Anti-ApoER2 antibody [EPR3326] - BSA and Azide free

• IP

Unknown

Immunoprecipitation - Anti-ApoER2 antibody [EPR3326] - BSA and Azide free (AB215274)

ab108208 (purified) at 1/40 immunoprecipitating ApoER2 in SH-SY5Y (human neuroblastoma) whole cell lysate.

Lane 1 (input) : SH-SY5Y whole cell lysate 10ug

Lane 2 (+) : ab108208 + SH-SY5Y whole cell lysate

Lane 3 (-) : Rabbit monoclonal IgG (ab172730) instead of ab108208 in SH-SY5Y whole cell lysate

For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10,000 dilution.

Blocking buffer and concentration : 5% NFDM/TBST.

Diluting buffer and concentration : 5% NFDM /TBST.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab108208).

All lanes:

Immunoprecipitation - Anti-ApoER2 antibody [EPR3326] (<a href='/en-us/products/primary-antibodies/apoer2-antibody-epr3326-ab108208'>ab108208</a>)

Predicted band size: 106 kDa

Observed band size: 130-180 kDa

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• WB

Lab

Western blot - Anti-ApoER2 antibody [EPR3326] - BSA and Azide free (AB215274)

Lanes 1 - 2 : Merged signal (red and green). Green - ab108208 observed at 106 kDa. Red - loading control, ab9484, observed at 37 kDa.

ab108208 was shown to specifically react with ApoER2 in wild-type HAP1 cells as signal was lost in LRP8 knockout cells. Wild-type and ApoER2 knockout samples were subjected to SDS-PAGE. ab108208 and ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab108208).

All lanes:

Western blot - Anti-ApoER2 antibody [EPR3326] - BSA and Azide free (ab215274) at 1/1000 dilution

Lane 1:

Wild-type HAP1 whole cell lysate at 40 µg

Lane 2:

ApoER2 knockout HAP1 whole cell lysate at 40 µg

Predicted band size: 106 kDa

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• WB

Unknown

Western blot - Anti-ApoER2 antibody [EPR3326] - BSA and Azide free (AB215274)

Blocking and diluting buffer 5% NFDM/TBST.

130kDa : Immature form;

170kDa : Glycosylated mature form. (PMID : 24344333, 24532792, 24705369 and 25233900)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab108208).

All lanes:

Western blot - Anti-ApoER2 antibody [EPR3326] (<a href='/en-us/products/primary-antibodies/apoer2-antibody-epr3326-ab108208'>ab108208</a>) at 1/10000 dilution

Lane 1:

SH-SY5Y (human neuroblastoma) whole cell lysate at 20 µg

Lane 2:

Human fetal brain tissue lysate at 20 µg

Lane 3:

Mouse brain tissue lysate at 20 µg

Lane 4:

C6 (rat glioma) whole cell lysate at 20 µg

Lane 5:

Neuro-2a (mouse neuroblastoma) whole cell lysate at 20 µg

Secondary

All lanes:

Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/2000 dilution

Predicted band size: 106 kDa

Observed band size: 130-170 kDa

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