Antibodies, Proteins, Kits and Reagents for Life Science

274 732 ₸ Product size

100 µl 274 732 ₸

Order now and get it on Wednesday March 10, 2021

Key features and details

Mouse monoclonal [3F3] to RASSF1a
Suitable for: WB, ICC/IF, Flow Cyt
Reacts with: Mouse, Human
Isotype: IgG1

Product name

Anti-RASSF1a antibody [3F3]
See all RASSF1a primary antibodies
Description

Mouse monoclonal [3F3] to RASSF1a
Host species

Mouse
Specificity

In many cases, the results of staining showed speckled pattern or weak staining in cytoplasm but we couldn’t observe microtubule stained with this antibody. In mitotic cell, we could observe the mitotic spindles stained with this antibody and so thought it was kinetochore stained specifically.

Tested Applications & Species

Application	Species
Flow Cyt	Human
ICC/IF	Human
WB	Human

See all applications and species data

Immunogen

Recombinant human RASSF1A purified from E. coli.
Epitope

ab23950 specifically recognises the C1 domain (52-101aa) of RASSF1a
Positive control
- WB: HeLa lysate, 293cells transfected with HA-tagged RASSF 1A gene ICC: A431 cells
General notes

This product was changed from ascites to tissue culture supernatant on 28/02/19. Please note that the dilutions may need to be adjusted accordingly. If you have any questions, please do not hesitate to contact our scientific support team.
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 short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term.
Storage buffer

pH: 7.40
Preservative: 0.02% Sodium azide
Constituents: PBS, 10% Glycerol
Concentration information loading...
Purity

Protein G purified
Clonality

Monoclonal
Clone number

3F3
Myeloma

Sp2/0
Isotype

IgG1
Light chain type

kappa
Research areas

Immunocytochemistry/ Immunofluorescence - Anti-RASSF1a antibody [3F3] (ab23950)

Immunocytochemistry/ Immunofluorescence analysis of RASSF1a in HeLa cells. The cell was stained with ab23950 (1:100). The secondary antibody (green) was used Alexa Fluor 488. DAPI was stained the cell nucleus (blue).
Western blot - Anti-RASSF1a antibody [3F3] (ab23950)

Anti-RASSF1a antibody [3F3] (ab23950) at 1/500 dilution + HeLa cell lysate at 40 µg

Secondary
Goat anti-mouse secondary antibody conjugated to HRP

Predicted band size: 39 kDa
Western blot - Anti-RASSF1a antibody [3F3] (ab23950)

All lanes : Anti-RASSF1a antibody [3F3] (ab23950) at 1/500 dilution

Lane 1 : 293T cell lysate
Lane 2 : RASSF1A transfected 293T cell lysate

Lysates/proteins at 5 µg per lane.

Secondary
All lanes : goat anti-mouse secondary antibody conjugated to HRP

Predicted band size: 39 kDa
Western blot - Anti-RASSF1a antibody [3F3] (ab23950)

All lanes : Anti-RASSF1a antibody [3F3] (ab23950) at 1/1000 dilution

Lane 1 : Hela (H) cell extract
Lane 2 : Mouse brain (MB) extract
Lane 3 : Mouse liver (ML) extract
Lane 4 : Mouse kidney (MK) extract

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat anti-mouse HRP conjugate

Developed using the ECL technique.

Predicted band size: 39 kDa
Observed band size: 40,43 kDa
why is the actual band size different from the predicted?

Arrows indicate RASSF1A protein (about 40kDa in Hela cells, and 43kDa in other tissues from mice).

This image was generated using the ascites version of the product.
Immunocytochemistry/ Immunofluorescence - Anti-RASSF1a antibody [3F3] (ab23950)

A431 cells were labeled with anti-RASSF1a(clone 3F3) ab23950. Detection was done using a biotinylated secondary antibody and Texas-red conjugated streptavidin.

This image was generated using the ascites version of the product.
Western blot - Anti-RASSF1a antibody [3F3] (ab23950)

All lanes : Anti-RASSF1a antibody [3F3] (ab23950) at 1/1000 dilution

Lane 1 : HEK293 cells
Lane 2 : A549 cells
Lane 3 : HeLa cells

Predicted band size: 39 kDa

Proteins were visualised using a goat anti-mouse secondary antibody conjugation to HRP and a ECL detection system.

This image was generated using the ascites version of the product.
Flow Cytometry - Anti-RASSF1a antibody [3F3] (ab23950)

Overlay histogram showing HeLA cells stained with ab23950 (red line). The cells were fixed with 100% 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 (ab23950, 2µg/1x10⁶ cells) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was Mouse IgG1 [ICIGG1] (ab91353, 2µg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed.

This image was generated using the ascites version of the product.

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