Anti-CD69 Antibody Picoband™

Product Name

Anti-CD69 Antibody Picoband™

View all CD69 Antibodies

SKU/Catalog Number

A00529-2

Size

100 μg/vial

Form

Lyophilized

Description

Boster Bio Anti-CD69 Antibody Picoband™ catalog # A00529-2. Tested in ELISA, Flow Cytometry, IF, IHC, ICC, WB applications. This antibody reacts with Human, Mouse, Rat.

Storage & Handling

Store at -20˚C for one year from date of receipt. After reconstitution, at 4˚C for one month. It can also be aliquotted and stored frozen at -20˚C for six months. Avoid repeated freeze-thaw cycles.

Cite This Product

Anti-CD69 Antibody Picoband™ (Boster Biological Technology, Pleasanton CA, USA, Catalog # A00529-2)

Host

Rabbit

Contents

Each vial contains 4mg Trehalose, 0.9mg NaCl, 0.2mg Na2HPO4, 0.01mg NaN3.

Clonality

Polyclonal

Isotype

Rabbit IgG

Immunogen

E.coli-derived human CD69 recombinant protein (Position: T101-E148).

*Blocking peptide can be purchased. Costs vary based on immunogen length. Contact us for pricing.

Cross-reactivity

No cross-reactivity with other proteins.

Reactive Species

A00529-2 is reactive to CD69 in Human, Mouse, Rat

Applications

A00529-2 is guaranteed for ELISA, Flow Cytometry, IF, IHC, ICC, WB Boster Guarantee

Observed Molecular Weight

22 kDa

Calculated molecular weight

22.559kDa

Background of CD69

This gene encodes a member of the calcium dependent lectin superfamily of type II transmembrane receptors. Expression of the encoded protein is induced upon activation of T lymphocytes, and may play a role in proliferation. Furthermore, the protein may act to transmit signals in natural killer cells and platelets.

Antibody Validation

Boster validates all antibodies on WB, IHC, ICC, Immunofluorescence, and ELISA with known positive control and negative samples to ensure specificity and high affinity, including thorough antibody incubations.

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Reconsitution

Add 0.2ml of distilled water will yield a concentration of 500ug/ml.

Assay Dilutions Recommendation

The recommendations below provide a starting point for assay optimization. The actual working concentration varies and should be decided by the user.

Western blot, 0.25-0.5μg/ml, Human, Mouse, Rat
Immunohistochemistry (Paraffin-embedded Section), 0.5-1μg/ml, Human
Immunocytochemistry/Immunofluorescence, 4μg/ml, Human
Flow Cytometry, 1-3μg/1x10⁶ cells, Human
Direct ELISA, 0.1-0.5μg/ml, Human

Validation Images & Assay Conditions

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Figure 1. Western blot analysis of CD69 using anti-CD69 antibody (A00529-2).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions.
Lane 1: human K562 whole cell lysates,
Lane 2: human HL-60 whole cell lysates,
Lane 3: human Raji whole cell lysates,
Lane 4: mouse SP2/0 whole cell lysates,
Lane 5: rat thymus tissue lysates,
Lane 6: mouse thymus tissue lysates.
After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-CD69 antigen affinity purified polyclonal antibody (Catalog # A00529-2) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for CD69 at approximately 22KD. The expected band size for CD69 is at 22KD.

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Figure 2. IHC analysis of CD69 using anti-CD69 antibody (A00529-2).
CD69 was detected in paraffin-embedded section of human B lymphocytic tumor tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-CD69 Antibody (A00529-2) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1022) with DAB as the chromogen.

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Figure 3. IF analysis of CD69 using anti-CD69 antibody (A00529-2).
CD69 was detected in immunocytochemical section of K562 cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 4μg/mL rabbit anti-CD69 Antibody (A00529-2) overnight at 4°C. DyLight®594 Conjugated Goat Anti-Rabbit IgG (BA1142) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

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Figure 4. Flow Cytometry analysis of human PBMC cells using anti-CD69 antibody (A00529-2).
Overlay histogram showing human PBMC cells stained with A00529-2 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-CD69 Antibody (A00529-2,1μg/1x10⁶ cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x10⁶) used under the same conditions. Unlabelled sample (Red line) was also used as a control.

Gene/Protein Information For CD69 (Source: Uniprot.org, NCBI)

Alternative Names

activation inducer molecule (AIM/CD69); Activation inducer molecule; AIM; BL-AC/P26; CD69 antigen (p60, early T-cell activation antigen); CD69 antigen; CD69 molecule; CD69; CLEC2CC-type lectin domain family 2 member C; C-type lectin domain family 2, member C; EA1; EA-1; early activation antigen CD69; early lymphocyte activation antigen; Early T-cell activation antigen p60; GP32/28; Leu23; Leukocyte surface antigen Leu-23; MLR-3; p60 CD69 AIM, BL-AC/P26, CLEC2C, EA1, GP32/28, MLR-3 CD69 molecule early activation antigen CD69|C-type lectin domain family 2, member C|CD69 antigen (p60, early T-cell activation antigen)|activation inducer molecule (AIM/CD69)|early T-cell activation antigen p60|early lymphocyte activation antigen|leukocyte surface antigen Leu-23

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