Biolegend, 328309, Alexa Fluor® 647 anti-human CD49a Antibody, 25tests

CD49a is a 200 kD type I transmembrane glycoprotein also known as α 1 integrin, VLA-1 α chain, or Integrin α 1 . It associates with CD29 (β 1 integrin) to form VLA-1 complex, a collagen IV and alminin-1 receptor. It is expressed on activated T cells, monocytes, NK cells, smooth muscle cells, neuronal cells, fibroblasts, and mesenchymal cells. CD49a is an adhesion molecule and is involved in the regulation of leukocyte migration, T cell proliferation, and cytokine production.
25tests
Verified Reactivity: Human
Reported Reactivity: African Green, Baboon, Cynomolgus, Rhesus
Antibody Type: Monoclonal
Host Species: Mouse
Immunogen: Human CTL line
Formulation: Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide and BSA (origin USA)
Preparation: The antibody was purified by affinity chromatography and conjugated with Alexa Fluor® 647 under optimal conditions.
Concentration: Lot-specific (to obtain lot-specific concentration, please enter the lot number in our Concentration and Expiration Lookup or Certificate of Analysis online tools.)
Storage & Handling: The antibody solution should be stored undiluted between 2°C and 8°C, and protected from prolonged exposure to light. Do not freeze.
Application: FC - Quality tested SB - Reported in the literature, not verified in house
Recommended Usage: Each lot of this antibody is quality control tested by immunofluorescent staining with flow cytometric analysis. For flow cytometric staining, the suggested use of this reagent is 5 µl per million cells in 100 µl staining volume or 5 µl per 100 µl of whole blood. * Alexa Fluor® 647 has a maximum emission of 668 nm when it is excited at 633nm / 635nm. Alexa Fluor® and Pacific Blue™ are trademarks of Life Technologies Corporation.View full statement regarding label licenses
Excitation Laser: Red Laser (633 nm)
Application Notes: Additional reported applications include: immunoprecipitation1, immunohistochemical staining1 of acetone-fixed frozen tissue sections, and spatial biology (IBEX)3,4.
Additional Product Notes: Iterative Bleaching Extended multi-pleXity (IBEX) is a fluorescent imaging technique capable of highly-multiplexed spatial analysis. The method relies on cyclical bleaching of panels of fluorescent antibodies in order to image and analyze many markers over multiple cycles of staining, imaging, and, bleaching. It is a community-developed open-access method developed by the Center for Advanced Tissue Imaging (CAT-I) in the National Institute of Allergy and Infectious Diseases (NIAID, NIH).
Application References(PubMed link indicates BioLegend citation): Hemler ME, et al. 1984. J.Immunol. 132:3011 Hemler ME, et al. 1985. J. Biol. Chem. 260:15246 Radtke AJ, et al. 2020. Proc Natl Acad Sci USA. 117:33455-33465. (SB) PubMed Radtke AJ, et al. 2022. Nat Protoc. 17:378-401. (SB) PubMed
Product Citations: Xu H, et al. 2019. Cell Stem Cell. 24:566. PubMed Fu B et al. 2017. Immunity. 47(6):1100-1113 . PubMed Marquardt N, et al. 2019. Nat Commun. 10:3841. PubMed Miki K, et al. 2021. Nat Commun. 12:3596. PubMed Huang YL, et al. 2020. Elife. 9:00. PubMed Marquardt N, et al. 2015. J Immunol. 194:2467. PubMed
RRID: AB_2129242 (BioLegend Cat. No. 328310)
Structure: Integrin alpha chain family, Type I membrane protein, alpha chain of heterodimeric integrin receptor, 200 kD. Associates with CD29 to form VLA-1 complex
Distribution: Activated T cells, monocytes, NK cells, smooth muscle cells, neuronal cells, fibroblasts, and mesenchymal cells
Function: Adhesion, leukocyte migration
Ligand/Receptor: With integrin β1 (CD29) forms receptor for laminin-1 and collagen IV
Cell Type: Fibroblasts, Mesenchymal cells, Mesenchymal Stem Cells, Monocytes, NK cells, T cells
Biology Area: Immunology, Stem Cells
Molecular Family: Adhesion Molecules, CD Molecules
Antigen References: Zola H, et al. Eds. 2007. Leukocyte and Stromal Cell Molecules:The CD Markers. Wiley-Liss Press. p122 Boiret N, et al. 2005. Exp. Hematol. 33:219
Gene ID: 3672
UniProt: View information about CD49a on UniProt.org
Clone: TS2/7
Regulatory Status: RUO
Other Names: α1 integrin, VLA-1 α chain, Integrin α1 chain, ITGA1
Isotype: Mouse IgG1, κ
Q: If an antibody clone has been previously successfully used in IBEX in one fluorescent format, will other antibody formats work as well?
A: It’s likely that other fluorophore conjugates to the same antibody clone will also be compatible with IBEX using the same sample fixation procedure. Ultimately a directly conjugated antibody’s utility in fluorescent imaging and IBEX may be specific to the sample and microscope being used in the experiment. Some antibody clone conjugates may perform better than others due to performance differences in non-specific binding, fluorophore brightness, and other biochemical properties unique to that conjugate.
Q: Will antibodies my lab is already using for fluorescent or chromogenic IHC work in IBEX?
A: Fundamentally, IBEX as a technique that works much in the same way as single antibody panels or single marker IF/IHC. If you’re already successfully using an antibody clone on a sample of interest, it is likely that clone will have utility in IBEX. It is expected some optimization and testing of different antibody fluorophore conjugates will be required to find a suitable format; however, legacy microscopy techniques like chromogenic IHC on fixed or frozen tissue is an excellent place to start looking for useful antibodies.
Q: Are other fluorophores compatible with IBEX?
A: Over 18 fluorescent formats have been screened for use in IBEX, however, it is likely that other fluorophores are able to be rapidly bleached in IBEX. If a fluorophore format is already suitable for your imaging platform it can be tested for compatibility in IBEX.
Q: The same antibody works in one tissue type but not another. What is happening?
A: Differences in tissue properties may impact both the ability of an antibody to bind its target specifically and impact the ability of a specific fluorophore conjugate to overcome the background fluorescent signal in a given tissue. Secondary stains, as well as testing multiple fluorescent conjugates of the same clone, may help to troubleshoot challenging targets or tissues. Using a reference control tissue may also give confidence in the specificity of your staining.
Q: How can I be sure the staining I’m seeing in my tissue is real?
A: In general, best practices for validating an antibody in traditional chromogenic or fluorescent IHC are applicable to IBEX. Please reference the Nature Methods review on antibody based multiplexed imaging for resources on validating antibodies for IBEX.