Anti-Tissue Factor antibody [EPR22548-240] - BSA and Azide free

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Tissue Factor antibody [EPR22548-240] - BSA and Azide free (AB254007)

Immunohistochemical analysis of paraffin-embedded human kidney tissue labeling Tissue Factor with ab252918 at 1/2000 dilution, followed by Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining in human renal glomerulus (PMID : 7684196) is observed. Counter stained with hematoxylin.
Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 minutes.

The section was incubated with ab252918 for 30 minutess at RT.

The immunostaining was performed on a Leica Biosystems BOND^® RX instrument.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab252918).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Tissue Factor antibody [EPR22548-240] - BSA and Azide free (AB254007)

Immunohistochemical analysis of paraffin-embedded human cervix carcinoma tissue labeling Tissue Factor with ab252918 at 1/2000 dilution, followed by Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Membranous and weak cytoplasmic staining on tumor cells of human cervix carcinoma (PMID : 26383146) is observed. Counter stained with hematoxylin.
Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 minutes.

The section was incubated with ab252918 for 30 minutes at RT.

The immunostaining was performed on a Leica Biosystems BOND^® RX instrument.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab252918).

• mIHC

Lab

Multiplex immunohistochemistry - Anti-Tissue Factor antibody [EPR22548-240] - BSA and Azide free (AB254007)

Fluorescence multiplex immunohistochemical analysis of paraffin-embedded Human kidney tissue. Panel A : Merged staining of anti-Hexokinase 1 (gray; Opal™690), anti-Angiotensin Converting Enzyme 1 (green; Opal™520) and anti-Tissue Factor (red; Opal™570) on human kidney. Panel B : Anti-Tissue Factor stained on renal glomeruli. Panel C : Anti-Angiotensin Converting Enzyme 1 stained on proximal tubules. Panel D : Anti-Hexokinase 1 stained on distal tubules. The section was incubated in three rounds of staining : in the order of ab150423, ab254222, and ab252918 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system. The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins. Counterstained with DAPI.

• WB

Supplier Data

Western blot - Anti-Tissue Factor antibody [EPR22548-240] - BSA and Azide free (AB254007)

Blocking and dilution buffer : 5% NFDM/TBST.

Exposure times.
Lanes 1 2 : 70 seconds. Lanes 3-6 : 3.25 seconds.

ab252918 was shown to specifically react with Tissue Factor in wild-type HAP1 cells as signal was lost in Tissue Factor knockout cells. Wild-type and Tissue Factor knockout samples were subjected to SDS-PAGE. ab252918 and ab181602 (Rabbit anti-GAPDH loading control) were incubated 1 hour at room temperature at 1/1000 dilution and 1/200,000 dilution respectively. Blots were developed with Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) secondary antibody at 1/100,000 dilution for 1 hour at room temperature before imaging.

The blot was developed on a BIO-RAD ChemiDoc MP instrument using the ECL technique. The molecular weight observed is consistent with what has been described in the literature (PMID : 28938620). Low expression : MCF7 (PMID : 24137414, 28938620).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab252918).

All lanes:

Western blot - Anti-Tissue Factor antibody [EPR22548-240] - BSA and Azide free (ab254007) at 1/1000 dilution

Lane 1:

Wild-type HAP1 whole cell lysate, 60 ug

Lane 2:

Tissue Factor knockout HAP1 whole cell lysate, 60 ug

Lane 3:

A431 (human epidermoid carcinoma cell line) whole cell lysate, 20 ug

Lane 4:

MDA-MB-231 (human breast adenocarcinoma cell line) whole cell lysate, 20 ug

Lane 5:

BxPC-3 (human pancreas adenocarcinoma epithelial cell line) whole cell lysate, 20 ug

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: 33 kDa

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