Lck Recombinant Rabbit Monoclonal Antibody [PSH09-25]
Catalog# HA723083
Lck Recombinant Rabbit Monoclonal Antibody [PSH09-25]
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WB
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IF-Cell
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IHC-P
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IP
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FC
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Human
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Mouse
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Rat
概述
产品名称
Lck Recombinant Rabbit Monoclonal Antibody [PSH09-25]
抗体类型
Recombinant Rabbit monoclonal Antibody
免疫原
Recombinant protein within human Lck aa 1-150.
种属反应性
Human, Mouse, Rat
验证应用
WB, IF-Cell, IHC-P, IP, FC
分子量
Predicted band size: 58 kDa
阳性对照
Ramos cell lysate, Raji cell lysate, Jurkat cell lysate, MOLT-4 cell lysate, Mouse spleen tissue lysate, Rat spleen tissue lysate, Jurkat, mouse spleen tissue, rat spleen tissue.
偶联
unconjugated
克隆号
PSH09-25
产品特性
形态
Liquid
存放说明
Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term.
存储缓冲液
PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
亚型
IgG
纯化方式
Protein A affinity purified.
应用稀释度
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WB
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1:2,000
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IF-Cell
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1:500
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IHC-P
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1:1,000
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IP
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1-2μg/sample
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FC
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1:1,000
靶点
功能
Tyrosin-protein kinase Lck (or lymphocyte-specific protein tyrosine kinase) is a 56 kDa protein that is found inside lymphocytes and encoded in the human by the LCK gene. The Lck is a member of Src kinase family (SFK) and is important for the activation of T-cell receptor (TCR) signaling in both naive T cells and effector T cells. The role of Lck is less prominent in the activation or in the maintenance of memory CD8 T cells in comparison to CD4 T cells. In addition, the constitutive activity of the mouse Lck homolog varies among memory T cell subsets. It seems that in mice, in the effector memory T cell (TEM) population, more than 50% of Lck is present in a constitutively active conformation, whereas less than 20% of Lck is present as active form in central memory T cells. These differences are due to differential regulation by SH2 domain–containing phosphatase-1 (Shp-1) and C-terminal Src kinase. Lck is responsible for the initiation of the TCR signaling cascade inside the cell by phosphorylating immunoreceptor tyrosine-based activation motifs (ITAM) within the TCR-associated chains. Lck can be found in different forms in immune cells: free in the cytosol or bound to the plasma membrane (PM) through myristoylation and palmitoylation. Due to the presence of the conserved CxxC motif (C20 and C23) in the zinc clasp structure, Lck is able to bind the cell surface coreceptors CD8 and\or CD4. Bound and free Lck have different properties: free Lck has more pronounced kinase activity in comparison to bound Lck, and moreover, the free form produces a higher level of T cell activation. The reasons for these differences are not well understood yet.
背景文献
1. Wu J et al. Asparagine enhances LCK signalling to potentiate CD8(+) T-cell activation and anti-tumour responses. Nat Cell Biol. 2021 Jan
2. Wu L et al. CD28-CAR-T cell activation through FYN kinase signaling rather than LCK enhances therapeutic performance. Cell Rep Med. 2023 Feb
亚细胞定位
Cell membrane, Cytoplasm, cytosol.
别名
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
展开图片
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☑ Relative expression (RE)
Western blot analysis of Lck on different lysates with Rabbit anti-Lck antibody (HA723083) at 1/2,000 dilution.
Lane 1: Ramos cell lysate (20 µg/Lane)
Lane 2: Raji cell lysate (20 µg/Lane)
Lane 3: HeLa cell lysate (negative) (20 µg/Lane)
Lane 4: Jurkat cell lysate (20 µg/Lane)
Lane 5: MOLT-4 cell lysate (20 µg/Lane)
Lane 6: Mouse spleen tissue lysate (40 µg/Lane)
Lane 7: Rat spleen tissue lysate (40 µg/Lane)
Predicted band size: 58 kDa
Observed band size: 55 kDa
Exposure time: 30 seconds; ECL: K1801;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA723083) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. -
☑ Relative expression (RE)
Immunocytochemistry analysis of Jurkat (positive) and HeLa (negative) labeling Lck with Rabbit anti-Lck antibody (HA723083) at 1/500 dilution.
Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Lck antibody (HA723083) at 1/500 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.
Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution. -
Immunohistochemical analysis of paraffin-embedded mouse spleen tissue with Rabbit anti-Lck antibody (HA723083) at 1/1,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA723083) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Immunohistochemical analysis of paraffin-embedded rat spleen tissue with Rabbit anti-Lck antibody (HA723083) at 1/1,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA723083) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Lck was immunoprecipitated from 0.2 mg Jurkat cell lysate with HA723083 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA723083 at 1/1,000 dilution. Mouse Anti-Rabbit IgG kappa light chain secondary antibody (M1208-2) at 1/5,000 dilution was used for 1 hour at room temperature.
Lane 1: Jurkat cell lysate (input)
Lane 2: HA723083 IP in Jurkat cell lysate
Lane 3: Rabbit IgG instead of HA723083 in Jurkat cell lysate
Blocking/Dilution buffer: 5% NFDM/TBST
Exposure time: 4 seconds; ECL: K1801 -
☑ Relative expression (RE)
Flow cytometric analysis of HeLa (left, negative) and Jurkat (right, positive) cells labeling Lck.
Cells were fixed and permeabilized. Then stained with the primary antibody (HA723083, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
请注意: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"
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