Anti-FGF23 antibody [EPR25309-57] - BSA and Azide free

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-FGF23 antibody [EPR25309-57] - BSA and Azide free (AB307421)

This data was developed using ab307420, the same antibody clone in a different buffer formulation. Immunofluorescent analysis of 100% methanol-fixed, 0.1% TritonX-100 permeABilized HaCaT (human skin keratinocyte) cells lABelling FGF 23 with ab307420 at 1/50 (9.96 ug/ml) dilution, followed by ab150081 Goat Anti-RABbit IgG H&L (Alexa Fluor? 488) preadsorbed antibody at 1/1000 2 ug/ml dilution (Green). Confocal image showing membranous and weak cytoplasmic staining in HaCaT cells.The image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). is observed. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor? 594) was used to counterstain tubulin at 1/200 2.5 ug/ml dilution (Red). The Nuclear counterstain was DAPI (Blue). Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-RABbit IgG H&L (Alexa Fluor? 488) preadsorbed at 1/1000 2 ug/ml dilution.

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-FGF23 antibody [EPR25309-57] - BSA and Azide free (AB307421)

This data was developed using ab307420, the same antibody clone in a different buffer formulation. Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HaCaT (human skin keratinocyte) cells labeling FGF 23 with ab307420 at 1/500 dilution (0.1 µg)  (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/2000 dilution was used as the secondary antibody.

• WB

Supplier Data

Western blot - Anti-FGF23 antibody [EPR25309-57] - BSA and Azide free (AB307421)

This data was developed using ab307420, the same antibody clone in a different buffer formulation.

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

This blot was developed using a high sensitivity ECL substrate. The high-sensitivity ECL substrate used allows for the detection of proteins in the mid-femtogram range.

The identity of the bands between 90 kDa and 160 kDa are unknown.

All lanes:

Western blot - Anti-FGF23 antibody [EPR25309-57] (<a href='/en-us/products/primary-antibodies/fgf23-antibody-epr25309-57-ab307420'>ab307420</a>) at 1/1000 dilution

Lane 1:

HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

HaCaT (human skin keratinocyte) whole cell lysate at 20 µg

Lane 3:

HT-29 (human colorectal adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 4:

PC-3 (human prostate adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 5:

RAW264.7 (mouse elson murine leukemia virus-induced tumor macrophage) whole cell lysate at 20 µg

Lane 6:

bEnd.3 (mouse brain endothelioma) whole cell 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 1/100000 dilution

Observed band size: 33 kDa

true

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-FGF23 antibody [EPR25309-57] - BSA and Azide free (AB307421)

This data was developed using ab307420, the same antibody clone in a different buffer formulation.

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

This blot was developed using a high sensitivity ECL substrate. The high-sensitivity ECL substrate used allows for the detection of proteins in the mid-femtogram range.

The identity of the higher MW band at approximately 140 kDa may represent FGF23 oligermisation. This has not been confirmed with additional tests.

All lanes:

Western blot - Anti-FGF23 antibody [EPR25309-57] (<a href='/en-us/products/primary-antibodies/fgf23-antibody-epr25309-57-ab307420'>ab307420</a>) at 1/1000 dilution

Lane 1:

HeLa (human cervix adenocarcinoma epithelial cell) transfected with scrambled siRNA control whole cell lysate at 20 µg

Lane 2:

HeLa transfected with siRNA specifically targeti FGF23 whole cell 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 1/100000 dilution

Observed band size: 33 kDa

true

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-FGF23 antibody [EPR25309-57] - BSA and Azide free (AB307421)

This data was developed using 307420, the same antibody clone in a different buffer formulation.

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

Negative control : brain (PMID : 20667984).

This blot was developed using a high sensitivity ECL substrate. The high-sensitivity ECL substrate used allows for the detection of proteins in the mid-femtogram range.

All lanes:

Western blot - Anti-FGF23 antibody [EPR25309-57] (<a href='/en-us/products/primary-antibodies/fgf23-antibody-epr25309-57-ab307420'>ab307420</a>) at 1/1000 dilution

Lane 1:

Mouse thymus tissue lysate at 20 µg

Lane 2:

Mouse brain 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 1/100000 dilution

Observed band size: 33 kDa

true

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-FGF23 antibody [EPR25309-57] - BSA and Azide free (AB307421)

This data was developed using 307420, the same antibody clone in a different buffer formulation. Blocking and diluting buffer and concentration : 10 seconds Exposure time :

All lanes:

Western blot - Anti-FGF23 antibody [EPR25309-57] (<a href='/en-us/products/primary-antibodies/fgf23-antibody-epr25309-57-ab307420'>ab307420</a>) at 1/1000 dilution

All lanes:

293T cells transfected with a human FGF23 expression vector containi a myc-his tag whole cell lysate 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 1/100000 dilution

Observed band size: 33 kDa

false

Exposure time: 10s