with no signal observed at this size in Lck knockout cell line ab273855 (knockout cell lysate ab273809). To generate this image, wild-type and Lck knockout Jurkat 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 (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.
Western blot - Anti-Lck antibody [EPR20798-107] (ab227975)

All lanes : Anti-Lck antibody [EPR20798-107] (ab227975) at 1/1000 dilution

Lane 1 : Human thymus tissue lysate
Lane 2 : Human tonsil tissue lysate
Lane 3 : WEHI-231 (mouse lymphoblast B cell lymphoma cell line) whole cell lysate
Lane 4 : Rat thymus tissue lysate
Lane 5 : Mouse lymph node tissue lysate
Lane 6 : Raji (human Burkitt's lymphoma cell line) whole cell lysate
Lane 7 : Ramos (human Burkitt's lymphoma cell line) whole cell lysate
Lane 8 : Jurkat (human T cell leukemia cell line from peripheral blood) 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

Developed using the ECL technique.

Predicted band size: 58 kDa
Observed band size: 58 kDa

Exposure times: Lanes 1,2,7: 5 seconds; Lanes 3,4,8: 1 second; Lane 5: 3 seconds; Lane 6: 10 seconds.

Blocking/Dilution buffer: 5% NFDM/TBST.
Immunoprecipitation - Anti-Lck antibody [EPR20798-107] (ab227975)

Lck was immunoprecipitated from 0.35 mg of Jurkat (human T cell leukemia cell line from peripheral blood) whole cell lysate with ab227975 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab227975 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

Lane 1: Jurkat whole cell lysate 10 µg (Input).
Lane 2: ab227975 IP in Jurkat whole cell lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab227975 in Jurkat whole cell lysate.

Exposure time: 8 seconds.

Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Flow Cytometry (Intracellular) - Anti-Lck antibody [EPR20798-107] (ab227975)

Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol-permeabilized WEHI-231 (mouse lymphoblast B cell lymphoma cell line) cell line labeling Lck with ab227975 at 1/500 dilution (red) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) at 1/2000 dilution was used as the secondary antibody.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Lck antibody [EPR20798-107] (ab227975)

Immunohistochemical analysis of paraffin-embedded rat colon tissue labeling Lck with ab227975 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP), ready to use. Membranous and cytoplasmic staining in T cells of rat colon is observed (PMID: 16769579). Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP), ready to use.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Flow Cytometry (Intracellular) - Anti-Lck antibody [EPR20798-107] (ab227975)

Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol permeabilized Jurkat (human T cell leukemia cell line from peripheral blood) cell line labeling Lck with ab227975 at 1/500 dilution (red) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) at 1/2000 dilution was used as the secondary antibody.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Lck antibody [EPR20798-107] (ab227975)

Immunohistochemical analysis of paraffin-embedded mouse spleen tissue labeling Lck with ab227975 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP), ready to use. Membranous and cytoplasmic staining in mouse spleen reactive lymph node and T cells is observed (PMID: 16769579). Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP), ready to use.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Lck antibody [EPR20798-107] (ab227975)

Immunohistochemical analysis of paraffin-embedded human diffuse large B-cell lymphoma tissue labeling Lck with ab227975 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP), ready to use. Membranous and cytoplasmic staining in human diffuse large B-cell lymphoma is observed (PMID:16769579). Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP), ready to use.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunocytochemistry/ Immunofluorescence - Anti-Lck antibody [EPR20798-107] (ab227975)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized WEHI-231 (mouse lymphoblast B cell lymphoma cell line) cells labeling Lck with ab227975 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing membranous staining on WEHI-231 cell line.

The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) at 1/200 dilution (red).

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) at 1/1000 dilution.
Immunocytochemistry/ Immunofluorescence - Anti-Lck antibody [EPR20798-107] (ab227975)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized Ramos (human Burkitt's lymphoma cell line) cells labeling Lck with ab227975 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing membranous staining on Ramos cell line.

The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) at 1/200 dilution (red).

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) at 1/1000 dilution.
Anti-Lck antibody [EPR20798-107] (ab227975)

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.

Click here to view the general protocols

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Key features and details

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Overview

Product name

Description

Host species

Tested applications

Species reactivity

Immunogen

Positive control

General notes

Properties

Form

Storage instructions

Storage buffer

Purity

Clonality

Clone number

Isotype

Research areas

Associated products

Alternative Versions

Compatible Secondaries

Isotype control

Recombinant Protein

Related Products

Applications

The Abpromise guarantee

Target

Function

Tissue specificity

Involvement in disease

Sequence similarities

Domain

Post-translationalmodifications

Cellular localization

Database links

Form

Alternative names

Images

Protocols

Datasheets and documents

SDS download

Datasheet download

Certificate of Compliance

References (0)

Customer reviews and Q&As

Post-translational
modifications