Amphiregulin Recombinant Rabbit Monoclonal Antibody [PSH19-73]
cat.: HA751728
| Product Type: | Recombinant Rabbit monoclonal IgG, primary antibodies |
|---|---|
| Species reactivity: | Human, Mouse, Rat |
| Applications: | WB, IF-Cell, FC, IP |
| Clonality: | Monoclonal |
| Clone number: | PSH19-73 |
| Form: | Liquid |
| Storage condition: | Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles. |
| Storage buffer: | 1*PBS (pH7.4). |
| Concentration: | 1ug/ul |
| Purification: | Protein A affinity purified. |
| Molecular weight: | Predicted band size: 28 kDa |
| Isotype: | IgG |
| Immunogen: | Recombinant protein within human Amphiregulin aa 1-187. |
| Positive control: | HCT 116 treated with 100ng/mL TPA for 48 hours add 300ng/mL BFA for last 24 hours cell lysate, A549 treated with 500ng/mL Ionomycin Ca2+ Salt and 50ng/mL TPA for 6 hours add 500ng/mL BFA for last 3 hours cell lysate, C2C12 cell lysate, C2C12 treated with 100ng/mL TPA for 48 hours add 300ng/mL BFA for last 24 hours cell lysate, EL4 cell lysate, EL4 treated with treated with 500ng/mL Ionomycin Ca2+ Salt and 50ng/mL TPA for 24 hours add 500ng/mL BFA for last 20 hours cell lysate, Mouse brain tissue lysate. |
| Subcellular location: | Membrane. |
| Recommended Dilutions:
WB IF-Cell FC |
1:5,000 1:2,000-1:5,000 1:5,000 |
| Uniprot #: | SwissProt: P15514 Human | P31955 Mouse | P24338 Rat |
| Alternative names: | A REG Amphiregulin Amphiregulin B AR AREG AREG_HUMAN AREGB Colorectum cell derived growth factor Colorectum cell-derived growth factor CRDGF MGC13647 Schwannoma derived growth factor SDGF |
Images
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Fig1:
Western blot analysis of Amphiregulin on different lysates with Rabbit anti-Amphiregulin antibody (HA751728) at 1/5,000 dilution. Lane 1: HCT 116 cell lysate Lane 2: HCT 116 treated with 100ng/mL TPA for 48 hours add 300ng/mL BFA for last 24 hours cell lysate Lane 3: A549 cell lysate Lane 4: A549 treated with 500ng/mL Ionomycin Ca2+ Salt and 50ng/mL TPA for 6 hours add 500ng/mL BFA for last 3 hours cell lysate Lane 5: C2C12 cell lysate Lane 6: C2C12 treated with 100ng/mL TPA for 48 hours add 300ng/mL BFA for last 24 hours cell lysate Lane 7: EL4 cell lysate Lane 8: EL4 treated with treated with 500ng/mL Ionomycin Ca2+ Salt and 50ng/mL TPA for 24 hours add 500ng/mL BFA for last 20 hours cell lysate Lane 9: Mouse brain tissue lysate Lysates/proteins at 20 µg/Lane. Predicted band size: 28 kDa Observed band size: 40 kDa Exposure time: 10 seconds; ECL: K1801; 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 (HA751728) at 1/5,000 dilution was used in primary antibody dilution (K1803) at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. |
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Fig2:
Flow cytometric analysis of HCT 116 cells untreated (left) / treated with 100ng/mL TPA for 48 hours add 300ng/mL BFA for last 24 hours (right) labeling Amphiregulin. Cells were fixed and permeabilized. Then stained with the primary antibody (HA751728, 1/5,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black). |
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Fig3:
Immunocytochemistry analysis of HCT 116 cells untreated / treated with 100ng/mL TPA for 48 hours add 300ng/mL BFA for last 24 hours labeling Amphiregulin with Rabbit anti-Amphiregulin antibody (HA751728) at 1/5,000 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Amphiregulin antibody (HA751728) at 1/5,000 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution. |
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Fig4:
Immunocytochemistry analysis of NR8383 cells untreated / treated with 100nM TPA overnight then treated with 100ng/mL LPS for 7 hours add 1μg/mL BFA for last 3 hours labeling Amphiregulin with Rabbit anti-Amphiregulin antibody (HA751728) at 1/5,000 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Amphiregulin antibody (HA751728) at 1/5,000 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution. |
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Fig5:
Amphiregulin was immunoprecipitated from 0.2 mg HCT 116 treated with 100ng/mL TPA for 48 hours add 300ng/mL BFA for last 24 hours cell lysate with HA751728 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA751728 at 1/5,000 dilution. HRP Conjugated Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature. Lane 1: HCT 116 treated with 100ng/mL TPA for 48 hours add 300ng/mL BFA for last 24 hours cell lysate (input) Lane 2: HA751728 IP in HCT 116 treated with 100ng/mL TPA for 48 hours add 300ng/mL BFA for last 24 hours cell lysate Lane 3: Rabbit IgG instead of HA751728 in HCT 116 treated with 100ng/mL TPA for 48 hours add 300ng/mL BFA for last 24 hours cell lysate Blocking/Dilution buffer: primary antibody dilution (K1803) Exposure time: 6 seconds; ECL: K1801 |
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.