Anti-ABCA1 antibody [EPR27494-57] - BSA and Azide free

• WB

Lab

Western blot - Anti-ABCA1 antibody [EPR27494-57] - BSA and Azide free (AB307537)

This data was developed using ab307536, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer : 5% NFDM/TBST.

Exposure : lanes 1-4 : 180 seconds; lanes : 5-8; 10 seconds

The higher band is probably glycosylated form (PMID : 16873719).

Compared with ab307536, ab307534 has higher sensitivity.

We recommend ab307534 as an alternative in testing human lysate, or optimize experimental protocols (increasing lysate amount, using lower dilution or higher sensitivity ECL substrate) to get visible band.

Lanes 1 - 4:

Western blot - Anti-ABCA1 antibody [EPR27494-57] (<a href='/en-us/products/primary-antibodies/abca1-antibody-epr27494-57-ab307536'>ab307536</a>) at 1/1000 dilution

Lanes 5 - 8:

Western blot - Anti-ABCA1 antibody [EPR27494-51] (<a href='/en-us/products/primary-antibodies/abca1-antibody-epr27494-51-ab307534'>ab307534</a>) at 1/1000 dilution

Lane 6:

THP-1 treated first with 100ng/ml PAM for 72 hours and then replaced with 100ng/ml LPS for 5 hours, 300ng/ml BFA was then added for additional 3 hours whole cell lysate at 20 µg

Lane 7:

Untreated HepG2 (Human hepatocellular carcinoma epithelial cell) whole cell lysate at 20 µg

Lanes 1 and 5:

Untreated THP-1 (Human monocytic leukemia monocyte) whole cell lysate at 20 µg

Lane 2:

THP-1 treated first with 100ng/ml PAM for 72 hours and then replaced with 100ng/ml LPS for 5 hours, 300ng/ml BFA was then added for additional 3 hours whole cell lysate at 20 µg

Lane 3:

Untreated HepG2 (Human hepatocellular carcinoma epithelial cell) whole cell lysate at 20 µg

Lanes 4 and 8:

HepG2 treated with 20uM T0901317 for 16 hours 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 1/20000 dilution

Predicted band size: 254 kDa

Observed band size: 254 kDa,400 kDa

false

• WB

Supplier Data

Western blot - Anti-ABCA1 antibody [EPR27494-57] - BSA and Azide free (AB307537)

This data was developed using 307536, the same antibody clone in a different buffer formulation. Blocking and diluting buffer and concentration : Negative control : 293T(PMID : 11896206), SH-SY5Y(PMID : 23600914) Bands around 500 kDa may due to aggregation (PMID : 31641056, PMID : 29103206). Bands below 260 kDa may due to degradation. Samples are non-boiled as boiling may cause protein aggregates. 48 seconds Exposure time :

All lanes:

Western blot - Anti-ABCA1 antibody [EPR27494-57] (<a href='/en-us/products/primary-antibodies/abca1-antibody-epr27494-57-ab307536'>ab307536</a>) at 1/1000 dilution

Lane 1:

HepG2 (human hepatocellar carcinoma epithelial cell) whole cell lysate 40 μg

Lane 2:

U-87 MG (human glioblastoma-astrocytoma epithelial cell) whole cell lysate 40 μg

Lane 3:

293T (human embryonic kidney epithelial cell) whole cell lysate 40 μg

Lane 4:

SH-SY5Y (human neuroblastoma epithelial cell) whole cell lysate 40 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: 260 kDa

false

Exposure time: 48s

• WB

Supplier Data

Western blot - Anti-ABCA1 antibody [EPR27494-57] - BSA and Azide free (AB307537)

This data was developed using 307536, the same antibody clone in a different buffer formulation. Blocking and diluting buffer and concentration : ABCA1 expression was increased after treatment with the nonsteroidal LXR agonist T0901317 (PMID : 12384498). Bands around 500 kDa may due to aggregation (PMID : 31641056, PMID : 29103206). Samples are non-boiled as boiling may cause protein aggregates. Lanes 1 : 37 seconds, Lane 2 and 3 : 103 seconds. Exposure time :

All lanes:

Western blot - Anti-ABCA1 antibody [EPR27494-57] (<a href='/en-us/products/primary-antibodies/abca1-antibody-epr27494-57-ab307536'>ab307536</a>) at 1/1000 dilution

Lane 1:

Mouse liver tissue lysate 50 μg

Lane 2:

Untreated Neuro-2a (mouse neuroblastoma neuroblast) whole cell lysate 40 μg

Lane 3:

Neuro-2a treated with 1uM T0901317 for 24 hours whole cell lysate 40 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: 260 kDa

false

Exposure time: 37s

• WB

Supplier Data

Western blot - Anti-ABCA1 antibody [EPR27494-57] - BSA and Azide free (AB307537)

This data was developed using 307536, the same antibody clone in a different buffer formulation. Blocking and diluting buffer and concentration : This antibody does not cross-react with human ABCA3, ABCA4 and ABCA7. Samples are non-boiled as boiling may cause protein aggregates. 15 seconds Exposure time :

All lanes:

Western blot - Anti-ABCA1 antibody [EPR27494-57] (<a href='/en-us/products/primary-antibodies/abca1-antibody-epr27494-57-ab307536'>ab307536</a>) at 1/1000 dilution

Lane 1:

293T (human embryonic kidney epithelial cell) whole cell lysate 20 μg

Lane 2:

293T cells transfected with an empty vector containi a myc-his tag whole cell lysate 20 μg

Lane 3:

293T cells transfected with a human CA1 expression vector containi a myc-his tag whole cell lysate 10 μg

Lane 4:

293T cells transfected with a human CA3 expression vector containi a myc-his tag whole cell lysate 36 μg

Lane 5:

293T cells transfected with a human CA4 expression vector containi a myc-his tag whole cell lysate 4 μg

Lane 6:

293T cells transfected with a human CA7 expression vector containi a myc-his tag whole cell lysate 4 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: 260 kDa

false

Exposure time: 15s