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PEDF Recombinant Rabbit Monoclonal Antibody [PSH28-40]

Usd: 385

Specification

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Catalog# HA724781

PEDF Recombinant Rabbit Monoclonal Antibody [PSH28-40]

Application

  • WB

  • FC(Intra)

  • IP

Reactivity

  • Human

  • Mouse

  • Rat

  • Pig

BSA and Azide free
Conjugation

  • unconjugated

Overview

Product Name

PEDF Recombinant Rabbit Monoclonal Antibody [PSH28-40]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Recombinant protein within human PEDF aa 1-418.

Species Reactivity

Human, Mouse, Rat, Pig

Validated Applications

WB, FC(Intra), IP

Target Molecular Weight

Predicted band size: 46 kDa

Positive Control

K-562 (Human chronic myelogenous leukemia cell) cell lysate, Y79 (Human retinoblastoma cell) cell lysate, Hep G2 (Human hepatocellular carcinoma cell) cell lysates, NIH/3T3 (Mouse fibroblast) cell lysate, 3T3-L1 (Mouse embryonic fibroblast) cell lysate, C2C12 (Mouse myoblast) cell lysate, NRK (Rat kidney cell) cell lysate, Mouse liver tissue lysate, Mouse brain tissue lysate, Mouse stomach tissue lysate, Rat liver tissue lysate, Rat brain tissue lysate, Human serum lysate, Mouse plasma lysate, Rat plasma lysate, Pig liver tissue lysate, Pig brain tissue lysate.

Conjugation

unconjugated

Clone Number

PSH28-40

Reactivity Data

Tested Tested Tested
Verified (internally validated)
Published Tested
Reported in literature (not internally validated)
Predicted Tested
Predicted reactive (based on sequence homology)
Not recommended Tested
Not recommended (failed internal validation)
WB IHC-P FC(Intra) IF-Cell IP
human
Tested Tested
Not Recommended
Not Recommended
Not Recommended
Tested Tested
mouse
Tested Tested
Not Recommended
Tested Tested
Not Recommended
 
rat
Tested Tested
Not Recommended
Not Recommended
Not Recommended
 
pig
Tested Tested
 
 
 
 

Product Features

Form

Liquid

Concentration

0.5 mg/mL.(The concentration of this product may be batch-dependent)

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.

Storage Buffer

1*PBS (pH7.4), 0.1% BSA, 40% Glycerol, 0.2% Proclean 950.

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:2,500

  • FC(Intra)

  • 1:500

  • IP

  • 1-2μg/sample

Target

Function

Pigment epithelium-derived factor (PEDF) also known as serpin F1 (SERPINF1), is a multifunctional secreted protein that has anti-angiogenic, anti-tumorigenic, and neurotrophic functions. Found in vertebrates, this 50 kDa protein is being researched as a therapeutic candidate for treatment of such conditions as choroidal neovascularization, heart disease, and cancer.[5] In humans, pigment epithelium-derived factor is encoded by the SERPINF1 gene. PEDF has a variety of functions including antiangiogenic, antitumorigenic, and neurotrophic properties. Endothelial cell migration is inhibited by PEDF. PEDF suppresses retinal neovascularization and endothelial cell proliferation. The antiangiogenic residues 24-57 were shown to be sufficient at inhibiting angiogenesis. PEDF is also responsible for apoptosis of endothelial cells either through the p38 MAPK pathway or through the FAS/FASL pathway Antiangiogenic function is also conferred by PEDF through inhibition of both VEGFR-1 and VEGFR-2. The antitumorigenic effects of PEDF are not only due to inhibition of supporting vasculature, but also due to effects on the cancer cells themselves. PEDF was shown to inhibit cancer cell proliferation and increase apoptosis via the FAS/FASL pathway. VEGF expression by cancer cells is inhibited by PEDF. PEDF also displays neurotrophic functions. Retinoblastoma cells differentiate into neurons due to the presence of PEDF. Expression of PEDF in the human retina is found at 7.4 weeks of gestation, suggesting it may play a role in retinal neuron differentiation.

Background References

1. Carpino G et al. Thrombospondin 1 and 2 along with PEDF inhibit angiogenesis and promote lymphangiogenesis in intrahepatic cholangiocarcinoma. J Hepatol. 2021 Dec

2. Ma B et al. Pigment epithelium-derived factor (PEDF) plays anti-inflammatory roles in the pathogenesis of dry eye disease. Ocul Surf. 2021 Apr

Subcellular Location

Secreted, Melanosome.

UNIPROT

SwissProt: P36955 Human

SwissProt: P97298 Mouse

Entrez Gene: 287526 Rat

Synonyms

Cell proliferation-inducing gene 35 protein antibody

EPC 1 antibody

EPC-1 antibody

EPC1 antibody

OI12 antibody

OI6 antibody

PEDF antibody

PEDF_HUMAN antibody

PIG 35 antibody

PIG35 antibody

Expand

Images

  • <span style="font-weight: bold;">☑ Relative expression (RE)</span><br /><br />Western blot analysis of PEDF on different lysates with Rabbit anti-PEDF antibody (<a href="/products/HA724781" style="font-weight: bold;text-decoration: underline;">HA724781</a>) at 1/2,500 dilution. <br /> <br />Lane 1: K-562 (Human chronic myelogenous leukemia cell) cell lysate <br />Lane 2: Y79 (Human retinoblastoma cell) cell lysate <br />Lane 3: MCF7 (Human breast cancer cell) cell lysate <br /> <br />Lysates/proteins at 15 µg/Lane. <br />Exposure time: 2 minutes 6 seconds; ECL: K1801 <br /> <br />MCF7 is a negative control (PMID: 16740777). <br /> <br />Blocking: 5% NFDM/TBST, 1 hour at room temperature <br />Primary antibody: <a href="/products/HA724781" style="font-weight: bold;text-decoration: underline;">HA724781</a>, 1/2,500 in primary antibody dilution buffer (<a href="/products/K1803" style="font-weight: bold;text-decoration: underline;">K1803</a>), overnight at 4 °C <br />Secondary antibody: Goat anti-Rabbit IgG-HRP (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature <br /> <br />Predicted band size: 46 kDa <br />Observed band size: 50 kDa

    ☑ Relative expression (RE)

    Western blot analysis of PEDF on different lysates with Rabbit anti-PEDF antibody (HA724781) at 1/2,500 dilution.

    Lane 1: K-562 (Human chronic myelogenous leukemia cell) cell lysate
    Lane 2: Y79 (Human retinoblastoma cell) cell lysate
    Lane 3: MCF7 (Human breast cancer cell) cell lysate

    Lysates/proteins at 15 µg/Lane.
    Exposure time: 2 minutes 6 seconds; ECL: K1801

    MCF7 is a negative control (PMID: 16740777).

    Blocking: 5% NFDM/TBST, 1 hour at room temperature
    Primary antibody: HA724781, 1/2,500 in primary antibody dilution buffer (K1803), overnight at 4 °C
    Secondary antibody: Goat anti-Rabbit IgG-HRP (HA1001), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature

    Predicted band size: 46 kDa
    Observed band size: 50 kDa

  • Western blot analysis of PEDF on Hep G2 (Human hepatocellular carcinoma cell) cell lysate with Rabbit anti-PEDF antibody (<a href="/products/HA724781" style="font-weight: bold;text-decoration: underline;">HA724781</a>) at 1/2,500 dilution. <br /> <br />Lysates/proteins at 15 µg/Lane. <br />Exposure time: 10 seconds; ECL: K1801 <br /> <br />Blocking: 5% NFDM/TBST, 1 hour at room temperature <br />Primary antibody: <a href="/products/HA724781" style="font-weight: bold;text-decoration: underline;">HA724781</a>, 1/2,500 in primary antibody dilution buffer (<a href="/products/K1803" style="font-weight: bold;text-decoration: underline;">K1803</a>), overnight at 4 °C <br />Secondary antibody: Goat anti-Rabbit IgG-HRP (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature <br /> <br />Predicted band size: 46 kDa <br />Observed band size: 50 kDa

    Western blot analysis of PEDF on Hep G2 (Human hepatocellular carcinoma cell) cell lysate with Rabbit anti-PEDF antibody (HA724781) at 1/2,500 dilution.

    Lysates/proteins at 15 µg/Lane.
    Exposure time: 10 seconds; ECL: K1801

    Blocking: 5% NFDM/TBST, 1 hour at room temperature
    Primary antibody: HA724781, 1/2,500 in primary antibody dilution buffer (K1803), overnight at 4 °C
    Secondary antibody: Goat anti-Rabbit IgG-HRP (HA1001), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature

    Predicted band size: 46 kDa
    Observed band size: 50 kDa

  • Western blot analysis of PEDF on different lysates with Rabbit anti-PEDF antibody (<a href="/products/HA724781" style="font-weight: bold;text-decoration: underline;">HA724781</a>) at 1/2,500 dilution. <br /> <br />Lane 1: NIH/3T3 (Mouse fibroblast) cell lysate <br />Lane 2: 3T3-L1 (Mouse embryonic fibroblast) cell lysate <br />Lane 3: C2C12 (Mouse myoblast) cell lysate <br />Lane 4: NRK (Rat kidney cell) cell lysate <br /> <br />Lysates/proteins at 20 µg/Lane. <br />Exposure time: 18 seconds; ECL: K1801 <br /> <br />Blocking: 5% NFDM/TBST, 1 hour at room temperature <br />Primary antibody: <a href="/products/HA724781" style="font-weight: bold;text-decoration: underline;">HA724781</a>, 1/2,500 in primary antibody dilution buffer (<a href="/products/K1803" style="font-weight: bold;text-decoration: underline;">K1803</a>), overnight at 4 °C <br />Secondary antibody: Goat anti-Rabbit IgG-HRP (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature <br /> <br />Predicted band size: 46 kDa <br />Observed band size: 50 kDa

    Western blot analysis of PEDF on different lysates with Rabbit anti-PEDF antibody (HA724781) at 1/2,500 dilution.

    Lane 1: NIH/3T3 (Mouse fibroblast) cell lysate
    Lane 2: 3T3-L1 (Mouse embryonic fibroblast) cell lysate
    Lane 3: C2C12 (Mouse myoblast) cell lysate
    Lane 4: NRK (Rat kidney cell) cell lysate

    Lysates/proteins at 20 µg/Lane.
    Exposure time: 18 seconds; ECL: K1801

    Blocking: 5% NFDM/TBST, 1 hour at room temperature
    Primary antibody: HA724781, 1/2,500 in primary antibody dilution buffer (K1803), overnight at 4 °C
    Secondary antibody: Goat anti-Rabbit IgG-HRP (HA1001), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature

    Predicted band size: 46 kDa
    Observed band size: 50 kDa

  • Western blot analysis of PEDF on different lysates with Rabbit anti-PEDF antibody (<a href="/products/HA724781" style="font-weight: bold;text-decoration: underline;">HA724781</a>) at 1/2,500 dilution. <br /> <br />Lane 1: Mouse liver tissue lysate <br />Lane 2: Mouse brain tissue lysate <br />Lane 3: Mouse stomach tissue lysate <br />Lane 4: Rat liver tissue lysate <br />Lane 5: Rat brain tissue lysate <br /> <br />Lysates/proteins at 20 µg/Lane. <br />Exposure time: 1 minute 2 seconds; ECL: K1801 <br /> <br />Blocking: 5% NFDM/TBST, 1 hour at room temperature <br />Primary antibody: <a href="/products/HA724781" style="font-weight: bold;text-decoration: underline;">HA724781</a>, 1/2,500 in primary antibody dilution buffer (<a href="/products/K1803" style="font-weight: bold;text-decoration: underline;">K1803</a>), overnight at 4 °C <br />Secondary antibody: Goat anti-Rabbit IgG-HRP (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature <br /> <br />Predicted band size: 46 kDa <br />Observed band size: 50 kDa

    Western blot analysis of PEDF on different lysates with Rabbit anti-PEDF antibody (HA724781) at 1/2,500 dilution.

    Lane 1: Mouse liver tissue lysate
    Lane 2: Mouse brain tissue lysate
    Lane 3: Mouse stomach tissue lysate
    Lane 4: Rat liver tissue lysate
    Lane 5: Rat brain tissue lysate

    Lysates/proteins at 20 µg/Lane.
    Exposure time: 1 minute 2 seconds; ECL: K1801

    Blocking: 5% NFDM/TBST, 1 hour at room temperature
    Primary antibody: HA724781, 1/2,500 in primary antibody dilution buffer (K1803), overnight at 4 °C
    Secondary antibody: Goat anti-Rabbit IgG-HRP (HA1001), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature

    Predicted band size: 46 kDa
    Observed band size: 50 kDa

  • Western blot analysis of PEDF on different lysates with Rabbit anti-PEDF antibody (<a href="/products/HA724781" style="font-weight: bold;text-decoration: underline;">HA724781</a>) at 1/2,500 dilution. <br /> <br />Lane 1: Human serum lysate <br />Lane 2: Mouse plasma lysate <br />Lane 3: Rat plasma lysate <br /> <br />Lysates/proteins at 10 µg/Lane. <br />Exposure time: 18 seconds; ECL: K1801 <br /> <br />Blocking: 5% NFDM/TBST, 1 hour at room temperature <br />Primary antibody: <a href="/products/HA724781" style="font-weight: bold;text-decoration: underline;">HA724781</a>, 1/2,500 in primary antibody dilution buffer (<a href="/products/K1803" style="font-weight: bold;text-decoration: underline;">K1803</a>), overnight at 4 °C <br />Secondary antibody: Goat anti-Rabbit IgG-HRP (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature <br /> <br />Predicted band size: 46 kDa <br />Observed band size: 50 kDa

    Western blot analysis of PEDF on different lysates with Rabbit anti-PEDF antibody (HA724781) at 1/2,500 dilution.

    Lane 1: Human serum lysate
    Lane 2: Mouse plasma lysate
    Lane 3: Rat plasma lysate

    Lysates/proteins at 10 µg/Lane.
    Exposure time: 18 seconds; ECL: K1801

    Blocking: 5% NFDM/TBST, 1 hour at room temperature
    Primary antibody: HA724781, 1/2,500 in primary antibody dilution buffer (K1803), overnight at 4 °C
    Secondary antibody: Goat anti-Rabbit IgG-HRP (HA1001), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature

    Predicted band size: 46 kDa
    Observed band size: 50 kDa

  • Western blot analysis of PEDF on different lysates with Rabbit anti-PEDF antibody (<a href="/products/HA724781" style="font-weight: bold;text-decoration: underline;">HA724781</a>) at 1/2,500 dilution. <br /> <br />Lane 1: Pig liver tissue lysate <br />Lane 2: Pig brain tissue lysate <br /> <br />Lysates/proteins at 20 µg/Lane. <br />Exposure time: 1 minute 2 seconds; ECL: K1801 <br /> <br />Blocking: 5% NFDM/TBST, 1 hour at room temperature <br />Primary antibody: <a href="/products/HA724781" style="font-weight: bold;text-decoration: underline;">HA724781</a>, 1/2,500 in primary antibody dilution buffer (<a href="/products/K1803" style="font-weight: bold;text-decoration: underline;">K1803</a>), overnight at 4 °C <br />Secondary antibody: Goat anti-Rabbit IgG-HRP (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature <br /> <br />Predicted band size: 46 kDa <br />Observed band size: 50 kDa

    Western blot analysis of PEDF on different lysates with Rabbit anti-PEDF antibody (HA724781) at 1/2,500 dilution.

    Lane 1: Pig liver tissue lysate
    Lane 2: Pig brain tissue lysate

    Lysates/proteins at 20 µg/Lane.
    Exposure time: 1 minute 2 seconds; ECL: K1801

    Blocking: 5% NFDM/TBST, 1 hour at room temperature
    Primary antibody: HA724781, 1/2,500 in primary antibody dilution buffer (K1803), overnight at 4 °C
    Secondary antibody: Goat anti-Rabbit IgG-HRP (HA1001), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature

    Predicted band size: 46 kDa
    Observed band size: 50 kDa

  • Application: Flow Cytometry (Intra) <br /> <br />Species: Mouse <br />Sample: NIH/3T3 (Mouse fibroblasts) <br /> <br />Fixation: 4% Paraformaldehyde, 15 minutes at room temperature. <br />Permeabilization: 0.1% Tween-20, 15 minutes at room temperature. <br />Blocking: 1% BSA + 10% normal goat serum, 15 minutes at room temperature. <br />Antibody dilution buffer: 1x PBS. <br /> <br />Primary antibody: <a href="/products/HA724781" style="font-weight: bold;text-decoration: underline;">HA724781</a> (1/500, Red) compared with Rabbit IgG Isotype Control (<a href="/products/HA722127" style="font-weight: bold;text-decoration: underline;">HA722127</a>, Green), 15 minutes at room temperature. <br />Secondary antibody: Goat Anti-Rabbit IgG (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>), 15 minutes at room temperature.

    Application: Flow Cytometry (Intra)

    Species: Mouse
    Sample: NIH/3T3 (Mouse fibroblasts)

    Fixation: 4% Paraformaldehyde, 15 minutes at room temperature.
    Permeabilization: 0.1% Tween-20, 15 minutes at room temperature.
    Blocking: 1% BSA + 10% normal goat serum, 15 minutes at room temperature.
    Antibody dilution buffer: 1x PBS.

    Primary antibody: HA724781 (1/500, Red) compared with Rabbit IgG Isotype Control (HA722127, Green), 15 minutes at room temperature.
    Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 15 minutes at room temperature.

  • Immunoprecipitation (IP) <br /> <br />PEDF was immunoprecipitated in 0.2 mg Hep G2 (Human hepatocellular carcinoma cell) cell lysate with <a href="/products/HA724781" style="font-weight: bold;text-decoration: underline;">HA724781</a> at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using <a href="/products/HA724781" style="font-weight: bold;text-decoration: underline;">HA724781</a> at 1/2,500 dilution. Anti-Rabbit IgG for IP, AlpSdAbs&reg; VHH(HRP) (<a href="/products/025-100-005" style="font-weight: bold;text-decoration: underline;">025-100-005</a>) at 1/5,000 dilution was used for 1 hour at room temperature. <br /> <br />Lane 1: Hep G2 cell lysate (input) <br />Lane 2: <a href="/products/HA724781" style="font-weight: bold;text-decoration: underline;">HA724781</a> IP in Hep G2 cell lysate <br />Lane 3: Rabbit IgG instead of <a href="/products/HA724781" style="font-weight: bold;text-decoration: underline;">HA724781</a> in Hep G2 cell lysate <br /> <br />Exposure time: 12 seconds <br />Blocking: 5% NFDM/TBST, 1 hour at room temperature <br /> <br />Primary dilution: <a href="/products/HA724781" style="font-weight: bold;text-decoration: underline;">HA724781</a>, 1/2,500 in primary antibody dilution buffer (<a href="/products/K1803" style="font-weight: bold;text-decoration: underline;">K1803</a>), 2 hours at room temperature <br /> <br />Predicted band size: 46 kDa <br />Observed band size: 50 kDa

    Immunoprecipitation (IP)

    PEDF was immunoprecipitated in 0.2 mg Hep G2 (Human hepatocellular carcinoma cell) cell lysate with HA724781 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA724781 at 1/2,500 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: Hep G2 cell lysate (input)
    Lane 2: HA724781 IP in Hep G2 cell lysate
    Lane 3: Rabbit IgG instead of HA724781 in Hep G2 cell lysate

    Exposure time: 12 seconds
    Blocking: 5% NFDM/TBST, 1 hour at room temperature

    Primary dilution: HA724781, 1/2,500 in primary antibody dilution buffer (K1803), 2 hours at room temperature

    Predicted band size: 46 kDa
    Observed band size: 50 kDa

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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