14-0498-37

antibody from Invitrogen Antibodies

Targeting: ITGA2 CD49B

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Antibody Data

Product number

14-0498-37

Provider

Invitrogen Antibodies

Product name

CD49b (Integrin alpha 2) Monoclonal Antibody (eBioY418 (Y418)), eBioscience™

Antibody type

Monoclonal

Antigen

Other

Description

Description: The eBioY418 (Y418) monoclonal antibody reacts with human CD49b, the 150-kDa, integrin alpha 2 subunit. The complex of CD49b non-covalently associated with integrin beta 1 (CD29), also known as VLA-2, is a receptor for collagen and laminin. This complex is expressed by lymphocytes, activated T cells, monocytes and platelets.

Antibody clone number

eBioY418 (Y418)

Concentration

0.5 mg/mL

References

Submitted references

Prevailing role of contact guidance in intrastromal T-cell trapping in human pancreatic cancer.

Hartmann N, Giese NA, Giese T, Poschke I, Offringa R, Werner J, Ryschich E
Clinical cancer research : an official journal of the American Association for Cancer Research 2014 Jul 1;20(13):3422-33

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot was performed using Anti-CD49b Polyclonal Antibody (Product # 14-0498-80) and a 160kDa band corresponding to CD49b was observed across cell lines tested except U-937. Whole cell extracts (30 µg lysate) of MDA-MB-231 (Lane 1), T-47D (Lane 2), A-431 (Lane 3), U-2 OS (Lane 4), HEK-293 (Lane 5), A549 (Lane 6) and U-937 (Lane 7) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0322BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (0.25 µg/mL) and detected by chemiluminescence Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP (Product # A27036, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005). Note: An uncharacterized band (*) at ~125 kDa was observed in all the samples except U937.

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of CD49b/ITGA2 was performed using 70% confluent log phase U-2 OS cells. The cells were fixed with 4% paraformaldehyde for 10 minutes and blocked with 2% BSA for 1 hour at room temperature. The cells were labeled with CD49b/ITGA2 Mouse Monoclonal Antibody (Product # 14-0498-80) at 5 µg/mL in 0.1% BSA, incubated at 4 degree Celsius overnight and then labeled with Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A28175) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415). Panel d represents the merged image showing membrane localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.