Antibodies, Proteins, Kits and Reagents for Life Science

278 083 ₸ Product size

100 µl 278 083 ₸

Order now and get it on Tuesday March 02, 2021

Anti-Tissue Factor antibody [EPR20369] (ab211016)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR20369] to Tissue Factor
Suitable for: WB, IHC-P
Knockout validated
Reacts with: Human

Product name

Anti-Tissue Factor antibody [EPR20369]
See all Tissue Factor primary antibodies
Description

Rabbit monoclonal [EPR20369] to Tissue Factor
Host species

Rabbit
Tested Applications & Species

Application Species
IHC-P
Human

WB
Human

Recombinant fragment

See all applications and species data
Immunogen

Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: Human Tissue Factor His-tagged recombinant protein (aa34-251); MDA-MB-231 and U-87 MG whole cell lysates; Human fetal kidney and lung cancer lysates. IHC-P: Human placenta. kidney and pancreas cancer tissues.
General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

Application	Species
IHC-P	Human
WB	Human Recombinant fragment

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
Storage buffer

pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR20369
Isotype

IgG
Research areas

Western blot - Anti-Tissue Factor antibody [EPR20369] (ab211016)

All lanes : Anti-Tissue Factor antibody [EPR20369] (ab211016) at 1/1000 dilution (Anti-Tissue Factor antibody NPD)

Lane 1 : Wild-type HAP1 whole cell lysate
Lane 2 : Tissue Factor knockout HAP1 whole cell lysate

Lysates/proteins at 40 µg per lane.

Predicted band size: 33 kDa
Observed band size: 39 kDa
why is the actual band size different from the predicted?

Lanes 1 - 2: Merged signal (red and green). Green - ab211016 observed at 49 kDa. Red - loading control, ab18058, observed at 130 kDa.

ab211016 was shown to recognize Tissue Factor in wild-type HAP1 cells as signal was lost at the expected MW in Tissue Factor knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and F3 (Tissue Factor) knockout samples were subjected to SDS-PAGE. Ab211016 and ab18058 (Mouse anti-Vinculin 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.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Tissue Factor antibody [EPR20369] (ab211016)

Immunohistochemical analysis of paraffin-embedded human kidney tissue labeling Tissue Factor with ab211016 at 1/50 dilution, followed by Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) Ready to use.

Cytoplasmic staining on human kidney glomerulus is observed [PMID: 2237820].

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) Ready to use.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Tissue Factor antibody [EPR20369] (ab211016)

Immunohistochemical analysis of paraffin-embedded human placenta tissue labeling Tissue Factor with ab211016 at 1/50 dilution, followed by Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) Ready to use.

Cytoplasmic staining on human placenta is observed [PMID: 2237820].

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) Ready to use.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Western blot - Anti-Tissue Factor antibody [EPR20369] (ab211016)

Anti-Tissue Factor antibody [EPR20369] (ab211016) at 1/1000 dilution + Human Tissue Factor His-tagged recombinant protein (aa34-251) at 0.01 µg

Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Predicted band size: 33 kDa
Observed band size: 42 kDa why is the actual band size different from the predicted?

Exposure time: 15 seconds

Blocking/Dilution buffer: 5% NFDM/TBST.
Western blot - Anti-Tissue Factor antibody [EPR20369] (ab211016)

All lanes : Anti-Tissue Factor antibody [EPR20369] (ab211016) at 1/5000 dilution

Lane 1 : MDA-MB-231 (Human breast adenocarcinoma cell line) whole cell lysate
Lane 2 : MCF7 (Human breast adenocarcinoma cell line) whole cell lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Predicted band size: 33 kDa
Observed band size: 42 kDa why is the actual band size different from the predicted?

Exposure time: 15 seconds

Blocking/Dilution buffer: 5% NFDM/TBST.

MCF7 is a negative control (PMID: 24137414).
Western blot - Anti-Tissue Factor antibody [EPR20369] (ab211016)

Lanes 1-2 : Anti-Tissue Factor antibody [EPR20369] (ab211016) at 1/1000 dilution
Lane 3 : Anti-Tissue Factor antibody [EPR20369] (ab211016) at 1/10000 dilution

Lane 1 : Human fetal kidney lysate
Lane 2 : Human lung cancer lysate
Lane 3 : U-87 MG (Human glioblastoma-astrocytoma epithelial cell line) whole cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Predicted band size: 33 kDa
Observed band size: 42 kDa why is the actual band size different from the predicted?

Exposure time: 3 minutes

Blocking/Dilution buffer: 5% NFDM/TBST.

The observed molecular weight is consistent with the literature (PMID: 7992837).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Tissue Factor antibody [EPR20369] (ab211016)

Immunohistochemical analysis of paraffin-embedded human pancreas cancer tissue labeling Tissue Factor with ab211016 at 1/50 dilution, followed by Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) Ready to use.

Membranous and cytoplasmic staining on tumor cells of human pancreas cancer is observed [PMID: 7648165].

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) Ready to use.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Anti-Tissue Factor antibody [EPR20369] (ab211016)

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