Anti-VLDL Receptor/VLDL-R antibody [EPR26178-72] (BSA and Azide free)

• IHC

Supplier Data

Immunohistochemistry - Anti-VLDL Receptor/VLDL-R antibody [EPR26178-72] (BSA and Azide free) (AB302918)

This data was developed using ab302917, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded Human liver tissue labeling VLDL Receptor/VLDL-R with ab302917 at 1/500 (0.978 ug/ml) followed by a LeicaDS9800 (Bond™ Polymer Refine Detection) was used at Ready to use dilution. Negative control : No staining on human liver. The section was incubated with ab302917 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is LeicaDS9800 (Bond™ Polymer Refine Detection) was used at Ready to use dilution. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• IHC

Supplier Data

Immunohistochemistry - Anti-VLDL Receptor/VLDL-R antibody [EPR26178-72] (BSA and Azide free) (AB302918)

This data was developed using ab302917, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human cardiac muscle tissue labeling VLDL Receptor/VLDL-R with ab302917 at 1/500 (0.978 ug/ml) followed by a LeicaDS9800 (Bond™ Polymer Refine Detection) was used at Ready to use dilution. Cytoplasmic staining on cardiac muscle. The section was incubated with ab302917 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is LeicaDS9800 (Bond™ Polymer Refine Detection) was used at Ready to use dilution. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• IHC

Supplier Data

Immunohistochemistry - Anti-VLDL Receptor/VLDL-R antibody [EPR26178-72] (BSA and Azide free) (AB302918)

This data was developed using ab302917, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded Human pancreas tissue labeling VLDL Receptor/VLDL-R with ab302917 at 1/500 (0.978 ug/ml) followed by a LeicaDS9800 (Bond™ Polymer Refine Detection) was used at Ready to use dilution. Cytoplasmic staining on islet of human pancreas. The section was incubated with ab302917 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is LeicaDS9800 (Bond™ Polymer Refine Detection) was used at Ready to use dilution. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• WB

Lab

Western blot - Anti-VLDL Receptor/VLDL-R antibody [EPR26178-72] (BSA and Azide free) (AB302918)

This data was developed using ab302917, the same antibody clone in a different buffer formulation.

Western blot : Rabbit Monoclonal [EPR26178-72] to VLDL Receptor/VLDL-R ab302917 staining at 1/1000 dilution, shown in green; Mouse anti GAPDH (ab8245) loading control staining at 1/20,000 dilution, shown in magenta.

A band was observed at 115/140 kDa in Wild-type U-87 MG UNBOILED cell lysates with no signal observed at this size in VLDLR knockout U-87 MG UNBOILED cell line.

To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3pc Milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged.

Secondary antibodies used were Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-VLDL Receptor/VLDL-R antibody [EPR26178-72] (<a href='/en-us/products/primary-antibodies/vldl-receptor-vldl-r-antibody-epr26178-72-ab302917'>ab302917</a>) at 1/1000 dilution

Lane 1:

Wild-type U-87 MG UNBOILED at 20 µg

Lane 2:

VLDLR knockout U-87 MG UNBOILED at 20 µg

Lane 2:

Western blot - Human VLDLR knockout U-87 MG cell line (<a href='/en-us/products/cell-lines/human-vldlr-knockout-u-87-mg-cell-line-ab306807'>ab306807</a>) at 20 µg

Lane 3:

THP-1 UNBOILED at 20 µg

Lane 4:

Daudi UNBOILED at 20 µg

Secondary

All lanes:

Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 96 kDa

Observed band size: 115 kDa,140 kDa

false

• WB

Lab

Western blot - Anti-VLDL Receptor/VLDL-R antibody [EPR26178-72] (BSA and Azide free) (AB302918)

This data was developed using ab302917, 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.

All lanes:

Western blot - Anti-VLDL Receptor/VLDL-R antibody [EPR26178-72] (<a href='/en-us/products/primary-antibodies/vldl-receptor-vldl-r-antibody-epr26178-72-ab302917'>ab302917</a>) at 1/1000 dilution

Lane 1:

Human VLDLR recombinant protein at 10 ng

Lane 2:

His-tagged human LDLR recombinant protein  at 10 ng

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: 96 kDa

Observed band size: 15-50 kDa

true

• WB

Lab

Western blot - Anti-VLDL Receptor/VLDL-R antibody [EPR26178-72] (BSA and Azide free) (AB302918)

This data was developed using ab302917, the same antibody clone in a different buffer formulation.

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

The expression of VLDLR is upregulated in response to Dexamethasone treatment (PMID : 11960750).

Low expression : Huh7 (PMID : 29289645)

Exposure time :

Lane 1 and 2 : 180 seconds

Lane 3 and 4 : 48 seconds

All lanes:

Western blot - Anti-VLDL Receptor/VLDL-R antibody [EPR26178-72] (<a href='/en-us/products/primary-antibodies/vldl-receptor-vldl-r-antibody-epr26178-72-ab302917'>ab302917</a>) at 1/1000 dilution

Lane 1:

Untreated 3T3-L1 (mouse embryonic fibroblast), whole cell lysate at 50 µg

Lane 2:

3T3-L1 treated with1µM Dexamethasone for 48h, whole cell lysate at 50 µg

Lane 3:

THP-1 (human monocytic leukemia monocyte), whole cell lysate at 50 µg

Lane 4:

Huh7 (Human hepatocellular carcinoma epithelial cell), whole cell lysate at 50 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: 96 kDa

Observed band size: 100-150 kDa

false

• WB

Lab

Western blot - Anti-VLDL Receptor/VLDL-R antibody [EPR26178-72] (BSA and Azide free) (AB302918)

This data was developed using ab302917, the same antibody clone in a different buffer formulation.

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

VLDLR is a glycoprotein of approximately 150 kDa and detected as a 120 kDa band after treated with Protein Deglycosylation MIX II.

Low expression : liver, small intestine (PMID : 7925422; PMID : 11960750; PMID : 29289645)

All lanes:

Western blot - Anti-VLDL Receptor/VLDL-R antibody [EPR26178-72] (<a href='/en-us/products/primary-antibodies/vldl-receptor-vldl-r-antibody-epr26178-72-ab302917'>ab302917</a>) at 1/1000 dilution

Lane 1:

Untreated human heart tissue lysate at 30 µg

Lane 2:

Human heart tissue lysate treated with Protein Deglycosylation Mix II at 30 µg

Lane 3:

Human liver tissue lysate at 30 µg

Lane 4:

Human small intestine tissue lysate at 30 µg

Lane 5:

Untreated mouse heart tissue lysate at 30 µg

Lane 6:

Mouse heart tissue lysate treated with Protein Deglycosylation Mix II at 30 µg

Lane 7:

Mouse liver tissue lysate at 30 µg

Lane 8:

Rat heart tissue lysate at 30 µg

Lane 9:

Rat liver tissue lysate at 30 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/1000 dilution

Predicted band size: 96 kDa

Observed band size: 100-150 kDa

false