Antibodies, Proteins, Kits and Reagents for Life Science

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100 µl 311 587 ₸ 10 µl 123 964 ₸

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Anti-Baf180 antibody [EPR15860] (ab196022)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR15860] to Baf180
Suitable for: Flow Cyt, IHC-P, WB, ICC/IF
Knockout validated
Reacts with: Mouse, Human

Product name

Anti-Baf180 antibody [EPR15860]
See all Baf180 primary antibodies
Description

Rabbit monoclonal [EPR15860] to Baf180
Host species

Rabbit

Tested Applications & Species

Application	Species
Flow Cyt	Human
ICC/IF	Human
IHC-P	Human
WB	Human

See all applications and species data

Immunogen

Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: Human fetal kidney lysate; HeLa nuclear lysate; IHC-P: Human kidney tissue; ICC/IF: HeLa cells.
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.
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

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

EPR15860
Isotype

IgG
Research areas

Western blot - Anti-Baf180 antibody [EPR15860] (ab196022)

Lane 1: Wild-type HAP1 cell lysate (20 µg)
Lane 2: Baf180 knockout HAP1 cell lysate (20 µg)
Lane 3: Human kidney tissue lysate (20 µg)
Lane 4: HeLa cell lysate (20 µg)

Lanes 1 - 4: Merged signal (red and green). Green - ab196022 observed at 193 kDa. Red - loading control, ab18058, observed at 124 kDa.

ab196022 was shown to recognize Baf180 when Baf180 knockout samples were used, along with additional cross-reactive bands. Wild-type and Baf180 knockout samples were subjected to SDS-PAGE. ab196022 and ab18058 (loading control to Vinculin) were diluted 1/5000 and 1/10000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.
Flow Cytometry - Anti-Baf180 antibody [EPR15860] (ab196022)

ab196022 staining Baf180 in the human cell line HeLa (human cervix adenocarcinoma) by flow cytometry. Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. The sample was incubated with the primary antibody at a dilution of 1/250. A goat anti rabbit IgG (Alexa Fluor® 488) at a dilution of 1/2000 was used as the secondary antibody.

Isoytype control: Rabbit monoclonal IgG (Black)

Unlabelled control: Cell without incubation with primary antibody and secondary antibody (Blue)
Western blot - Anti-Baf180 antibody [EPR15860] (ab196022)

Anti-Baf180 antibody [EPR15860] (ab196022) at 1/10000 dilution + Human fetal kidney lysate at 10 µg

Secondary
Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

Predicted band size: 193 kDa
Observed band size: 193,35 kDa
why is the actual band size different from the predicted?

Exposure time: 3 minutes

5% NFDM/TBST: Blocking and diluting buffer.
Western blot - Anti-Baf180 antibody [EPR15860] (ab196022)

Anti-Baf180 antibody [EPR15860] (ab196022) at 1/10000 dilution + HeLa (Human epithelial cells from cervix adenocarcinoma) nuclear lysate at 10 µg

Secondary
Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

Predicted band size: 193 kDa
Observed band size: 193 kDa

Exposure time: 15 seconds

5% NFDM/TBST: Blocking and diluting buffer.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Baf180 antibody [EPR15860] (ab196022)

Immunohistochemical analysis of paraffin-embedded Human kidney tissue labeling Baf180 using ab196022 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nuclear staining on Human kidney tissue is observed. Counter stained with Hematoxylin.

Negative control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunocytochemistry/ Immunofluorescence - Anti-Baf180 antibody [EPR15860] (ab196022)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling Baf180 with ab196022 at 1/100 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500 dilution (green). The nuclear counter stain is DAPI (blue).
Nuclear staining on HeLa cell line is observed.
-ve control - Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500 dilution. The nuclear counter stain is DAPI (blue).
Anti-Baf180 antibody [EPR15860] (ab196022)

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