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Anti CD9, Rat Monoclonal Antibody(77B), Biotin-conjugated

for Immunochemistry
Manufacturer :
FUJIFILM Wako Pure Chemical Corporation
Storage Condition :
Keep at -20 degrees C.
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If the revision of product standards and packaging standards has been made, there is a case where the actual product specifications and images are different.

SDS

Basic Information

Comparison
Product Number
Package Size
Price
Inventory
Distributor
017-28211
Barcode No
4548995083302
50uL
List Price
418.00 USD

In stock in Japan

Document

SDS
Product Specification Sheet
Package Insert
Spectral Data
Certificate of Analysis
Calibration Certificate

Product Information

Cross Reactivity Human
[Note] This antibody doesn’t recognize the mouse and bovine CD9. It shows a little reactivity with rat CD9.
Clone No. 77B
Host Rat
Subclass IgG2b
Formulation 1xTBS aqueous solution with 50w/v% glycerol, 0.05w/v% sodium azide.
Label Biotin

▼Evaluating the reactivity
The reactivity of this antibody against a serum of each organisms was measured by ELISA.
When the signal value was the same level as the blank, it was judged as “non-reactive”.

Application

ELISA 1:2,000 – 1:16,000
Western Blot 1:1,000 – 1:8,000
* Please use this antibody under non-reducing condition
Immunoprecipitation 1:100 – 1:1,000

Please optimize the most appropriate concentration for your analysis and samples.

Application Data

Characteristic comparison with Clone No. 30B by ELISA

The characteristics of this product and our another anti CD9 antibody were compared by ELISA.

▶The two type of antibodies (77B and 30B) were compared in PS ELISA [our original ELISA kit with Tim4 protein immobilized].

04036523_img01.png

Sample Exosomes from cell culture supernatant of COLO201 cells
ELISA Kit PS Capture™ Exosome ELISA Kit (Streptavidin HRP) [code No. 298-80601]
Detection antibody 77B: Anti CD9, Rat Monoclonal Antibody (77B), Biotin-conjugated [code No. 017-28211] (250 ng/mL)
30B: Anti CD9, Rat Monoclonal Antibody (30B), Biotin-conjugated [code No. 019-27953] (250 ng/mL)

Both the signal value and the S/B ratio of 30B were higher. It has been shown that it is suitable to use 30B as a detection antibody in PS ELISA.

▶Two types of anti-CD9 antibodies (77B and 30B) were used for the immobilization antibody and the detection antibody in the antibody-immobilized ELISA for comparison.

04036523_img02.png
Sample Exosomes from cell culture supernatant of COLO201 cells
Immobilized antibody Anti CD9 Ab: Anti CD9, Rat Monoclonal Antibody (77B) [code No. 019-28173]
Anti CD63 Ab: Anti CD63, Monoclonal Antibody (3-13) [code No. 012-27063]
Anti CD81 Ab: Anti CD81, Monoclonal Antibody (17B1) [code No. 011-27773]
Detection antibody 77B: Anti CD9, Rat Monoclonal Antibody (77B), Biotin-conjugated [code No. 017-28211] (250 ng/mL)
30B: Anti CD9, Rat Monoclonal Antibody (30B), Biotin-conjugated [code No. 019-27953] (250 ng/mL)
Detection HRP-conjugated Streptavidin and TMB solution

Clone No. 30B had higher signal and blank value in antibody-immobilized ELISA, when 30B was used as the detection antibody.
Clone No. 77B had higher S/N ratio than clone No. 30B when compared in terms of S/N ratio.
It has been shown that it is suitable to use 77B as a detection antibody in antibody-immobilized ELISA.

ELISA

Human serum and human heparin plasma were detected by PS ELISA with this antibody.

04036523_img03.png
Sample Human serum and Human heparin plasma [Step Dilution]
ELISA Kit PS Capture™ Exosome ELISA Kit (Streptavidin HRP) [code No. 298-80601]
Detection antibody 77B: Anti CD9, Rat Monoclonal Antibody (77B), Biotin-conjugated [code No. 017-28211] (250 ng/mL)

It was shown that the signal value fluctuated depending on the dilution ratio, and Exosomes in human serum and heparin plasma could be detected.

Western Blot

Isolated exosomes derived from culture supernatant, human serum, and human heparin plasma were electrophoresed under non-reducing conditions, and Western Blot was performed using this antibody.

04036523_img04.png

Sample Exosomes isolated from cell culture supernatant of COLO201 cells, human serum, and human heparin plasma.
Detection antibody Anti CD9, Rat Monoclonal Antibody (77B), Biotin-conjugated [code No. 017-28211] (1 μg/mL)
Detection reagent HRP-Streptavidin, ImmunoStar® Zeta [code No. 291-72401]

CD9 protein (24kDa) in each sample (non-reducing condition) was detected with this antibody.

Immunoprecipitation

The culture supernatant was immunoprecipitated with Anti Rat IgG antibody and this antibody.
Immunoprecipitated samples were analyzed by Western Blot with Anti-CD9 antibody and Anti-Alix antibody (competitors’ products).

04036523_img05.png

Immunoprecipitation
Sample Cell culture supernatant of HEK293T cells
Capture antibody Rat IgG: Biotinylated anti-Rat IgG antibody (as negative control)
77B: Anti CD9, Rat Monoclonal Antibody (77B), Biotin-conjugated [code No. 017-28211]
Immunoprecipitated beads Streptavidin-conjugated magnetic beads
Western Blot
Primary antibody CD9: Anti-CD9 antibody (competitor’s) [1 μg/mL]
Alix: Anti-Alix antibody (competitor’s) [1 μg/mL]
Secondary antibody HRP-conjugated Anti-Mouse IgG antibody
Detection reagent ImmunoStar® Zeta [code No. 291-72401

CD9 and Alix which are marker of exosomes were detected in the sample immunoprecifitated with this antibody.
It was shown that exosomes in samples can be selectively captured by using this product for immunoprecipitation.

Overview / Applications

Outline CD9 is a cell surface glycoprotein which belongs to the tetraspanin superfamily (TM4SF) and, is found on the surface of exosomes. This is known to form a complex with integrins and other TM4SF, can modulate cell adhesion and migration. Also, this induces platelet activation and aggregation.
CD9 is known as exosome marker protein as well as CD63 and CD81.

This antibody is established against human CD9 with Rat, and conjugated with biotin.
It specifically recognizes human CD9 and very slightly rat CD9. It does not recognize bovine and mouse CD9.

[Clone No.] 77B
[Immunized animal] Rat
[Antibody subclass] IgG2b
[Label] Biotin
Application ELISA 1:2000 - 1:16000
Western Blot 1:1000 - 1:8000
*Use this antibody under non-reducing condition.
Immunoprecipitation 1:100 - 1:1000

Please optimize the most appropriate concentration for your analysis and samples.

Property

Specificity Human CD9.
Note: It shows a little reactivity with rat CD9.
It does not react with bovine and mouse CD9.

[Reactivity confirmation condition]
The reactivity of this antibody against the serum of each organisms was measured by ELISA.
When the signal value was the same level as the blank, it was judged as "not react".
Composition 1xTBS, 50w/v% glycerol and 0.05% sodium azide.
Concentration antibody concentration : Indicated on the label.

Manufacturer Information

Alias

Product content may differ from the actual image due to minor specification changes etc.

877-714-1920

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