Antibodies, Proteins, Kits and Reagents for Life Science

Product name

Anti-BAT3/BAG-6 antibody [EPR9223] - BSA and Azide free
See all BAT3/BAG-6 primary antibodies
Description

Rabbit monoclonal [EPR9223] to BAT3/BAG-6 - BSA and Azide free
Host species

Rabbit

Tested Applications & Species

Application	Species
Flow Cyt	Human
IHC-P	Human
WB	Human

See all applications and species data

Immunogen

Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
General notes
ab248802 is the carrier-free version of ab137076 This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with
Ab248802 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

Maxpar® is a trademark of Fluidigm Canada Inc.

This antibody may not be suitable for IHC with mouse or rat samples.
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.
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.

One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.

Learn more here.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C. Do Not Freeze.
Storage buffer

pH: 7.2
Constituent: PBS
Carrier free

Yes
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR9223
Isotype

IgG
Research areas
- Cell Biology
- Apoptosis
- Nucleus
- Other
- Cancer
- Cell Death
- Apoptosis
- Other
- Cancer
- Cell Death
- Apoptosis
- Nucleus
- Other

Western blot - Anti-BAT3/BAG-6 antibody [EPR9223] - BSA and Azide free (ab248802)

This data was developed using ab137076, the same antibody clone in a different buffer formulation.

Lane 1: Wild type HAP1 whole cell lysate (20 µg)

Lane 2: BAT3/BAG-6 knockout HAP1 whole cell lysate (20 µg)

Lane 3: HeLa whole cell lysate (20 µg)

Lanes 1 - 3: Merged signal (red and green). Green - ab137076 observed at 155 kDa. Red - loading control, ab8245, observed at 37 kDa.

ab137076 was shown to specifically react with BAT3/BAG-6 when BAT3/BAG-6 knockout samples were used. Wild-type and BAT3/BAG-6 knockout samples were subjected to SDS-PAGE. ab137076 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/10000 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 1/10000 dilution for 1 hour at room temperature before imaging.
Flow Cytometry - Anti-BAT3/BAG-6 antibody [EPR9223] - BSA and Azide free (ab248802)

This data was developed using ab137076, the same antibody clone in a different buffer formulation.
Overlay histogram showing HAP1 wildtype (green line) and HAP1-BAG6 knockout cells (red line) stained with ab137076. The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Triton X-100 for 15 min. The cells were then incubated in 1 x PBS/10% normal goat serum to block non-specific protein-protein interactions followed by the antibody (ab137076, 0.1µg/ml) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) presorbed (ab150081) at 1/2000 dilution for 30 min at 22°C.

A rabbit IgG isotype control antibody (ab172730) was used at the same concentration and conditions as the primary antibody (HAP1 wildtype - black line, HAP1-BAG6 knockout - grey line). Unlabelled sample was also used as a control (this line is not shown for the purpose of simplicity).

Acquisition of >5,000 events were collected using a 50 mW Blue laser (488nm) and 530/30 bandpass filter.

This antibody can also be used in HAP1 cells fixed with 4% formaldehyde (10 min) permeabilized with 0.1% PBS-Triton X-100 for 15 min under the same conditions.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BAT3/BAG-6 antibody [EPR9223] - BSA and Azide free (ab248802)

This data was developed using ab137076, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of Paraffin-embedded human testis tissue sections labeling BAT3/BAG-6 with purified ab137076 at a dilution of 1/50 dilution (7.1 μg/ml). ab97051 Goat Anti-Rabbit IgG H&L (HRP) at 1/500 was used as the secondary anitbody. Sections were counterstained with hematoxylin. Antigen retrieval was heat mediated using EDTA Buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control and is shown in the inset.
Immunocytochemistry - Anti-BAT3/BAG-6 antibody [EPR9223] - BSA and Azide free (ab248802)

This data was developed using ab137076, the same antibody clone in a different buffer formulation.
Immunocytochemistry/Immunofluorescence analysis of HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling BAT3/BAG-6 with purified ab137076 at 1/500. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. Cells were counter-stained with ab195889 Anti-Alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) at 1/200. DAPI (blue) was used as a nuclear counterstain. Secondary Only Control: PBS was used instead of the primary antibody as the negative control.
Flow Cytometry - Anti-BAT3/BAG-6 antibody [EPR9223] - BSA and Azide free (ab248802)

This data was developed using ab137076, the same antibody clone in a different buffer formulation.
Flow cytometry analysis of HeLa cells labelling BAT3/BAG-6 (red) with purified ab137076 at dilution of 1/30. Goat anti rabbit IgG (Alexa Fluor^®488) at 1/2000 was used as the secondary antibody. Cells were fixed with 4% paraformaldehyde. Isotype control antibody was Rabbit monoclonal IgG (black). The blue line shows cells without incubation with primary antibody and secondary antibody.
Western blot - Anti-BAT3/BAG-6 antibody [EPR9223] - BSA and Azide free (ab248802)

Anti-BAT3/BAG-6 antibody [EPR9223] (ab137076) at 1/5000 dilution (purified) + Mouse brain tissue lysate at 15 µg

Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Predicted band size: 119 kDa

This data was developed using ab137076, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer: 5% NFDM/TBST.
Western blot - Anti-BAT3/BAG-6 antibody [EPR9223] - BSA and Azide free (ab248802)

All lanes : Anti-BAT3/BAG-6 antibody [EPR9223] (ab137076) at 1/1000 dilution (purified)

Lane 1 : A431 (Human epidermoid carcinoma cell line) whole cell lysate
Lane 2 : Human fetal brain tissue lysate
Lane 3 : Rat brain tissue lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/2000 dilution

Predicted band size: 119 kDa
Observed band size: 160 kDa
why is the actual band size different from the predicted?

This data was developed using ab137076, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer: 5% NFDM/TBST.
Flow Cytometry - Anti-BAT3/BAG-6 antibody [EPR9223] - BSA and Azide free (ab248802)

This data was developed using ab137076, the same antibody clone in a different buffer formulation.Overlay histogram showing Hela cells stained with ab137076 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab137076, 1/1000 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1?g/1x10⁶ cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter. This antibody gave a positive signal in HeLa cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BAT3/BAG-6 antibody [EPR9223] - BSA and Azide free (ab248802)

This data was developed using ab137076, the same antibody clone in a different buffer formulation.Immunohistochemical analysis of paraffin-embedded Human kidney tissue labelling BAT3/BAG-6 with unpurified ab137076 at 1/50 dilution.
Western blot - Anti-BAT3/BAG-6 antibody [EPR9223] - BSA and Azide free (ab248802)

All lanes : Anti-BAT3/BAG-6 antibody [EPR9223] (ab137076) at 1/1000 dilution (unpurified)

Lane 1 : HeLa cell lysate
Lane 2 : A431 cell lysate
Lane 3 : Human fetal brain lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : HRP labelled goat anti-rabbit at 1/2000 dilution

Predicted band size: 119 kDa

This data was developed using ab137076, the same antibody clone in a different buffer formulation.
Anti-BAT3/BAG-6 antibody [EPR9223] - BSA and Azide free (ab248802)