VEGF Receptor 1 Recombinant Rabbit Monoclonal Antibody [SY09-09]

Overview

Product Name

VEGF Receptor 1 Recombinant Rabbit Monoclonal Antibody [SY09-09] - BSA and Azide free

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic peptide within Human VEGF Receptor 1 aa 1-50 / 1,338.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IF-Cell, IF-Tissue, IHC-P, IP, FC

Target Molecular Weight

Predicted band size: 151 kDa

Positive Control

MCF7 cell lysate, human brain tissue lysate, mouse brain tissue lysate, rat brain tissue lysate, N2A, RH-35, SHG-44, mouse placenta tissue, mouse brain tissue, A431.

Conjugation

unconjugated

Clone Number

SY09-09

Reactivity Data

Tested

Verified (internally validated)

Published

Reported in literature (not internally validated)

Predicted

Predicted reactive (based on sequence homology)

Not recommended

Not recommended (failed internal validation)

	WB	IHC-P	FC(Intra)	IF-Cell	IP
human
mouse
rat

Product Features

Form

Liquid

Concentration

1mg/ml

Storage Instructions

Store at 2-8℃. Avoid freeze.

Storage Buffer

1*PBS (pH7.4).

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

Target

Function

Three cell membrane receptor tyrosine kinases, Flt (also designated VEGF-R1), Flk-1 (also designated VEGF-R2) and Flt-4, putatively involved in the growth of endothelial cells, are characterized by the presence of seven immunoglobulin-like sequences in their extracellular domain. These receptors exhibit high degrees of sequence relatedness to each other as well as lesser degrees of relatedness to the class III receptors including CSF-1/Fms, PDGR, SLFR/Kit and Flt-3/Flk-2. Two members of this receptor class, Flt-1 and Flk-1, have been shown to represent high affinity receptors for vascular endothelial growth factors (VEGFs). On the basis of structural similarity to Flt and Flk-1, it has been speculated that Flt-4 might represent a third receptor for either VEGF or a VEGF-related ligand.

Background References

1. Zhou W et al. Fibroblast growth factor receptor 1 promotes MG63 cell proliferation and is associated with increased expression of cyclin-dependent kinase 1 in osteosarcoma. Mol Med Rep 13:713-9 (2016).

2. Liu X et al. Impaired VEGF Signaling in Lungs with Hypoplastic Esophageal Atresia and Effects on Branching Morphogenesis. Cell Physiol Biochem 39:385-94 (2016).

Sequence Similarity

Belongs to the protein kinase superfamily. Tyr protein kinase family. CSF-1/PDGF receptor subfamily.

Tissue Specificity

Detected in normal lung, but also in placenta, liver, kidney, heart and brain tissues. Specifically expressed in most of the vascular endothelial cells, and also expressed in peripheral blood monocytes. Isoform 2 is strongly expressed in placenta. Isoform 3 is expressed in corneal epithelial cells (at protein level). Isoform 3 is expressed in vascular smooth muscle cells (VSMC).

Post-translational Modification

N-glycosylated.; Ubiquitinated after VEGFA-mediated autophosphorylation, leading to proteolytic degradation.; Autophosphorylated on tyrosine residues upon ligand binding. Autophosphorylation occurs in trans, i.e. one subunit of the dimeric receptor phosphorylates tyrosine residues on the other subunit. Phosphorylation at Tyr-1169 is important for interaction with PLCG. Phosphorylation at Tyr-1213 is important for interaction with PIK3R1, PTPN11, GRB2, and PLCG. Phosphorylation at Tyr-1333 is important for endocytosis and for interaction with CBL, NCK1 and CRK. Is probably dephosphorylated by PTPRB.

Subcellular Location

Cell membrane, Endosome, Secreted, Cytoplasm.

UNIPROT

SwissProt: P17948 Human

SwissProt: P35969 Mouse

SwissProt: P53767 Rat

Synonyms

EC 2.7.10.1 antibody

FLT 1 antibody

FLT antibody

Flt-1 antibody

FLT1 antibody

Fms like tyrosine kinase 1 antibody

Fms related tyrosine kinase 1 antibody

Fms related tyrosine kinase 1 (vascular endothelial growth factor/vascular permeability factor receptor) antibody

Fms related tyrosine kinase 1 vascular endothelial growth factor/vascular permeability factor receptor antibody

Fms-like tyrosine kinase 1 antibody

Expand

Images

This data was developed using ET1605-11, the same antibody clone in a different buffer formulation.
Western blot analysis of VEGF Receptor 1 on different lysates with Rabbit anti-VEGF Receptor 1 antibody (ET1605-11) at 1/5,000 dilution.

Lane 1: MCF7 cell lysate (20 µg/Lane)
Lane 2: Human brain tissue lysate (20 µg/Lane)
Lane 3: Mouse brain tissue lysate (20 µg/Lane)
Lane 4: Rat brain tissue lysate (20 µg/Lane)

Predicted band size: 151 kDa
Observed band size: 151 kDa

Exposure time: 24 seconds;

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 (ET1605-11) at 1/5,000 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.
☑ Knockdown (KD)

This data was developed using ET1605-11, the same antibody clone in a different buffer formulation.
Western blot analysis of VEGF Receptor 1 on different lysates with Rabbit anti-VEGF Receptor 1 antibody (ET1605-11) at 1/1,000 dilution.

Lane 1: MCF7-si NT cell lysate (10 µg/Lane)
Lane 2: MCF7-si VEGF Receptor 1 cell lysate (10 µg/Lane)

Predicted band size: 151 kDa
Observed band size: 151 kDa

Exposure time: 25 seconds;

4-20% SDS-PAGE gel.

ET1605-11 was shown to specifically react with VEGF Receptor 1 in Hela-si NT cells. Weakened band was observed when Hela-si VEGF Receptor 1 sample was tested. Hela-si NT and Hela-si VEGF Receptor 1 samples were subjected to SDS-PAGE. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM in TBST for 1 hour at room temperature. The primary antibody (ET1605-11, 1/1,000) and Loading control antibody (Rabbit anti-GAPDH, ET1601-4, 1/10,000) were used in 5% BSA at room temperature for 2 hours. Goat Anti-rabbit IgG-HRP Secondary Antibody (HA1001) at 1:100,000 dilution was used for 1 hour at room temperature.
This data was developed using ET1605-11, the same antibody clone in a different buffer formulation.
VEGF Receptor 1 was immunoprecipitated in 0.2mg MCF7 cell lysate with ET1605-11 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using ET1605-11 at 1/5,000 dilution. Anti-Rabbit IgG for IP, AlpSdAbs® VHH(HRP) (025-100-005) at 1/5,000 dilution was used for 1 hour at room temperature.

Lane 1: MCF7 cell lysate (input)
Lane 2: Rabbit IgG instead of ET1605-11 in MCF7 cell lysate
Lane 3: ET1605-11 IP in MCF7 cell lysate

Blocking/Dilution buffer: 5% NFDM/TBST
Exposure time: 24 seconds
This data was developed using ET1605-11, the same antibody clone in a different buffer formulation.
ICC staining of VEGF Receptor 1 in N2A cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 10% negative goat serum for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1605-11, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 conjugate-Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
This data was developed using ET1605-11, the same antibody clone in a different buffer formulation.
ICC staining of VEGF Receptor 1 in RH-35 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 10% negative goat serum for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1605-11, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 conjugate-Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
This data was developed using ET1605-11, the same antibody clone in a different buffer formulation.
ICC staining of VEGF Receptor 1 in SHG-44 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 10% negative goat serum for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1605-11, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 conjugate-Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
This data was developed using ET1605-11, the same antibody clone in a different buffer formulation.
Application: Immunohistochemistry (IHC-P)

Species: Human
Tissue: Gastric carcinoma
Sample: Paraffin-embedded section

Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95℃.

Wash buffer: 1× TBST
Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.
Primary antibody: ET1605-11, 1/5,000, 1 hour at room temperature.
Secondary antibody: HA1119, 20 minutes at room temperature.
This data was developed using ET1605-11, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded mouse placenta tissue using anti-VEGF Receptor 1 antibody. 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 5% BSA for 30 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (ET1605-11, 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.
This data was developed using ET1605-11, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded mouse brain tissue using anti-VEGF Receptor 1 antibody. 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 5% BSA for 30 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (ET1605-11, 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.
This data was developed using ET1605-11, the same antibody clone in a different buffer formulation.
Flow cytometric analysis of VEGF Receptor 1 was done on A431 cells. The cells were fixed, permeabilized and stained with the primary antibody (ET1605-11, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor®488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).