Antibodies, Proteins, Kits and Reagents for Life Science

304 886 ₸ Product size

100 µl 304 886 ₸

Order now and get it on Tuesday March 09, 2021

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR3515] to Bag3
Suitable for: WB, IP
Knockout validated
Reacts with: Mouse, Human

Product name

Anti-Bag3 antibody [EPR3515]
See all Bag3 primary antibodies
Description

Rabbit monoclonal [EPR3515] to Bag3
Host species

Rabbit
Tested Applications & Species

Application Species
IP
Mouse

WB
Human

See all applications and species data
Immunogen

Synthetic peptide within Human Bag3 aa 50-150. The exact sequence is proprietary.
Positive control
- 293T, HeLa, MCF7 and K562 cell lysates. IP: Mouse heart lysate.
General notes
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

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
IP	Mouse
WB	Human

Form

Liquid
Storage instructions

Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid repeated freeze / thaw cycles.
Storage buffer

pH: 7.20
Preservative: 0.05% Sodium azide
Constituents: 0.1% BSA, 40% Glycerol (glycerin, glycerine), 9.85% Tris glycine, 50% Tissue culture supernatant
Concentration information loading...
Purity

Tissue culture supernatant
Clonality

Monoclonal
Clone number

EPR3515
Isotype

IgG
Research areas

Western blot - Anti-Bag3 antibody [EPR3515] (ab92309)

Lane 1: Wild-type HAP1 whole cell lysate (20 µg)
Lane 2: BAG3 knockout HAP1 whole cell lysate (20 µg)
Lane 3: HeLa whole cell lysate (20 µg)
Lane 4: K562 whole cell lysate (20 µg)

Lanes 1 - 4: Merged signal (red and green). Green - ab92309 observed at 80 kDa. Red - loading control, ab18058, observed at 130 kDa.

ab92309 was shown to recognize BAG3 in wild-type cells as signal was lost at the expected MW in BAG3 knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and BAG3 knockout samples were subjected to SDS-PAGE. Ab92309 and ab18058 (Mouse anti-Vinculin loading control) were incubated overnight at 4°C at 1 μg/ml 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.
Immunoprecipitation - Anti-Bag3 antibody [EPR3515] (ab92309)

Purified ab92309 at 1/50 dilution (2µg) immunoprecipitating Bag3 in Mouse heart lysate.
Lane 1 (input): Mouse heart lysate 10µg
Lane 2 (+): ab92309 + Mouse heart lysate.
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab92309 in Mouse heart 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: 80 kDa
Western blot - Anti-Bag3 antibody [EPR3515] (ab92309)

All lanes : Anti-Bag3 antibody [EPR3515] (ab92309) at 1/1000 dilution

Lane 1 : 293T cell
lysate
Lane 2 : HeLa cell
lysate
Lane 3 :
MCF7 cell lysate
Lane 4 : K562 cell lysate

Lysates/proteins at 10 µg per lane.

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

Predicted band size: 62 kDa
Observed band size: 80 kDa
why is the actual band size different from the predicted?
Anti-Bag3 antibody [EPR3515] (ab92309)

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com