Anti-DHFR Antibody Picoband™ (monoclonal, 3C8)

Product Name

Anti-DHFR Antibody Picoband™ (monoclonal, 3C8)

View all Dihydrofolate Reductase/DHFR Antibodies

SKU/Catalog Number

M00813-1

Size

100 μg/vial

Form

Lyophilized

Description

Boster Bio Anti-DHFR Antibody Picoband™ (monoclonal, 3C8) catalog # M00813-1. Tested in IHC, WB applications. This antibody reacts with Human, 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-DHFR Antibody Picoband™ (monoclonal, 3C8) (Boster Biological Technology, Pleasanton CA, USA, Catalog # M00813-1)

Host

Mouse

Contents

Each vial contains 4mg Trehalose, 0.9mg NaCl, 0.2mg Na2HPO4, 0.05mg NaN3.

Clonality

Monoclonal

Clone Number

3C8

Isotype

Mouse IgG2b

Immunogen

E.coli-derived human DHFR recombinant protein (Position: V2-D187). Human DHFRshares90%aminoacid (aa)sequenceidentitywith both mouseandrat DHFR.

*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

M00813-1 is reactive to DHFR in Human, Rat

Applications

M00813-1 is guaranteed for IHC, WB Boster Guarantee

Observed Molecular Weight

22 kDa

Calculated molecular weight

Background of Dihydrofolate Reductase/DHFR

SHC, also known as SHC1 (SHC-transforming protein 1) or SHCA, is a protein that in humans is encoded by the SHC1 gene. SCOP classifies the 3D structure as belonging to the SH2 domain family. This gene encodes three main isoforms that differ in activities and subcellular location. While all three are adapter proteins in signal transduction pathways, the longest (p66Shc) may be involved in regulating life span and the effects of reactive oxygen species. The other two isoforms, p52Shc and p46Shc, link activated receptor tyrosine kinases to the Ras pathway by recruitment of the GRB2/SOS complex. p66Shc is not involved in Ras activation. Unlike the other two isoforms, p46Shc is targeted to the mitochondrial matrix. Several transcript variants encoding different isoforms have been found for this gene.

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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Reconstitution

Add 0.2ml of distilled water will yield a concentration of 500μg/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, 0.1-0.5μg/ml
Immunohistochemistry (Paraffin-embedded Section), 0.5-1μg/ml

Validation Images & Assay Conditions

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Figure 1. Western blot analysis of DHFR using anti-DHFR antibody (M00813-1).
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 SGC-7901 whole cell lysates,
Lane 3: human Hela whole cell lysates,
Lane 4: human SW620 whole cell lysates,
Lane 5: rat kidney 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 mouse anti-DHFR antigen affinity purified monoclonal antibody (Catalog # M00813-1) 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-mouse 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 # EK1001) with Tanon 5200 system. A specific band was detected for DHFR at approximately 22KD. The expected band size for DHFR is at 22KD.

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Figure 2. IHC analysis of DHFR using anti-DHFR antibody (M00813-1).
DHFR was detected in paraffin-embedded section of human rectal cancer 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 mouse anti-DHFR Antibody (M00813-1) overnight at 4°C. Biotinylated goat anti-mouse 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 # SA1021) with DAB as the chromogen.

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Figure 3. IHC analysis of DHFR using anti-DHFR antibody (M00813-1).
DHFR was detected in paraffin-embedded section of rat gaster 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 mouse anti-DHFR Antibody (M00813-1) overnight at 4°C. Biotinylated goat anti-mouse 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 # SA1021) with DAB as the chromogen.

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

Alternative Names

DHFR; DHFRP1; Dihydrofolate Reductase; DYR; EC 1.5.1.3 DHFR DHFRP1, DYR dihydrofolate reductase dihydrofolate reductase

For more info on DHFR, check out the DHFR Infographic

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In this infographic, you will see the following information for DHFR: database IDs, superfamily, protein function, synonyms, molecular weight, chromosomal locations, tissues of expression, subcellular locations, post-translational modifications, and related diseases, research areas & pathways. If you want to see more information included, or would like to contribute to it and be acknowledged, please contact [email protected].

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