Antibodies, Proteins, Kits and Reagents for Life Science

351 792 ₸ Product size

100 µl 351 792 ₸

Order now and get it on Tuesday March 02, 2021

Anti-Integrin alpha 2 antibody [EPR5788] (ab133557)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR5788] to Integrin alpha 2
Suitable for: Flow Cyt, WB, IHC-P
Knockout validated
Reacts with: Mouse, Human

Product name

Anti-Integrin alpha 2 antibody [EPR5788]
See all Integrin alpha 2 primary antibodies
Description

Rabbit monoclonal [EPR5788] to Integrin alpha 2
Host species

Rabbit

Tested Applications & Species

Application	Species
Flow Cyt	Human
IHC-P	Human
WB	Mouse Human

See all applications and species data

Immunogen

Synthetic peptide within Human Integrin alpha 2 aa 1-100 (extracellular). The exact sequence is proprietary.
Positive control
- WB: T47-D, HEK293, 293T and A431 cell lysates. Human placenta and mouse spleen tissue lysates. IHC-P: Human breast carcinoma and colon tissue. Flow Cyt: A549 and A431 cells.
General notes
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.
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. Stable for 12 months at -20°C.
Dissociation constant (K_D)

K_D = 3.20 x 10 ^-12 M
Learn more about K_D
Storage buffer

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

Protein A purified
Clonality

Monoclonal
Clone number

EPR5788
Isotype

IgG
Research areas

Western blot - Anti-Integrin alpha 2 antibody [EPR5788] (ab133557)

Lane 1: Wild-type HAP1 cell lysate (20 µg)
Lane 2: Integrin alpha 2 knockout HAP1 cell lysate (20 µg)
Lane 3: A431 cell lysate (20 µg)
Lane 4: T47D cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab133557 observed at 165 kDa. Red - loading control, ab8245, observed at 37 kDa.

ab133557 was shown to specifically react with Integrin alpha 2 when Integrin alpha 2 knockout samples were used. Wild-type and Integrin alpha 2 knockout samples were subjected to SDS-PAGE. ab133557 and ab8245 (loading control to GAPDH) were diluted 1/10 000 and 1/2000 respectively and incubated overnight at 4°C. Blots were developed withGoat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10,000 dilution for 1 h at room temperature before imaging.
Western blot - Anti-Integrin alpha 2 antibody [EPR5788] (ab133557)

All lanes : Anti-Integrin alpha 2 antibody [EPR5788] (ab133557) at 1/10000 dilution (purified)

Lane 1 : T47-D cell lysate
Lane 2 : HEK293 cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

Predicted band size: 129 kDa
Observed band size: 150 kDa
why is the actual band size different from the predicted?

Blocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST.
Western blot - Anti-Integrin alpha 2 antibody [EPR5788] (ab133557)

Anti-Integrin alpha 2 antibody [EPR5788] (ab133557) at 1/50000 dilution (purified) + A431 cell lysate at 20 µg

Secondary
Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

Predicted band size: 129 kDa
Observed band size: 150 kDa why is the actual band size different from the predicted?

Blocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST.
Western blot - Anti-Integrin alpha 2 antibody [EPR5788] (ab133557)

Anti-Integrin alpha 2 antibody [EPR5788] (ab133557) at 1/10000 dilution (purified) + Mouse spleen tissue lysate at 20 µg

Secondary
Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

Predicted band size: 129 kDa
Observed band size: 150 kDa why is the actual band size different from the predicted?

Blocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST.
Western blot - Anti-Integrin alpha 2 antibody [EPR5788] (ab133557)

All lanes : Anti-Integrin alpha 2 antibody [EPR5788] (ab133557) at 1/10000 dilution (unpurified)

Lane 1 : T47D lysate
Lane 2 : 293T lysate
Lane 3 : Human platelet lysate
Lane 4 : A431 lysate

Lysates/proteins at 10 µg per lane.

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

Predicted band size: 129 kDa
Observed band size: 150 kDa why is the actual band size different from the predicted?
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Integrin alpha 2 antibody [EPR5788] (ab133557)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human breast carcinoma tissue labelling Integrin alpha 2 with purified ab133557 at 1/500. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Integrin alpha 2 antibody [EPR5788] (ab133557)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human colon tissue labelling Integrin alpha 2 with unpurified ab133557 at 1/250.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Flow Cytometry - Anti-Integrin alpha 2 antibody [EPR5788] (ab133557)

Flow Cytometry analysis of A549 cells labelling Integrin alpha 2 with purified ab133557 at 1/60 (red). Cells were fixed with 2% paraformaldehyde. A FITC-conjugated goat anti-rabbit IgG (1/150) was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG. Blue - Unlabelled control, cells without incubation with primary and secondary antibodies.
Flow Cytometry - Anti-Integrin alpha 2 antibody [EPR5788] (ab133557)

Overlay histogram showing A431 cells stained with unpurified ab133557 (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 (ab133557, 1/1000 dilution) for 30 min at 22°C. The secondary antibody used was a goat anti-rabbit Alexa Fluor^® 488 (IgG; H&L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1μg/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.
OI-RD Scanning - Anti-Integrin alpha 2 antibody [EPR5788] (ab133557)

Equilibrium disassociation constant (K_D)
Learn more about K_D

Click here to learn more about K_D
Anti-Integrin alpha 2 antibody [EPR5788] (ab133557)

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