Phospho-LYN (Y397)/LCK (Y394)/HCK (Y411)/BLK (Y389) Recombinant Rabbit Monoclonal Antibody [PSH09-43]
cat.: HA723105
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat
Applications: WB, IF-Cell
Clonality: Monoclonal
Clone number: PSH09-43
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
Storage buffer: PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration: 1ug/ul
Purification: Protein A affinity purified.
Molecular weight: Predicted band size: 58 kDa
Isotype: IgG
Immunogen: Synthetic phospho-peptide corresponding to residues surrounding Tyr394 of human LCK.
Positive control: COLO205 cell lysate, TT cell lysate, Ramos serum starved for 16 hours cell lysate, Ramos serum starved for 16 hours add 12μg/mL human IgM (diluted in serum free medium) for 2 minutes cell lysate, NIH/3T3 cell lysate, NIH/3T3 treated with 50mM sodium orthovanadate for 5 minutes cell lysate, PC-12 cell lysate, PC-12 treated with 1mM sodium orthovanadate for 30 minutes cell lysate, Ramos cells serum starved for 16 hours add 12μg/mL human IgM (diluted in serum free medium) for 2 minutes.
Subcellular location: Cell membrane, Cytoplasm, cytosol.
Recommended Dilutions:
  WB
  IF-Cell
1:2,000
1:100
Uniprot #: SwissProt: P07948 Human | P06239 Human | P08631 Human | P51451 Human
Alternative names: AA407514 EC 2.7.10.2 FLJ26625 Hck 2 JTK 8 JTK8 Lck/Yes related novel protein tyrosine kinase LYN LYN proto oncogene, Src family tyrosine kinase LYN_HUMAN ONCOGENE LYN p53Lyn p56Lyn Tyrosine protein kinase LYN Tyrosine-protein kinase Lyn V yes 1 Yamaguchi sarcoma viral related oncogene homolog Yamaguchi sarcoma viral (v yes 1) related oncogene homolog IMD22 LCK Lck p56 LCK proto-oncogene, Src family tyrosine kinase LCK_HUMAN Leukocyte C-terminal Src kinase LSK Lymphocyte cell specific protein tyrosine kinase Lymphocyte cell-specific protein-tyrosine kinase Lymphocyte specific protein tyrosine kinase Membrane associated protein tyrosine kinase Oncogene lck P56 LCK p56(LSTRA) protein tyrosine kinase p56-LCK p56lck pp58 lck pp58lck Protein YT16 Proto oncogene tyrosine protein kinase LCK Proto-oncogene Lck Protooncogene tyrosine protein kinase LCK T cell specific protein tyrosine kinase T cell-specific protein-t......
Images
HA723105_1.jpg Fig1: Western blot analysis of Phospho-LYN (Y397)/LCK (Y394)/HCK (Y411)/BLK (Y389) on different lysates with Rabbit anti-Phospho-LYN (Y397)/LCK (Y394)/HCK (Y411)/BLK (Y389) antibody (HA723105) at 1/2,000 dilution.

Lane 1: COLO205 cell lysate
Lane 2: TT cell lysate (negative)
Lane 3: Ramos serum starved for 16 hours cell lysate
Lane 4: Ramos serum starved for 16 hours add 12μg/mL human IgM (diluted in serum free medium) for 2 minutes cell lysate
Lane 5: Ramos serum starved for 16 hours add 12μg/mL human IgM (diluted in serum free medium) for 2 minutes cell lysate, then the membrane treated with λpp for 1 hour

Lysates/proteins at 20 µg/Lane.

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

Exposure time: 23 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 (HA723105) 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.
HA723105_2.jpg Fig2: Western blot analysis of Phospho-LYN (Y397)/LCK (Y394)/HCK (Y411)/BLK (Y389) on different lysates with Rabbit anti-Phospho-LYN (Y397)/LCK (Y394)/HCK (Y411)/BLK (Y389) antibody (HA723105) at 1/2,000 dilution.

Lane 1: NIH/3T3 cell lysate (20 µg/Lane)
Lane 2: NIH/3T3 treated with 50mM sodium orthovanadate for 5 minutes cell lysate (20 µg/Lane)
Lane 3: PC-12 cell lysate (20 µg/Lane)
Lane 4: PC-12 treated with 1mM sodium orthovanadate for 30 minutes cell lysate (20 µg/Lane)

Predicted band size: 58 kDa
Observed band size: 58 kDa
Exposure time: Lane 1-2: 20 seconds; Lane 3-4: 1 minute; ECL: K1802;
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 (HA723105) 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.
HA723105_3.jpg Fig3: Immunocytochemistry analysis of Ramos cells serum starved for 16 hours add 12μg/mL human IgM (diluted in serum free medium) for 2 minutes labeling Phospho-LYN (Y397)/LCK (Y394)/HCK (Y411)/BLK (Y389) with Rabbit anti-Phospho-LYN (Y397)/LCK (Y394)/HCK (Y411)/BLK (Y389) antibody (HA723105) at 1/100 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-Phospho-LYN (Y397)/LCK (Y394)/HCK (Y411)/BLK (Y389) antibody (HA723105) at 1/100 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.
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.