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78451
Angiogenesis Receptor Tyrosine Kinase Antibody Sampler Kit
Primary Antibodies
Antibody Sampler Kit
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Angiogenesis Receptor Tyrosine Kinase Antibody Sampler Kit #78451

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Simple Western™ analysis of lysates (0.1 mg/mL) from 3T3 cells using PDGF Receptor β (28E1) Rabbit mAb #3169. The virtual lane view (left) shows a single target band (as indicated) at 1:10 and 1:50 dilutions of primary antibody. The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:10 (blue line) and 1:50 (green line) dilutions of primary antibody. This experiment was performed under reducing conditions on the Jess™ Simple Western instrument from ProteinSimple, a BioTechne brand, using the 12-230 kDa separation module.
Confocal immunofluorescent analysis of fixed frozen mouse cerebellum labeled with PDGF Receptor β (28E1) Rabbit mAb (green, left), β3-Tubulin (E9F3E) Mouse mAb #45058 (red, right), and DAPI #4083 (blue, right).
Confocal immunofluorescent analysis of fixed frozen mouse cerebral cortex labeled with PDGF Receptor β (28E1) Rabbit mAb (green, left), β3-Tubulin (E9F3E) Mouse mAb #45058 (red, right), and DAPI #4083 (blue, right).
Western blot analysis of extracts from various cell lines using VEGF-A (E9X8Q) Rabbit mAb (upper) or GAPDH (D16H11) XP® Rabbit mAb #5174 (lower). Negative expression of VEGF-A protein in HuT 102 cells is consistent with the predicted expression pattern.
Flow cytometric analysis of A-204 cells using FGF Receptor 1 (D8E4) XP® Rabbit mAb (solid line) compared to concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype control #3900 (dashed line). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.
Phospho-VEGF Receptor 2 (Tyr1175) (19A10) Rabbit mAb specifically binds to phosphorylated VEGFR2, but not other phosphorylated tyrosine kinases. Western blot analysis of extracts from cells expressing different activated tyrosine kinase proteins, using Phospho-VEGF Receptor-2 (Tyr1175) (19A10) Rabbit mAb (upper) and Phospho-Tyrosine mAb (P-Tyr-100) #9411 (lower). CKR/PAE cells (lanes 12 and 13) express chimeric receptors containing human CSF-1 extracellular binding domain/mouse VEGF receptor-2 intracellular domain (7). CSF-1 stimulates phosphorylation of Tyr1175 of intracellular VEGF receptor-2 domain (lane 12), which was specifically detected by Phospho-VEGF Receptor-2 (Tyr1175) (19A10) Rabbit mAb.
Western blot analysis of extracts from NIH/3T3 cells, untreated or treated with PDGF (50 ng/ml for 5 minutes) and/or calf intestinal phosphatase (CIP) as indicated, using Phospho-PDGF Receptor β (Tyr751) (88H8) Mouse mAb.
Western blot analysis of extracts from various cell lines, using PDGF Receptor β (28E1) Rabbit mAb.
Western blot analysis of extracts from 293 cells, transfected with wild-type Tie2 (lane 1) or mock-transfected (lane 2), using Phospho-Tie2 (Tyr992) Antibody. Tie2 is constitutively active when transfected into 293 cells.
Immunoprecipitation of VEGF-A protein from U-87 MG cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is VEGF-A (E9X8Q) Rabbit mAb. Western blot analysis was performed using VEGF-A (E9X8Q) Rabbit mAb. Mouse Anti-rabbit IgG (Conformation Specific) (L27A9) mAb (HRP Conjugate) #5127 was used as a secondary antibody.
Western blot analysis of extracts from A-204 cells, untreated (-) or treated with heparin (1 μg/mL, 5 min) followed by the Human Basic Fibroblast Growth Factor (hFGF basic/FGF2) #61977 (100 ng/mL, 5 min, +), using Phospho-FGF Receptor 1 (Tyr653/654) (D4X3D) Rabbit mAb (upper) and FGF Receptor 1 (D8E4) XP® Rabbit mAb #9740 (lower).
After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO® is added and emits light during enzyme catalyzed decomposition.
Western blot analysis of extracts from HUVEC and HMVEC cells using Tie2 (D9D10) Rabbit mAb.
Western blot analysis of extracts from HUVE cells, HeLa cells and mouse heart using VEGF Receptor 2 (D5B1) Rabbit mAb for VEGFR2 expression (upper) and β-Actin (D6A8) Rabbit mAb (#8457) for loading control. (The 80 kDa bands represent partial degradation product).
Western blot analysis of extracts from A-204 (FGFR1 positive), KG-1a (FGFR1 oncogenic partner-FGFR1 fusion), A172 (FGFR1 low), and HT-29 (FGFR1 negative) cells using FGF Receptor 1 (D8E4) XP® Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower).
Western blot analysis of extracts from HUVE cells, untreated or stimulated with VEGF (50 ng/ml for 2 minutes), using Phospho-VEGF Receptor 2 (Tyr1175) (19A10) Rabbit mAb (upper) and VEGF Receptor 2 (55B11) Rabbit mAb #2479 (lower).
Immunohistochemical analysis of paraffin-embedded human colon carcinoma using PDGF Receptor β (28E1) Rabbit mAb.
Western blot analysis of extracts from Sf9 cells over expressing GST-human Tie2 kinase domain fusion proteins, wild-type (lane 1) or kinase-dead (lane 2), using Phospho-Tie2 (Tyr992) Antibody (upper) or Tie2 antibody (lower). The wild-type Tie2 kinase domain is constitutively phosphorylated when overexpressed in Sf9 cells. The molecular weight of GST-Tie2 fusion is approximately 65 kDa.
Immunoprecipitation of Phospho-FGF receptor 1 (Tyr653/654) protein from A-204 cell treated with heparin (1 μg/mL, 5 min) followed by the Human Basic Fibroblast Growth Factor (hFGF basic/FGF2) #61977 (100 ng/mL, 5 min). Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is Phospho-FGF Receptor 1(Tyr653/654) (D4X3D) Rabbit mAb. Western blot analysis was performed using Phospho-FGF Receptor 1(Tyr653/654) (D4X3D) Rabbit mAb as primary antibody. Anti-rabbit IgG, HRP-linked Antibody #7074 was used as secondary antibody.
Immunohistochemical analysis of paraffin-embedded human angiosarcoma using VEGF Receptor 2 (D5B1) Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded human breast carcinoma using FGF Receptor 1 (D8E4) XP® Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded human glioblastoma using PDGF Receptor β (28E1) Rabbit mAb.
Confocal immunofluorescent analysis of serum-starved A-204 cells, untreated (left), treated with heparin (1 μg/mL, 5 min) followed by the addition of Human Basic Fibroblast Growth Factor (hFGF basic/FGF2) #61977 (100 ng/mL, 5 min; center), or treated with heparin/FGF2 and post-processed with λ-phosphatase (2 hr; right), using Phospho-FGF Receptor 1 (Tyr653/654) (D4X3D) Rabbit mAb (upper, green) and FGF Receptor 1 (D8E4) XP® Rabbit mAb #9740 (lower, green). Samples were mounted in ProLong® Gold Antifade Reagent with DAPI #8961 (blue).
Immunohistochemical analysis of paraffin-embedded mouse liver using VEGF Receptor 2 (D5B1) Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded human kidney using FGF Receptor 1 (D8E4) XP® Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded U-87MG cells, showing membrane localization, using PDGF Receptor β (28E1) Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded HCC827 xenograft using VEGFR2 (D5B1) Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded human lung carcinoma using FGF Receptor 1 (D8E4) XP® Rabbit mAb.
Confocal immunofluorescent analysis of rat pancreas using VEGF Receptor 2 (D5B1) Rabbit mAb (green) and β-Catenin (L54E2) Mouse mAb (IF Preferred) #2677 (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
Immunohistochemical analysis of paraffin-embedded A-204 cell pellet (left, positive) or HT-29 cell pellet (right, negative) using FGF Receptor 1 (D8E4) XP® Rabbit mAb.
Confocal immunofluorescent analysis of NIH/3T3 cells, serum-starved (left) or PDGF-treated (right), using PDGF Receptor beta (28E1) Rabbit mAb (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
Flow cytometric analysis of fixed and permeabilized HeLa cells (blue, negative) and HUVEC cells (green, positive) using VEGF Receptor 2 (D5B1) Rabbit mAb (solid lines) or a concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed lines). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.
Confocal immunofluorescent analysis of A204 cells (positive, left), KG-1 cells (positive, middle) and A172 cells (weak expression, right) using FGF Receptor 1 (D8E4) XP® Rabbit mAb (green). Blue pseudocolor= DRAQ5® #4084 (fluorescent DNA dye).
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To Purchase # 78451
Cat. # Size Qty. Price
78451T		 1 Kit  (6 x 20 microliters)

Bulk & Custom Formulation

Product Includes Quantity Applications Reactivity MW(kDa) Isotype
VEGF-A (E9X8Q) Rabbit mAb 50661 20 µl
  • WB
  • IP
 H 16, 20, 23, 26 Rabbit IgG
VEGF Receptor 2 (D5B1) Rabbit mAb 9698 20 µl
  • WB
  • IP
  • IHC
  • IF
  • F
 H M R 210, 230 Rabbit IgG
Phospho-VEGF Receptor 2 (Tyr1175) (19A10) Rabbit mAb 2478 20 µl
  • WB
 H M 230 Rabbit IgG
PDGF Receptor β (28E1) Rabbit mAb 3169 20 µl
  • WB
  • IP
  • IHC
  • IF
 H M R 190 Rabbit IgG
Phospho-PDGF Receptor β (Tyr751) (88H8) Mouse mAb 3166 20 µl
  • WB
  • IP
 H M R 190 Mouse IgG2b
FGF Receptor 1 (D8E4) XP® Rabbit mAb 9740 20 µl
  • WB
  • IP
  • IHC
  • IF
  • F
 H M R Mk 92 , 120, 145 Rabbit IgG
Phospho-FGF Receptor 1 (Tyr653/654) (D4X3D) Rabbit mAb 52928 20 µl
  • WB
  • IP
  • IF
 H 120, 145 Rabbit IgG
Phospho-Tie2 (Tyr992) Antibody 4221 20 µl
  • WB
 H 160 Rabbit 
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl
  • WB
 Goat 
Tie2 (D9D10) Rabbit mAb 7403 20 µl
  • WB
 H 160 Rabbit IgG

Product Description

The Angiogenesis Receptor Tyrosine Kinase Antibody Sampler Kit provides an economical means of detecting the activation of VEGF receptor 2, FGF receptor 1, PDGF receptor beta, Tie2, and VEGF-A using phospho-specific and control antibodies. The kit includes enough antibodies to perform two western blot experiments with each primary antibody.

Background

Angiogenesis is defined as the physiological process by which new blood vessels are formed from pre-existing blood vessels. It is a critical process that enables specific physiological conditions in healthy adults, including development, skeletal muscle hypertrophy, and wound healing (1,2). Angiogenesis can be aberrantly activated to generate new blood vessels during pathological conditions such as cancer, neovascular disorders, and chronic inflammation. Angiogenesis is a complicated process regulated by multiple mechanisms and pathways (3). VEGFR2 is a member of the family of receptor tyrosine kinases (RTKs) mainly located in endothelial cells and is a major player in angiogenesis. VEGF-VEGFR2 ligand-receptor activation is the key signaling pathway for angiogenesis activation (4,5). Multiple non-VEGF RTK activations can replace VEGFR to promote angiogenesis (6,7), including PDGFR (9), FGFR (8), and Tie2 (10). These RTKs are targets for an antiangiogenic based disease treatment (11,12).

Pathways

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Limited Uses

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For Research Use Only. Not for Use in Diagnostic Procedures.
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