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Certificate of Analysis

BDNF Deluxe Research Pack

All You Need for BDNF Research

Cat #: ESD-600_PACK
Size: 16 Vials
Form: Lyophilized

Antibodies

Application key:

CBE- Cell-based ELISA, FC- Flow cytometry, ICC- Immunocytochemistry, IE- Indirect ELISA, IF- Immunofluorescence, IFC- Indirect flow cytometry, IHC- Immunohistochemistry, IP- Immunoprecipitation, LCI- Live cell imaging, N- Neutralization, WB- Western blot

Species reactivity key:

H- Human, M- Mouse, R- Rat

Anti-BDNF Antibody Cat #: ANT-010

Size: 0.2 ml Host: Rabbit Type: Polyclonal Application: IC, IF, IH, WB Reactivity: H, M, R
Immunogen
Peptide (C)VLEKVPVSKGQLK, corresponding to amino acid residues 166-178 of human BDNF (precursor) (Accession P23560).
Homology
Mouse, rat and many other species - identical.

Guinea pig Anti-BDNF Antibody Cat #: ANT-010-GP

Size: 0.2 ml Host: Guinea pig Type: Polyclonal Application: IF, IH, WB Reactivity: H, M, R
Immunogen
Peptide (C)VLEKVPVSKGQLK, corresponding to amino acid residues 166-178 of human BDNF (precursor) (Accession P23560).
Homology
Mouse, rat and many other species - identical.

Anti-p75 NGF Receptor (extracellular) Antibody Cat #: ANT-007

Size: 0.2 ml Host: Rabbit Type: Polyclonal Application: IC, IF, IFC, IH, IP, LCI, WB Reactivity: H, M, R
Immunogen
Peptide CEEIPGRWITRSTPPE, corresponding to amino acid residues 188-203 of human p75NTR (Accession P08138). Extracellular, stalk domain.
Homology
Mouse - identical; rat - 15/16 amino acid residues identical.

Anti-p75 NGF Receptor (extracellular)-ATTO Fluor-550 Antibody Cat #: ANT-007-AO

Size: 50 µl Host: Rabbit Type: Polyclonal Application: IC, IF, IH, LCI Reactivity: H, M, R
Immunogen
Peptide CEEIPGRWITRSTPPE corresponding to amino acid residues 188-203 of human p75NTR (Accession P08138). Extracellular, stalk domain.
Homology
Mouse - identical; rat - 15/16 amino acid residues identical.
Label
ATTO-550. Maximum absorption 554 nm; maximum fluorescence 576 nm. The fluorescence is excited most efficiently in the 540 - 565 nm range. This label is related to the dye Rhodamine 6G and can be used with filters used to detect Rhodamine.

Anti-TrkB (extracellular) Antibody Cat #: ANT-019

Size: 0.2 ml Host: Rabbit Type: Polyclonal Application: IC, IF, IFC, IH, LCI, WB Reactivity: H, M, R
Immunogen
Peptide (C)RLEPNSIDPENITE, corresponding to amino acid residues 57-70 of rat TrkB (Accession Q63604). Extracellular domain.
Homology
Mouse, human - 13/14 amino acid residues identical.

Anti-TrkB (extracellular)-ATTO Fluor-488 Antibody Cat #: ANT-019-AG

Size: 50 µl Host: Rabbit Type: Polyclonal Application: FC, IF, IH Reactivity: H, M, R
Immunogen
Peptide (C)RLEPNSIDPENITE, corresponding to amino acid residues 57-70 of rat TrkB (Accession Q63604). Extracellular domain.
Homology
Mouse, human - 13/14 amino acid residues identical.
Label
ATTO-488. Maximum absorption 501 nm; maximum fluorescence 523 nm. The fluorescence is excited most efficiently in the 480 – 515 nm range. This label is analogous to the well known dye fluorescein isothiocyanate (FITC) and can be used with filters typically used to detect FITC.

Pharmacological Reagents

ANA-12 Cat #: A-215

Size: 10 mg Source: Synthetic MW: 407.49 Purity: >98%
Effective concentration
100-200 µM.
Activity
ANA-12 is a selective TrkB antagonist, shown to bind directly and selectively to TrkB, preventing activation by BDNF and inhibiting processes downstream of TrkB, in a 2-site mode of action (IC50 values are 45.6 nM and 41.1 μM for the high and low affinity sites, respectively), without altering TrkA and TrkC functions. In adult mice ANA-12 decreases TrkB activity in the brain without affecting neuronal survival, and reduces anxiety and depression related behaviors1.
Storage before reconstitution
Shipped at room temperature. Product as supplied can be stored intact at room temperature for several weeks. For longer periods, it should be stored at +4°C.
Reconstitution
Soluble in DMSO. Centrifuge all product preparations before use (10000 x g 5 min).
Storage after reconstitution
Up to four weeks at 4°C or three months at -20°C.

GNF 5837 Cat #: G-145

Size: 25 mg Source: Synthetic MW: 535.49 Purity: >95%
Effective concentration
1 nM - 10 µM.
Activity
GNF 5837 displays antiproliferative effects in cellular Ba/F3 assays (IC50 values are 7, 9 and 11 nM for cells containing the fusion proteins Tel-TrkC, Tel-TrkB and Tel-TrkA, respectively)1.
Storage before reconstitution
Shipped at room temperature. Product as supplied can be stored intact at room temperature for several weeks. For longer periods, it should be stored at -20°C.
Reconstitution
40 mM in DMSO. Centrifuge all product preparations before use (10000 x g for 1 min).
Storage after reconstitution
Up to four weeks at 4°C or three months at -20°C.

Recombinant human BDNF protein Cat #: B-250

Size: 10 µg MW: 27 kDa Purity: >98% (HPLC) Net peptide content: 100%
Origin
Recombinant, E. coli
Effective concentration
ED50 = 220 pM.
Activity
BDNF is a neurotrophic factor and binds p75NTR as well as TrkB receptors1,2. BDNF supports the survival of many cell types3-8.
Storage before reconstitution
The product is shipped as a lyophilized powder at room temperature. Upon receipt, the lyophilizate should be stored at -20°C. Protect from moisture. 
Reconstitution
Sterile water at a concentration of at least 1 µg/0.1 ml. BSA (0.1 mg/ml) should be added for more diluted solutions. Centrifuge all product preparations before use (10000 x g 5 min). Repeated freezing/thawing might result in loss of activity.
Storage after reconstitution
The reconstituted solution can be stored at 4°C for up to 1 week. For longer periods (up to 6 months), small aliquots should be stored at -20°C. We do not recommend storing the product in working solutions for longer than a few days. Avoid multiple freeze-thaw cycles. 

Human TrkB-Fc Chimera Cat #: RPC-001

Size: 50 µg MW: ~120 kDa Purity: >90% (HPLC) Net peptide content: 100%
Origin
Recombinant, HEK 293-6E cells.
Effective concentration
0.1-0.4 µg/ml in the presence of 10 ng/ml of human BDNF.
Activity
TrkB-Fc Chimera  inhibits BDNF-induced phosphorylation of ERK1/2 in HEK293 cells expressing TrkB receptors1-8.
Storage before reconstitution
Shipped at room temperature. Product as supplied can be stored intact at room temperature for several weeks. For longer periods, it should be stored at -20°C.
Reconstitution
Water. Centrifuge all product preparations before use (10000 x g 5 min).
Storage after reconstitution
Up to two weeks at 4°C or three months at -20°C.

Antibody Storage & Reconstitution Instructions

Storage before reconstitution

Lyophilized powder can be stored intact at room temperature for several weeks. For longer periods, it should be stored at -20°C.

Reconstitution

Add deionized water, depending on the sample size.

Storage after reconstitution

The reconstituted solution can be stored at 4°C for up to 2 weeks. For longer periods, small aliquots should be stored at -20°C or below.
Avoid multiple freezing and thawing. The further dilutions should be made using a carrier protein such as BSA (1%). Centrifuge all antibody preparations before use (10000 × g 5 min).

Control antigen storage before reconstitution

Lyophilized powder can be stored intact at room temperature for several weeks. For longer periods, it should be stored at -20°C.

Control antigen reconstitution

100 μl water.

Control antigen storage after reconstitution

-20ºC.

Preadsorption control

1 μg peptide per 1 μg antibody.

For more information, please refer to individual certificate of analysis for each product.
Note
A guinea pig polyclonal antibody (#ANT-010-GP) is included in this Research Pack. Please take into account when reacting with a secondary antibody.
For research purposes only, not for human use