Anti-GDF15 antibody [EPR19939]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GDF15 antibody [EPR19939] (AB206414)

Immunohistochemical analysis of paraffin-embedded human prostate hyperplasia tissue labeling GDF15 with ab206414 at 1/100 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Cytoplasmic staining on human prostate hyperplasia is observed (PMID : 9593718). Positive staining on human placenta and prostate hyperplasia other tissues tested were negative.

Counter stained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GDF15 antibody [EPR19939] (AB206414)

Immunohistochemical analysis of paraffin-embedded human placenta tissue labeling GDF15 with ab206414 at 1/100 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Cytoplasmic staining on human placenta is observed (PMID : 9593718). Positive staining on human placenta and prostate hyperplasia other tissues tested were negative.

Counter stained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GDF15 antibody [EPR19939] (AB206414)

Immunohistochemical analysis of formalin fixed paraffin embedded human prostate labelling GDF15 with ab206414 at a concentration of 3µg/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. ab206414 anti-GDF15 antibody [EPR19939] 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).

• IP

Supplier Data

Immunoprecipitation - Anti-GDF15 antibody [EPR19939] (AB206414)

GDF15 was immunoprecipitated from 0.35 mg of HepG2 (Human liver hepatocellular carcinoma cell line) whole cell lysate with ab206414 at 1/30 dilution.

Western blot was performed from the immunoprecipitate using ab206414 at 1/1000 dilution.

VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1000 dilution.

Lane 1 : HepG2 whole cell lysate 10μg (Input).

Lane 2 : ab206414 IP in HepG2 whole cell lysate.

Lane 3 : Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) instead of ab206414 in HepG2 whole cell lysate.

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

Exposure time : 10 seconds.

All lanes:

Immunoprecipitation - Anti-GDF15 antibody [EPR19939] (ab206414)

Predicted band size: 34 kDa

false

• WB

Supplier Data

Western blot - Anti-GDF15 antibody [EPR19939] (AB206414)

Blocking/Dilution buffer : 5% NFDM/TBST.

Exposure time : Lane 1 : 10 seconds; Lane 2 : 3 minutes.

The molecular weight observed is consistent with the literature (PMID : 20884666).

All lanes:

Western blot - Anti-GDF15 antibody [EPR19939] (ab206414) at 1/1000 dilution

Lane 1:

Human prostate cancer lysate at 10 µg

Lane 2:

Human placenta lysate at 10 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/10000 dilution

Predicted band size: 34 kDa

Observed band size: 12.5 kDa

false

• WB

Unknown

Western blot - Anti-GDF15 antibody [EPR19939] (AB206414)

Blocking/Dilution buffer : 5% NFDM/TBST.

The expression level of GDF15 can be increased by TPA treatment (PMID : 15757899).

All lanes:

Western blot - Anti-GDF15 antibody [EPR19939] (ab206414) at 1/1000 dilution

Lane 1:

Untreated HT1080 (Human fibrosarcoma cell line) whole cell lysate at 10 µg

Lane 2:

HT1080 (Human fibrosarcoma cell line) whole cell lysate treated with 200ng/ml Phorbol-12-myristate-13-acetate (TPA) for 24 hours at 10 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 1/100000 dilution

Predicted band size: 34 kDa

Observed band size: 35 kDa

false

Exposure time: 3s

• WB

Supplier Data

Western blot - Anti-GDF15 antibody [EPR19939] (AB206414)

Blocking/Dilution buffer : 5% NFDM/TBST.

Exposure time : Lane 1 : 3 minutes; Lane 2 : 10 seconds.

The molecular weight observed is consistent with the literature (PMID : 20884666).

All lanes:

Western blot - Anti-GDF15 antibody [EPR19939] (ab206414) at 1/1000 dilution

Lane 1:

HepG2 (Human liver hepatocellular carcinoma cell line) whole cell lysate at 20 µg

Lane 2:

LNCaP (Human prostate cancer cell line) whole cell lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/10000 dilution

Predicted band size: 34 kDa

Observed band size: 12.5 kDa,35 kDa

false

• WB

Supplier Data

Western blot - Anti-GDF15 antibody [EPR19939] (AB206414)

Blocking/Dilution buffer : 5% NFDM/TBST.

Both pro and mature form can be increased by TPA treatment.

All lanes:

Western blot - Anti-GDF15 antibody [EPR19939] (ab206414) at 1/1000 dilution

Lane 1:

Untreated HT1080 (Human fibrosarcoma cell line) whole cell lysate at 10 µg

Lane 2:

HT1080 (Human fibrosarcoma cell line) whole cell lysate treated with 200ng/ml Phorbol-12-myristate-13-acetate (TPA) for 24 hours at 10 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 1/100000 dilution

Predicted band size: 34 kDa

Observed band size: 35 kDa

false

Exposure time: 1min

• ELISA

Lab

ELISA - Anti-GDF15 antibody [EPR19939] (AB206414)

ELISA analysis of ab206414 binding functionally active mature GDF15 (ab50077) at 1 μg/mL.