Anti-VEGF Receptor 1 antibody [Y103]

• IP

Lab

Immunoprecipitation - Anti-VEGF Receptor 1 antibody [Y103] (AB32152)

VEGF Receptor 1 was immunoprecipitated from 0.35 mg mouse brain lysate 10μg with ab32152 at 1 : 30 dilution (2μg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab32152 1 : 1000 dilution (2 μg/ml). VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1 : 1000 dilution.

Lane 1 : Mouse brain lysate 10μg.

Lane 2 : ab32152 IP in mouse brain lysate.

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab32152 in mouse brain lysate.

Blocking and dilution buffer and concentration : 5% NFDM/TBST.

Exposure time : 1 second.

All lanes:

Immunoprecipitation - Anti-VEGF Receptor 1 antibody [Y103] (ab32152)

Predicted band size: 151 kDa

false

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-VEGF Receptor 1 antibody [Y103] (AB32152)

Immunohistochemical analysis of formalin fixed paraffin embedded human placenta labelling VEGF Receptor 1 with ab32152 at a concentration of 0.5µg/ml. The immunostaining was performed on a Ventana DISCOVERY ULTRA (Roche Tissue Diagnostics) instrument with a OptiView DAB IHC Detection Kit. Heat mediated antigen retrieval was performed with DISCOVERY cell conditioning solution (CC1) 100°C, pH8.5 for 32mins. ab32152 anti-VEGF Receptor 1 antibody [Y103] was incubated for 16mins at 37°C. Sections were counterstained with Hematoxylin II. Image inset shows absence of staining in secondary antibody only control.

Customers are encouraged to optimise antigen retrieval conditions, antibody concentration, incubation times and temperature for best results in their own IHC assay workflow (automated and manual).

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-VEGF Receptor 1 antibody [Y103] (AB32152)

Immunohistochemical analysis of paraffin-embedded Human gastric carcinoma tissue labeling VEGF with ab32152, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Cytoplasmic staining on human gastric carcinoma. Counterstained with Hematoxylin. Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-VEGF Receptor 1 antibody [Y103] (AB32152)

Immunohistochemical analysis of formalin fixed paraffin embedded human stomach adenocarcinoma labelling VEGF Receptor 1 with ab32152 at a concentration of 0.5µg/ml. The immunostaining was performed on a Ventana DISCOVERY ULTRA (Roche Tissue Diagnostics) instrument with a OptiView DAB IHC Detection Kit. Heat mediated antigen retrieval was performed with DISCOVERY cell conditioning solution (CC1) 100°C, pH8.5 for 32mins. ab32152 anti-VEGF Receptor 1 antibody [Y103] was incubated for 16mins at 37°C. Sections were counterstained with Hematoxylin II. Image inset shows absence of staining in secondary antibody only control.

Customers are encouraged to optimise antigen retrieval conditions, antibody concentration, incubation times and temperature for best results in their own IHC assay workflow (automated and manual).

• IHC-P

AbReview77520****

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-VEGF Receptor 1 antibody [Y103] (AB32152)

Immunohistochemistry analysis (Formalin/PFA-fixed paraffin-embedded sections) of paraformaldehyde fixed, 1% TritonX-100 in PBS permeabilized human Metastatic melanoma tissue. Stained with ab32152 at 1/100 dilution. Secondary antibody used was HRP Donkey Jackson Immuno Anti-rabbit at 1/500 dilution. Blocking was done with 3% donkey serum for 30 minutes at 25°C. The sample was incubated with the primary antibody and 1% donkey serum for 1 hour at 25°C. Antigen retrieval method was heat mediated Tris-EDTA

This image is courtesy of an anonymous Abreview

• IHC-P

PubMed

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-VEGF Receptor 1 antibody [Y103] (AB32152)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human abdominal aortic aneurysm (AAA) wall tissue sections labeling VEGF Receptor 1 with ab32152 at 1/100 dilution.

Resected aortic tissues were immersed in 10% neutral buffered formalin for at least 24 h for immunohistochemical staining. Tissue sample was embedded in paraffin; 4 μm sections were cut and mounted onto MAS-coated slides. The sections were deparaffinized, dehydrated, and boiled in a pressure cooker in 0.01 M citric acid buffer (pH 6.0) for 20 min. The sections were washed with phosphate-buffered saline and incubated with 3% H₂O₂ in absolute methanol for 5 min to inhibit any endogenous peroxidase activity. Sections were preincubated with 3% normal goat serum for 20 min to minimize nonspecific binding to VEGF Receptor 1, and incubated with ab32152 at 4°C overnight in a moist chamber. The section was washed with phosphate-buffered saline and then incubated with the appropriate secondary antibody for 30 min at room temperature. Staining was visualized with Vector DAB, and tissue section was then counterstained with hematoxylin.

Image from Sano M et al. PLoS ONE. 2014 Mar 20; 9(3). Fig 2E. DOI 10.1371/journal.pone.0089830. Lymphangiogenesis and Angiogenesis in Abdominal Aortic Aneurysm. e89830.

• WB

Lab

Western blot - Anti-VEGF Receptor 1 antibody [Y103] (AB32152)

Blocking and diluting buffer and concentration : 5% NFDM/TBST. Lysates were not boiled.

Lanes 1 - 2:

Western blot - Anti-VEGF Receptor 1 antibody [Y103] (ab32152) at 1/1000 dilution

Lanes 1 - 2:

Western blot - Anti-VEGF Receptor 1 antibody [Y103] - BSA and Azide free (<a href='/en-us/products/primary-antibodies/vegf-receptor-1-antibody-y103-bsa-and-azide-free-ab210608'>ab210608</a>)

Lanes 1 - 2:

Western blot - Anti-VEGF Receptor 1 antibody [Y103] - Low endotoxin, Azide free (<a href='/en-us/products/primary-antibodies/vegf-receptor-1-antibody-y103-low-endotoxin-azide-free-ab184784'>ab184784</a>)

Lanes 1 - 2:

Western blot - Anti-6X His tag® antibody [EPR20547] - ChIP Grade (<a href='/en-us/products/primary-antibodies/6x-his-tag-antibody-epr20547-chip-grade-ab213204'>ab213204</a>) at 1/5000 dilution

Lane 1:

HEK-293T (human embryonic kidney epithelial cell) cells transfected with an empty vector containing a His tag whole cell lysate at 20 µg

Lane 2:

HEK-293T (human embryonic kidney epithelial cell) cells transfected with sFlt1 (P17948-2) expression vector containing a His -tag whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 77 kDa

false

Exposure time: 5s

• WB

Supplier Data

Western blot - Anti-VEGF Receptor 1 antibody [Y103] (AB32152)

Blocking/Diluting buffer and concentration : 5% NFDM/TBST.

All lanes:

Western blot - Anti-VEGF Receptor 1 antibody [Y103] (ab32152) at 1/1000 dilution

Lane 1:

Mouse brain lysates at 15 µg

Lane 2:

Human brain lysates at 15 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 151 kDa

Observed band size: 180 kDa

false

Exposure time: 40s

• WB

Lab

Western blot - Anti-VEGF Receptor 1 antibody [Y103] (AB32152)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/1000,000 dilution.

In Western blot, Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204) 1 : 5,000 dilution used as loading control.

All lanes:

Western blot - Anti-VEGF Receptor 1 antibody [Y103] (ab32152) at 1/1000 dilution

Lane 1:

HEK-293T (human embryonic kidney epithelial cell) cells transfected with an empty vector contain a His-tag whole cell lysate at 20 µg

Lane 2:

HEK-293T cells transfected with a human VEGF Receptor 1 expression vector containing a His-tag whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 180 kDa

false

Exposure time: 1s

• WB

Unknown

Western blot - Anti-VEGF Receptor 1 antibody [Y103] (AB32152)

All lanes:

Western blot - Anti-VEGF Receptor 1 antibody [Y103] (ab32152) at 1/10000 dilution

All lanes:

mouse brain tissue

Predicted band size: 151 kDa

Observed band size: 180 kDa

false

• WB

Lab

Western blot - Anti-VEGF Receptor 1 antibody [Y103] (AB32152)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/1000,000 dilution.

All lanes:

Western blot - Anti-VEGF Receptor 1 antibody [Y103] (ab32152) at 1/1000 dilution

Lane 1:

Mouse placenta tissue lysate at 20 µg

Lane 2:

Rat placenta tissue lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 180 kDa

false

Exposure time: 10s