Anti-RBP4 antibody [EPR18020-115]

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RBP4 antibody [EPR18020-115] (AB188230)

Immunohistochemical analysis of paraffin-embedded (A) Kidney tissue from wild-type C57BL/6JGpt mice and (B) Kidney tissue from Rbp4 knockout mice tissue labeling RBP4 with ab188230 at 1/50 dilution, followed by a ready to use LeicaDS9800 (Bond^™ Polymer Refine Detection).

Positive staining on (A) Kidney tissue from wild-type C57BL/6JGpt mice and no staining on (B) Kidney tissue from Rbp4 knockout mice.
The section was incubated with ab188230 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND^® RX instrument.
The tissue samples were kindly provided by GemPharmatech. C57BL/6JGpt wildtype mice and Rbp4-KO homozygous mice (Strain ID : T012705).

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond^™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RBP4 antibody [EPR18020-115] (AB188230)

Immunohistochemical analysis of paraffin-embedded mouse liver tissue labeling RBP4 with ab188230 at 1/50 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Cytoplasmic staining on hepatocytes of mouse liver (PMID : 19624771; PMID : 21324757) is observed. Counter stained with hematoxylin.

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

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

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-RBP4 antibody [EPR18020-115] (AB188230)

Intracellular flow cytometric analysis of4% paraformaldehyde-fixed, 90% methanol-permeabilized Hepa1-6 (mouse hepatoma epithelial cell line) cell line labeling RBP4 with ab188230 at 1/500 dilution (red) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat Anti-Rabbit IgG H&L (Alexa Fluor^®488) (ab150077) at 1/2000 dilution was used as the secondary antibody.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RBP4 antibody [EPR18020-115] (AB188230)

Immunohistochemical analysis of paraffin-embedded rat liver tissue labeling RBP4 with ab188230 at 1/50 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Cytoplasmic staining on hepatocytes of rat liver (PMID : 19624771; PMID : 21324757) is observed. Counter stained with hematoxylin.

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

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

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-RBP4 antibody [EPR18020-115] (AB188230)

Immunofluorescent analysis of 4% paraformalehyde-fixed, 0.1% Triton X-100 permeabilized Hepa1-6 (mouse hepatoma epithelial cell line) cells labeling RBP4 with ab188230 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining on Hepa1-6 cells.

The nuclear counterstain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) (ab195889) at 1/200 dilution (red).

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution.

• IP

Supplier Data

Immunoprecipitation - Anti-RBP4 antibody [EPR18020-115] (AB188230)

RBP4 was immunoprecipitated from 0.35 mg of Hepa1-6 (mouse hepatoma epithelial cell line) whole cell lysate with ab188230 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab188230 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1000 dilution.

Lane 1 : Hepa1-6 whole cell lysate 10 μg (Input).

Lane 2 : ab188230 IP in Hepa1-6 whole cell lysate.

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab188230 in Hepa1-6 whole cell lysate.

Exposure time : 2 seconds.

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

All lanes:

Immunoprecipitation - Anti-RBP4 antibody [EPR18020-115] (ab188230)

Predicted band size: 23 kDa

Observed band size: 21 kDa

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• WB

Lab

Western blot - Anti-RBP4 antibody [EPR18020-115] (AB188230)

The tissue samples were kindly provided by GemPharmatech. C57BL/6JGpt wildtype mice and Nppa-KO homozygous mice (Strain ID : T012705).

The identity of the higher MW band at approximately 37 kDa (in Lane 4-5) is unknown.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

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

All lanes:

Western blot - Anti-RBP4 antibody [EPR18020-115] (ab188230) at 1/1000 dilution

Lane 1:

Wild-type Mouse kidney (female)

Lane 2:

Rbp4 knockout kidneytissue lysate (female case1)

Lane 3:

Rbp4 knockout kidneytissue lysate (female case2)

Lane 4:

Wild-type Mouse liver (female)

Lane 5:

Rbp4 knockout liver tissue lysate (female case1)

Lane 6:

Rbp4 knockout liver tissue lysate (female case2)

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 1/5000 dilution

Observed band size: 21 kDa

false

Exposure time: 92s

• WB

Supplier Data

Western blot - Anti-RBP4 antibody [EPR18020-115] (AB188230)

Exposure times : Lanes 1, 2 and 4 : 1 minute; Lane 3 : 30 seconds.

Blocking/dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-RBP4 antibody [EPR18020-115] (ab188230) at 1/1000 dilution

Lane 1:

Mouse serum at 20 µg

Lane 2:

Mouse kidney tissue lysate at 20 µg

Lane 3:

Hepa1-6 (mouse hepatoma epithelial cell line) whole cell lysate at 20 µg

Lane 4:

Rat kidney 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

Predicted band size: 23 kDa

Observed band size: 21 kDa

true