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Alexa Fluor® 647 anti-Bcl-2 Antibody

Pricing & Availability
Clone
100 (See other available formats)
Regulatory Status
RUO
Other Names
B-Cell CLL/Lymphoma 2 (Bcl-2), Protein Phosphatase 1, Regulatory Subunit 50
Isotype
Mouse IgG1
Ave. Rating
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Product Citations
publications
1_100_AF647_Bcl-2_Antibody_FC_032014
Human peripheral blood lymphocytes were treated with Fixation Buffer (Cat. No. 420801) and Permeabilization Wash Buffer (Cat. No. 421002), then stained with Bcl-2 (clone 100) Alexa Fluor® 647 (filled histogram) or mouse IgG1, κ Alexa Fluor® 647 isotype control (open histogram).
  • 1_100_AF647_Bcl-2_Antibody_FC_032014
    Human peripheral blood lymphocytes were treated with Fixation Buffer (Cat. No. 420801) and Permeabilization Wash Buffer (Cat. No. 421002), then stained with Bcl-2 (clone 100) Alexa Fluor® 647 (filled histogram) or mouse IgG1, κ Alexa Fluor® 647 isotype control (open histogram).
  • 2_1_Human_LN_BCL2_CD3
    Confocal image of human lymph node sample acquired using the IBEX method of highly multiplexed antibody-based imaging: BCL2 (cyan) in Cycle 1, CD3 (purple) in Cycle 4. Tissues were prepared using ~1% (vol/vol) formaldehyde and a detergent. Following fixation, samples are immersed in 30% (wt/vol) sucrose for cryoprotection. Images are courtesy of Drs. Andrea J. Radtke and Ronald N. Germain of the Center for Advanced Tissue Imaging (CAT-I) in the National Institute of Allergy and Infectious Diseases (NIAID, NIH).
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658705 25 tests $124
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658706 100 tests $311
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Description

Bcl-2 is a conserved anti-apoptotic protein that plays important roles in normal immunity. It forms homodimers or heterodimers with other Bcl-2 family members. This protein regulates apoptosis through controlling mitochondrial fusion and fission. Phosphorylation of Bcl-2 has been shown to enhance activity to allow response to extracellular growth-factor-mediated signals. Bcl-2 is also regulated by IRES, miR-15a, miR-16-1 and RNA-BP nucleolin. Chromosome 18q21.3 translocation and overexpression of Bcl-2 is frequently observed in follicular lymphomas and some diffuse large B-cell lymphomas.

Product Details
Technical data sheet

Product Details

Verified Reactivity
Human
Antibody Type
Monoclonal
Host Species
Mouse
Immunogen
Synthetic peptide of amino acids 41-54 of human bcl-2 oncoprotein
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 and expiration, please enter the lot number in our Certificate of Analysis online tool.)
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

ICFC - Quality tested
SB - Reported in the literature, not verified in house
Recommended Usage

Each lot of this antibody is quality control tested by intracellular 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 633 nm / 635 nm.


Alexa Fluor® and Pacific Blue™ are trademarks of Life Technologies Corporation.

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Excitation Laser
Red Laser (633 nm)
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)
  1. Pezzella F, et al. 1993. New Eng. J. Med. 329:690. (IHC)
  2. Pezzella F, et al. 1990. Am. J. Pathol. 137:225. (IHC)
  3. Radtke AJ, et al. 2020. Proc Natl Acad Sci U S A. 117:33455-65. (SB) PubMed
  4. Radtke AJ, et al. 2022. Nat Protoc. 17:378-401. (SB) PubMed
Product Citations
  1. Cabello CM, et al. 2009. Free Radic Biol Med. 220:46. PubMed
  2. Lin JR et al. 2018. eLife. 7 pii: e31657. PubMed
  3. Lin J, Sorger M 2015. Sci Rep. 6: 8390. PubMed
  4. Herrera FG, et al. 2019. Int J Radiat Oncol Biol Phys. 103:320. PubMed
  5. Cai J, et al. 2021. eLife. 10:00. PubMed
RRID
AB_2563279 (BioLegend Cat. No. 658705)
AB_2563279 (BioLegend Cat. No. 658706)

Antigen Details

Structure
Two isoforms, 239 or 205 amino acids in length, with a predicted molecular weight of 26 kD or 22 kD and four ‘BH' domains.
Distribution

Outer mitochondrial membrane, nuclear membrane, endoplasmic reticulum membrane.

Function
Regulates apoptosis through controlling mitochondrial fusion and fission.
Interaction
BAX, BAD, BAK, Bcl-X(L),EI24,APAF1, BBC3, BCL2L1, BNIPL, MRPL41, TP53BP2, FKBP8 BAG1, RAF1, EGLN3, G0S2, and BOP.
Cell Type
B cells
Biology Area
Apoptosis/Tumor Suppressors/Cell Death, Cell Biology, Cell Cycle/DNA Replication, Chromatin Remodeling/Epigenetics, Immunology, Mitochondrial Function, Neuroscience, Signal Transduction, Transcription Factors, Ubiquitin/Protein Degradation
Antigen References

1. Lin P, et al. 2013. Curr. Hematol. Malig. Rep. 8:243.
2. Tomita N. 2011. J. Clin. Exp. Hematop. 51:7.
3. Kelly PN, et al. 2011. Cell Death Differ. 18:1414.
4. Willimott S, et al. 2010. Biochem. Soc. Trans. 38:1571.
5. Rolland SG, et al. 2010. Curr. Opin. Cell Biol. 22:852.

Gene ID
596 View all products for this Gene ID
UniProt
View information about Bcl-2 on UniProt.org

Related FAQs

If an antibody clone has been previously successfully used in IBEX in one fluorescent format, will other antibody formats work as well?

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.

Will antibodies my lab is already using for fluorescent or chromogenic IHC work in IBEX?

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.

Are other fluorophores compatible with IBEX?

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.

The same antibody works in one tissue type but not another. What is happening?

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.

How can I be sure the staining I’m seeing in my tissue is real?

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.

Go To Top Version: 3    Revision Date: 04/18/2022

For Research Use Only. Not for diagnostic or therapeutic use.

 

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This data display is provided for general comparisons between formats.
Your actual data may vary due to variations in samples, target cells, instruments and their settings, staining conditions, and other factors.
If you need assistance with selecting the best format contact our expert technical support team.

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