Anti-ICAM1 抗体 [EPR4776] - BSA and Azide free

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ICAM1 antibody [EPR4776] - BSA and Azide free (AB226059)

This data was developed using ab109361, the same antibody clone in a different buffer formulation.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human lung carcinoma tissue sections labeling ICAM1 with Purified ab109361 at 1/100 dilution (6.79 µg/mL). Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0). Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Negative control : PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-ICAM1 antibody [EPR4776] - BSA and Azide free (AB226059)

This data was developed using ab109361, the same antibody clone in a different buffer formulation. Intracellular Flow Cytometry analysis of Ramos (Human Burkitt's lymphoma B lymphocyte) cells labeling ICAM1 with Purified ab109361 at 1/70 dilution (10 μg/mL) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-ICAM1 antibody [EPR4776] - BSA and Azide free (AB226059)

This data was developed using ab109361, the same antibody clone in a different buffer formulation.
Immunocytochemistry analysis of Raji (Human Burkitt's lymphoma B lymphocyte) cells labeling ICAM1 with Purified ab109361 at 1/100 dilution (6.79 µg/mL). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1/200 dilution (2.5 µg/mL). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1/1000 dilution (2 µg/mL). DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ICAM1 antibody [EPR4776] - BSA and Azide free (AB226059)

This data was developed using ab109361, the same antibody clone in a different buffer formulation.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human tonsil tissue sections labeling ICAM1 with Purified ab109361 at 1/100 dilution (6.79 µg/mL). Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0). Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Negative control : PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

• IP

Lab

Immunoprecipitation - Anti-ICAM1 antibody [EPR4776] - BSA and Azide free (AB226059)

This data was developed using ab109361, the same antibody clone in a different buffer formulation.
Purified ab109361 at 1/30 dilution (2µg) immunoprecipitating ICAM1 in Raji whole cell lysate.
Lane 1 (input) : Raji (Human Burkitt's lymphoma B lymphocyte) whole cell lysate 10µg
Lane 2 (+) : ab109361 + Raji whole cell lysate.
Lane 3 (-) : Rabbit monoclonal IgG (ab172730) instead of ab109361 in Raji whole cell lysate.
VeriBlot for IP Detection Reagent (HRP) (ab131366) (1/1000 dilution) was used for Western blotting.
Blocking Buffer and concentration : 5% NFDM/TBST.
Diluting buffer and concentration : 5% NFDM/TBST.
Observed band size : 89 kDa

All lanes:

Immunoprecipitation - Anti-ICAM1 antibody [EPR4776] (<a href='/products/primary-antibodies/icam1-antibody-epr4776-ab109361'>ab109361</a>)

Predicted band size: 57 kDa

false

• WB

Lab

Western blot - Anti-ICAM1 antibody [EPR4776] - BSA and Azide free (AB226059)

False colour image of Western blot : Anti-ICAM1 antibody [EPR4776] staining at 1/1000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab109361 was shown to bind specifically to ICAM1. A band was observed at 90 kDa in wild-type HeLa cell lysates with no signal observed at this size in Icam1 knockout cell line ab261742 (knockout cell lysate ab256947). The band observed in the CRISPR-Cas9 edited lysate lane below 90 kDa is likely to represent a truncated form of ICAM1. This has not been investigated further and the functional properties of the gene product have not been determined. To generate this image, wild-type and Icam1 CRISPR-Cas9 edited HeLa cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % 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 IgG H&L (IRDye^® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (ab216776) at 1/20000 dilution.

All lanes:

Western blot - Anti-ICAM1 antibody [EPR4776] (<a href='/products/primary-antibodies/icam1-antibody-epr4776-ab109361'>ab109361</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

ICAM1 CRISPR-Cas9 edited HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human ICAM1 knockout HeLa cell line (<a href='/products/cell-lines/human-icam1-knockout-hela-cell-line-ab261742'>ab261742</a>)

Lane 3:

A549 cell lysate at 20 µg

Lane 4:

Ramos cell lysate at 20 µg

Predicted band size: 57 kDa

Observed band size: 90 kDa

false

• WB

Unknown

Western blot - Anti-ICAM1 antibody [EPR4776] - BSA and Azide free (AB226059)

This data was developed using ab109361, the same antibody clone in a different buffer formulation.

All lanes:

Western blot - Anti-ICAM1 antibody [EPR4776] (<a href='/products/primary-antibodies/icam1-antibody-epr4776-ab109361'>ab109361</a>) at 1/1000 dilution

All lanes:

Ramos (Human Burkitt's lymphoma B lymphocyte) whole cell lysate at 15 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 57 kDa

false

• WB

Lab

Western blot - Anti-ICAM1 antibody [EPR4776] - BSA and Azide free (AB226059)

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

This data was developed using ab109361, the same antibody clone in a different buffer formulation.

All lanes:

Western blot - Anti-ICAM1 antibody [EPR4776] (<a href='/products/primary-antibodies/icam1-antibody-epr4776-ab109361'>ab109361</a>) at 1/1000 dilution

Lane 1:

HUVEC (Human umbilical vein endothelial cell) whole cell lysate at 15 µg

Lane 2:

HUVEC (Human umbilical vein endothelial cell) treated with 10ng/ml TNF-&alpha; for 18 hours whole cell lysate at 15 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 57 kDa

false