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FGFR2 Recombinant Rabbit Monoclonal Antibody [PSH26-90] - BSA and Azide free

Usd: 649

Specification

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Catalog# HA752052

FGFR2 Recombinant Rabbit Monoclonal Antibody [PSH26-90] - BSA and Azide free

Application

  • WB

  • FC

  • IP

Reactivity

  • Human

  • Mouse

With BSA and Azide
Conjugation

  • unconjugated

Overview

Product Name

FGFR2 Recombinant Rabbit Monoclonal Antibody [PSH26-90] - BSA and Azide free

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Recombinant protein within human FGFR2 aa 1-400.

Species Reactivity

Human, Mouse

Validated Applications

WB, FC, IP

Target Molecular Weight

Predicted band size: 92 kDa

Positive Control

Saos-2 (Human osteosarcoma cell) cell lysate, HT-29 (Human colorectal adenocarcinoma cell) cell lysate, HCT 116 (Human colon cancer cell) cell lysate, NIH/3T3 (Mouse fibroblast) cell lysate.

Conjugation

unconjugated

Clone Number

PSH26-90

Product Features

Form

Liquid

Concentration

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

Storage Instructions

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

Storage Buffer

PBS (pH7.4).

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:1,000-1:5,000

  • FC

  • 1:1,000

  • IP

  • 1-2μg/sample

Target

Function

Acidic and basic fibroblast growth factors (FGFs) are members of a family of multifunctional polypeptide growth factors that stimulate proliferation of cells of mesenchymal, epithelial and neuroectodermal origin. Like other growth factors, FGFs act by binding and activating specific cell surface receptors. These include the Flg receptor or FGFR-1, the Bek receptor (or FGFR-2), FGFR-3, FGFR-4, FGFR-5 and FGFR-6. These receptors usually contain an extracellular ligand-binding region containing three immunoglobulin-like domains, a transmembrane domain and a cytoplasmic tyrosine kinase domain. The gene encoding human Bek (also designated K-sam) maps to chromosome 10q26.13 and is alternatively spliced to produce several isoforms. Heterogeneous mutations in Bek are associated with a range of craniosynostosis syndromes including Pfeiffer syndrome, Crouzon syndrome, Jackson-Weiss syndrome and Apert syndrome..

Background References

1. Ramsey MR et al. FGFR2 signaling underlies p63 oncogenic function in squamous cell carcinoma. J Clin Invest 123:3525-38 (2013).

2. Huang Y et al. Twist1- and twist2-haploinsufficiency results in reduced bone formation. PLoS One 9:e99331 (2014).

Subcellular Location

Cell membrane, Golgi apparatus, Cytoplasmic vesicle.

UNIPROT

SwissProt: P21802 Human

SwissProt: P21803 Mouse

Synonyms

bacteria-expressed kinase antibody

BBDS antibody

BEK antibody

BEK fibroblast growth factor receptor antibody

BFR1 antibody

CD332 antibody

CD332 antigen antibody

CEK3 antibody

CFD1 antibody

Craniofacial dysostosis 1 antibody

Expand

Images

  • <span style="font-weight: bold;">☑ Relative expression (RE)</span><br /><br />Western blot analysis of FGFR2 on different lysates with Rabbit anti-FGFR2 antibody (<a href="/products/HA752052" style="font-weight: bold;text-decoration: underline;">HA752052</a>) at 1/1,000 dilution. <br /> <br />Lane 1: Saos-2 (Human osteosarcoma cell) cell lysate <br />Lane 2: SW620 (Human colon cancer cell) cell lysate <br />Lane 3: HT-29 (Human colorectal adenocarcinoma cell) cell lysate <br />Lane 4: HCT 116 (Human colon cancer cell) cell lysate <br />Lane 5: NIH/3T3 (Mouse fibroblast) cell lysate <br /> <br />Lysates/proteins at 15 µg/Lane. <br />Exposure time: 2 seconds; ECL: K1801 <br /> <br />FGFR2/KGFR/K-sam recognizes endogenous levels of total FGFR2 protein and a C-terminal truncated form K-sam (PMID: 17505008). <br />Negative control: SW620 (PMID: 24968263) <br /> <br />Blocking: 5% NFDM/TBST, 1 hour at room temperature <br />Primary antibody: <a href="/products/HA752052" style="font-weight: bold;text-decoration: underline;">HA752052</a>, 1/1,000 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: 92 kDa <br />Observed band size: 145 kDa

    ☑ Relative expression (RE)

    Western blot analysis of FGFR2 on different lysates with Rabbit anti-FGFR2 antibody (HA752052) at 1/1,000 dilution.

    Lane 1: Saos-2 (Human osteosarcoma cell) cell lysate
    Lane 2: SW620 (Human colon cancer cell) cell lysate
    Lane 3: HT-29 (Human colorectal adenocarcinoma cell) cell lysate
    Lane 4: HCT 116 (Human colon cancer cell) cell lysate
    Lane 5: NIH/3T3 (Mouse fibroblast) cell lysate

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

    FGFR2/KGFR/K-sam recognizes endogenous levels of total FGFR2 protein and a C-terminal truncated form K-sam (PMID: 17505008).
    Negative control: SW620 (PMID: 24968263)

    Blocking: 5% NFDM/TBST, 1 hour at room temperature
    Primary antibody: HA752052, 1/1,000 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: 92 kDa
    Observed band size: 145 kDa

  • <span style="font-weight: bold;">☑ Cell treatment (CT)</span><br /><br />Western blot analysis of FGFR2 on different lysates with Rabbit anti-FGFR2 antibody (<a href="/products/HA752052" style="font-weight: bold;text-decoration: underline;">HA752052</a>) at 1/5,000 dilution. <br /> <br />Lane 1: Saos-2 (Human osteosarcoma cell) cell lysate <br />Lane 2: Saos-2 treated with Deglycosylation cell lysate <br /> <br />Lysates/proteins at 15 µg/Lane. <br />Exposure time: 1 minute ; ECL: K1802 <br /> <br />Blocking: 5% NFDM/TBST, 1 hour at room temperature <br />Primary antibody: <a href="/products/HA752052" style="font-weight: bold;text-decoration: underline;">HA752052</a>, 1/5,000 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: 92 kDa <br />Observed band size: 145 kDa

    ☑ Cell treatment (CT)

    Western blot analysis of FGFR2 on different lysates with Rabbit anti-FGFR2 antibody (HA752052) at 1/5,000 dilution.

    Lane 1: Saos-2 (Human osteosarcoma cell) cell lysate
    Lane 2: Saos-2 treated with Deglycosylation cell lysate

    Lysates/proteins at 15 µg/Lane.
    Exposure time: 1 minute ; ECL: K1802

    Blocking: 5% NFDM/TBST, 1 hour at room temperature
    Primary antibody: HA752052, 1/5,000 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: 92 kDa
    Observed band size: 145 kDa

  • <span style="font-weight: bold;">☑ Relative expression (RE)</span><br /><br />Application: Flow Cytometry<br /><br />Species: Human<br />Sample: SW620 (Human colon cancer cell) (Left, Negative) / Saos-2 (Human osteosarcoma cell) (Right)<br /><br />Blocking: 1% BSA + 10% normal goat serum + human FcR (Miltenyi Biotec), 15 minutes at room temperature. <br />Antibody dilution buffer: 1x PBS.<br /><br />Primary antibody: <a href="/products/HA752052" style="font-weight: bold;text-decoration: underline;">HA752052</a> (1/1,000, 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.<br /><br />Negative control: SW620 (PMID: 24968263).

    ☑ Relative expression (RE)

    Application: Flow Cytometry

    Species: Human
    Sample: SW620 (Human colon cancer cell) (Left, Negative) / Saos-2 (Human osteosarcoma cell) (Right)

    Blocking: 1% BSA + 10% normal goat serum + human FcR (Miltenyi Biotec), 15 minutes at room temperature.
    Antibody dilution buffer: 1x PBS.

    Primary antibody: HA752052 (1/1,000, 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.

    Negative control: SW620 (PMID: 24968263).

  • Immunoprecipitation (IP)<br /><br />FGFR2 was immunoprecipitated in 0.2 mg Saos-2 (Human osteosarcoma cell) cell lysate with <a href="/products/HA752052" style="font-weight: bold;text-decoration: underline;">HA752052</a> at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using <a href="/products/HA752052" style="font-weight: bold;text-decoration: underline;">HA752052</a> at 1/5,000 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: Saos-2 cell lysate (input)<br />Lane 2: <a href="/products/HA752052" style="font-weight: bold;text-decoration: underline;">HA752052</a> IP in Saos-2 cell lysate<br />Lane 3: Rabbit IgG instead of <a href="/products/HA752052" style="font-weight: bold;text-decoration: underline;">HA752052</a> in Saos-2 cell lysate<br /><br />Exposure time: 3 minutes<br />Blocking: 5% NFDM/TBST, 1 hour at room temperature<br /><br />Primary dilution: <a href="/products/HA752052" style="font-weight: bold;text-decoration: underline;">HA752052</a>, 1/5,000 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: 92 kDa<br />Observed band size: 145 kDa

    Immunoprecipitation (IP)

    FGFR2 was immunoprecipitated in 0.2 mg Saos-2 (Human osteosarcoma cell) cell lysate with HA752052 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA752052 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: Saos-2 cell lysate (input)
    Lane 2: HA752052 IP in Saos-2 cell lysate
    Lane 3: Rabbit IgG instead of HA752052 in Saos-2 cell lysate

    Exposure time: 3 minutes
    Blocking: 5% NFDM/TBST, 1 hour at room temperature

    Primary dilution: HA752052, 1/5,000 in primary antibody dilution buffer (K1803), 2 hours at room temperature

    Predicted band size: 92 kDa
    Observed band size: 145 kDa

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