PRODUCT CODE: ET1702-03

Src Recombinant Rabbit Monoclonal Antibody [JF0947] (ET1702-03)

  • Recombinant

Applications

  • WB

  • ICC

  • IF

  • IHC-P

  • FC

REACTIVITY

  • Human

  • Mouse

  • Rat

-
+
Western blot analysis of Src on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1702-03, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.<br />
Positive control: <br />
Lane 1: HUVEC cell lysate<br />
Lane 2: A549 cell lysate<br />
Lane 3: PC-3M cell lysate<br />
Lane 4: SH-SY5Y cell lysate
  • Western blot analysis of Src on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1702-03, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.<br />
Positive control: <br />
Lane 1: HUVEC cell lysate<br />
Lane 2: A549 cell lysate<br />
Lane 3: PC-3M cell lysate<br />
Lane 4: SH-SY5Y cell lysate
  • ICC staining of Src in Hela cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1702-03, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
  • ICC staining of Src in MCF-7 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1702-03, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
  • ICC staining of Src in A431 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1702-03, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
  • Immunohistochemical analysis of paraffin-embedded human tonsil tissue using anti-Src antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1702-03, 1/50) for 30 minutes 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 human breast carcinoma tissue using anti-Src antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1702-03, 1/50) for 30 minutes 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 human kidney tissue using anti-Src antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1702-03, 1/50) for 30 minutes 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 brain tissue using anti-Src antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1702-03, 1/50) for 30 minutes 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 mouse kidney tissue using anti-Src antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1702-03, 1/50) for 30 minutes 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.
  • Flow cytometric analysis of Src was done on NIH/3T3 cells. The cells were fixed, permeabilized and stained with the primary antibody (ET1702-03, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Rabbit IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
Western blot analysis of Src on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1702-03, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.
Positive control:
Lane 1: HUVEC cell lysate
Lane 2: A549 cell lysate
Lane 3: PC-3M cell lysate
Lane 4: SH-SY5Y cell lysate

Applications

  • WB

  • ICC

  • IF

  • IHC-P

  • FC

REACTIVITY

  • Human

  • Mouse

  • Rat

SPECIFICATIONS

Product Type

Recombinant Rabbit monoclonal primary

Product Name

Src Recombinant Rabbit Monoclonal Antibody [JF0947] (ET1702-03)

Immunogen

Recombinant protein

Host

Rabbit

Positive Control

HUVEC cell lysate, A549 cell lysate, PC-3M cell lysate, SH-SY5Y cell lysate, Hela, MCF-7, A431, human tonsil tissue, human breast carcinoma tissue, human kidney tissue, rat brain tissue, mouse kidney tissue, NIH/3T3.

Conjugation

Unconjugated

Clonality

Monoclonal

Clone Number

JF0947

PROPERTIES

Form

Liquid

Storage Condition

Store at +4C after thawing. Aliquot store at -20C or -80C. Avoid repeated freeze / thaw cycles.

Storage Buffer

1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Concentration

1 ug/ul

PURIFICATION

Protein A affinity purified.

MOLECULAR WEIGHT

60 kDa

Isotype

IgG

APPLICATION DILUTION

  • WB

  • 1:1,000-1:5,000

  • ICC/IF

  • 1:100-1:500

  • IHC-P

  • 1:50-1:200

  • FC

  • 1:50-1:100

TARGET

UNIPROT #

PROTEIN NAME

Src

SYNONYMS

ASV antibody; Avian sarcoma virus antibody; c SRC antibody; CDNA FLJ14219 fis clone NT2RP3003800 highly similar to Rattus norvegicus tyrosine protein kinase pp60 c src mRNA antibody; cSrc antibody; EC 2.7.10.2 antibody; Neuronal CSRC tyrosine specific protein kinase antibody; Neuronal SRC antibody; Oncogene SRC antibody; OTTHUMP00000174476 antibody; OTTHUMP00000174477 antibody; p60 Src antibody; p60-Src antibody; p60Src antibody; pp60c src antibody; pp60c-src antibody; pp60csrc antibody; Proto oncogene tyrosine protein kinase Src antibody; Proto-oncogene c-Src antibody; Proto-oncogene tyrosine-protein kinase Src antibody; Protooncogene SRC antibody; Protooncogene SRC Rous sarcoma antibody; Src antibody; SRC Oncogene antibody; SRC proto oncogene non receptor tyrosine kinase antibody; SRC_HUMAN antibody; SRC1 antibody; Tyrosine kinase pp60c src antibody; Tyrosine protein kinase SRC 1 antibody; Tyrosine protein kinase SRC1 antibody; v src avian sarcoma (Schmidt Ruppin A2) viral oncogene homolog antibody; V src sarcoma (Schmidt Ruppin A 2) viral oncogene homolog (avian) antibody; v src sarcoma (Schmidt Ruppin A 2) viral oncogene homolog avian antibody

SEQUENCE SIMILARITIES

Belongs to the protein kinase superfamily. Tyr protein kinase family. SRC subfamily.

TISSUE SPECIFICITY

Expressed ubiquitously. Platelets, neurons and osteoclasts express 5-fold to 200-fold higher levels than most other tissues.

POST-TRANSLATIONAL MODIFICATION

Myristoylated at Gly-2, and this is essential for targeting to membranes.; Dephosphorylated at Tyr-530 by PTPRJ (By similarity). Phosphorylated on Tyr-530 by c-Src kinase (CSK). The phosphorylated form is termed pp60c-src. Dephosphorylated by PTPRJ at Tyr-419. Normally maintained in an inactive conformation with the SH2 domain engaged with Tyr-530, the SH3 domain engaged with the SH2-kinase linker, and Tyr-419 dephosphorylated. Dephosphorylation of Tyr-530 as a result of protein tyrosine phosphatase (PTP) action disrupts the intramolecular interaction between the SH2 domain and Tyr-530, Tyr-419 can then become autophosphorylated, resulting in SRC activation. Phosphorylation of Tyr-530 by CSK allows this interaction to reform, resulting in SRC inactivation. CDK5-mediated phosphorylation at Ser-75 targets SRC to ubiquitin-dependent degradation and thus leads to cytoskeletal reorganization. Phosphorylated by PTK2/FAK1; this enhances kinase activity. Phosphorylated by PTK2B/PYK2; this enhances kinase activity.; S-nitrosylation is important for activation of its kinase activity.; Ubiquitinated in response to CDK5-mediated phosphorylation. Ubiquitination mediated by CBLC requires SRC autophosphorylation at Tyr-419 and may lead to lysosomal degradation.

SUBCELLULAR LOCATION

Cell membrane, Mitochondrion inner membrane, Nucleus, Cytoplasm.

FUNCTION

The major translational products of the Src gene family are membrane-associated tyrosine protein kinases that lack transmembrane and external amino acid sequences. By virtue of their common structural motifs, the Src family is composed of nine members in vertebrates, including c-Src, c-Yes, Fgr, Yrk, Fyn, Lyn, Hck, Lck and Blk. Src family kinases, which contain an amino-terminal cell membrane anchor followed by SH3 and SH2 domains, transduce signals that are involved in the control of a variety of cellular processes, including proliferation, differentiation, motility and adhesion. Src family members are normally maintained in an inactive state and can be activated transiently during cellular events such as mitosis. Different subcellular locations of Src family kinases may be important for the regulation of specific cellular processes, such as mitogenesis, cytoskeletal organization and membrane trafficking. c-Src (also designated pp60Src, Src p60 and proto-oncogene tyrosine protein kinase Src) is expressed in a broad range of tissue and cell types, although the highest levels of c-Src are detected in neuronal tissues and platelets. c-Src may play a role in events associated with both neuronal differentiation and maintenance of mature neuronal cell functions.

CITATIONS

  • Xiang, Bingyu et al.

    Transplantation of Menstrual Blood-Derived Mesenchymal Stem Cells Promotes the Repair of LPS-Induced Acute Lung Injury. | International Journal of Molecular Sciences [2017]

  • Pan, Jing et al.

    Pterostilbene, a bioactive component of blueberries, alleviates renal fibrosis in a severe mouse model of hyperuricemic nephropathy. | Biomedicine & Pharmacotherapy = Biomedecine & Pharmacotherapie [2019]