Anti-IGFBP6 antibody [EPR21809]

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-IGFBP6 antibody [EPR21809] (AB219560)

Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol-permeabilized HaCaT (human keratinocyte cell line) cell line labeling IGFBP6 with ab219560 at 1/500 dilution (red) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabeled control (cells incubated with secondary antibody only) (blue). Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) at 1/2000 dilution was used as the secondary antibody.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-IGFBP6 antibody [EPR21809] (AB219560)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HaCaT (human keratinocyte cell line) cells labeling IGFBP6 with ab219560 at 1/100 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic and weakly nuclear staining in HaCaT cell line.

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

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.

• WB

Supplier Data

Western blot - Anti-IGFBP6 antibody [EPR21809] (AB219560)

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

The molecular mass observed is consistent with the literature (PMID 27623076; PMID 21820463).

All lanes:

Western blot - Anti-IGFBP6 antibody [EPR21809] (ab219560) at 1/1000 dilution

Lane 1:

Human placenta lysate at 20 µg

Lane 2:

Human kidney lysate at 20 µg

Lane 3:

NCI-H1299 (human lung carcinoma cell line) whole cell lysate at 20 µg

Lane 4:

HaCaT (human keratinocyte cell line) 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

Predicted band size: 25 kDa

Observed band size: 34 kDa

true

Exposure time: 59s

• WB

Supplier Data

Western blot - Anti-IGFBP6 antibody [EPR21809] (AB219560)

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

All lanes:

Western blot - Anti-IGFBP6 antibody [EPR21809] (ab219560) at 1/5000 dilution

All lanes:

Human IGFBP6 recombinant protein (aa25-240) from mammalian expression system at 0.01 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: 25 kDa

Observed band size: 34 kDa

true

Exposure time: 15s