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MTHFD2 (D8W9U) Rabbit mAb

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Product Usage Information

Application Dilution

Western Blotting 1:1000

Immunoprecipitation 1:50

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50%

glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

Specificity / Sensitivity

MTHFD2 (D8W9U) Rabbit mAb recognizes endogenous levels of total MTHFD2 protein.

Species Reactivity:

Human

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Leu340 of human MTHFD2 protein.

Background

MTHFD2 is a bifunctional methylenetetrahydrofolate dehydrogenase/cyclohydrolase involved in mitochondrial folate metabolism (1). MTHFD2 expression is

developmentally regulated, as it is expressed in embryos but not in most adult tissues.

Recent research studies have shown that MTHFD2 is consistently overexpressed in many cancer types and correlated with poor survival in breast cancer (2-5).

Overexpression of MTHFD2 promotes cell proliferation while its depletion induces cell death in human cancer cells (6).

1. Christensen, K.E. and Mackenzie, R.E. (2008) Vitam Horm79, 393-410.

2. Lehtinen, L. et al. (2013) Oncotarget4, 48-63.

3. Nilsson, R. et al. (2014) Nat Commun5, 3128.

4. Liu, F. et al. (2014) Tumour Biol35, 8685-90.

5. Tedeschi, P.M. et al. (2015) Mol Cancer Res13, 1361-6.

6. Gustafsson Sheppard, N. et al. (2015) Sci Rep5, 15029.

For Research Use Only. Not For Use In Diagnostic Procedures.

Applications:

WB, IP

Reactivity:

H

Sensitivity:

Endogenous

MW (kDa):

35

Source/Isotype:

Rabbit IgG

UniProt ID:

P13995

Species reactivity is determined by testing in at least one approved application (e.g., western blot).

IMPORTANT: For primary antibodies recommended for western blotting applications, we recommend incubating the membrane with diluted antibody at 4°C with gentle shaking overnight. Please refer to the western blot protocol found on the product web page for the antibody-specific diluent recommendation.

MTHFD2 (D8W9U) Rabbit mAb

Orders:

Support:

Web:

3 Trask Lane Danvers Massachusetts

(2)

APPLICATIONS KEY WB: Western Blot IP: Immunoprecipitation IHC: Immunohistochemistry ChIP:

Chromatin Immunoprecipitation IF: Immunofluorescence F: Flow Cytometry E-P: ELISA-Peptide CROSS-REACTIVITY KEY H: human M: mouse R: rat Hm: hamster Mk: monkey Mi: mink C:

chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S.

cerevisiae Ce: C. elegans Hr: horse All: all species expected

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

Tween is a registered trademark of ICI Americas, Inc.

Page 2 of 4 MTHFD2 (D8W9U) Rabbit mAb (#41377) Datasheet with Images Cell Signaling Tec...

31/07/2020

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#41377

MTHFD2 (D8W9U) Rabbit mAb

Western blot analysis of extracts from HeLa and 293 cells using MTHFD2 (D8W9U) Rabbit mAb.

(4)

Orders: 877-616-CELL (2355) • [email protected] • Support: 877-678-TECH (8324) • [email protected] • Web:

Page 4 of 4 MTHFD2 (D8W9U) Rabbit mAb (#41377) Datasheet with Images Cell Signaling Tec...

31/07/2020

https://www.cellsignal.com/datasheet.jsp?productId=41377&images=1&protocol=0

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Product Usage Information

Application Dilution

Western Blotting 1:1000

Immunoprecipitation 1:50

Immunohistochemistry (Paraffin) 1:200 - 1:800

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50%

glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

Specificity / Sensitivity

SHMT2 (E7F4Q) Rabbit mAb recognizes endogenous levels of total SHMT2 protein.

This antibody does not cross-react with SHMT1 protein. Species cross-reactivity for IHC-P is human only.

Species Reactivity:

Human, Mouse, Rat, Monkey

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Gly451 of human SHMT2 protein.

Background

Serine hydroxymethyltransferases 1 and 2 (SHMT1, SHMT2) are cytoplasmic and mitochondrial serine hydroxymethyltransferases, respectively (1,2). They catalyze the conversion of serine to glycine with the transfer of β-carbon from serine to

tetrahydrofolate (THF) to form 5, 10-methylene-THF (1, 2). Research studies indicate that SHMT1 hemizygosity is associated with higher risk of intestinal cancer in mice of a certain genetic background (3). Suppression of SHMT2 was shown to block cell proliferation (4).

1. MacFarlane, A.J. et al. (2008) J Biol Chem283, 25846-53.

2. Hebbring, S.J. et al. (2012) J Neurochem120, 881-90.

3. Macfarlane, A.J. et al. (2011) Cancer Res71, 2098-107.

4. di Salvo, M.L. et al. (2013) Med Hypotheses80, 633-6.

For Research Use Only. Not For Use In Diagnostic Procedures.

Applications:

WB, IP, IHC-P

Reactivity:

H M R Mk

Sensitivity:

Endogenous

MW (kDa):

52

Source/Isotype:

Rabbit IgG

UniProt ID:

P34897

Species reactivity is determined by testing in at least one approved application (e.g., western blot).

IMPORTANT: For primary antibodies recommended for western blotting applications, we recommend incubating the membrane with diluted antibody at 4°C with gentle shaking overnight. Please refer to the western blot protocol found on the product web page for the antibody-specific diluent recommendation.

SHMT2 (E7F4Q) Rabbit mAb

Orders:

Support:

Web:

3 Trask Lane Danvers Massachusetts

(6)

APPLICATIONS KEY WB: Western Blot IP: Immunoprecipitation IHC: Immunohistochemistry ChIP:

Chromatin Immunoprecipitation IF: Immunofluorescence F: Flow Cytometry E-P: ELISA-Peptide CROSS-REACTIVITY KEY H: human M: mouse R: rat Hm: hamster Mk: monkey Mi: mink C:

chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S.

cerevisiae Ce: C. elegans Hr: horse All: all species expected

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

SignalStain is a trademark of Cell Signaling Technology, Inc.

XP is a registered trademark of Cell Signaling Technology, Inc.

Tween is a registered trademark of ICI Americas, Inc.

Page 2 of 6 SHMT2 (E7F4Q) Rabbit mAb (#33443) Datasheet with Images Cell Signaling Techn...

31/07/2020

https://www.cellsignal.com/datasheet.jsp?productId=33443&images=1&protocol=0

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#33443

SHMT2 (E7F4Q) Rabbit mAb

Western blot analysis of extracts from RH-30, MDA-MB-231, and HeLa cells using SHMT2 (E7F4Q) Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower).

Immunohistochemical analysis of paraffin-embedded human esophageal carcinoma using SHMT2 (E7F4Q) Rabbit mAb.

Immunoprecipitation of SHMT2 protein from RH-30 cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP Isotype Control #3900, and lane 3 is SHMT2 (E7F4Q) Rabbit mAb. Western blot analysis was performed using SHMT2 (E7F4Q) Rabbit mAb. Mouse Anti-rabbit IgG (Conformation Specific) (L27A9) mAb (HRP Conjugate) #5127 was used as the secondary antibody.

Immunohistochemical analysis of paraffin-embedded human mucoepidermoid carcinoma of the larynx using SHMT2 (E7F4Q) Rabbit mAb.

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Immunohistochemical analysis of paraffin-embedded human colon carcinoma using SHMT2 (E7F4Q) Rabbit mAb.

Immunohistochemical analysis of paraffin-embedded human renal cell carcinoma using SHMT2 (E7F4Q) Rabbit mAb (left) compared to concentration-matched Rabbit (DA1E) mAb IgG XP Isotype Control #3900 (right).®

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Orders: 877-616-CELL (2355) • [email protected] • Support: 877-678-TECH (8324) • [email protected] • Web:

Page 4 of 6 SHMT2 (E7F4Q) Rabbit mAb (#33443) Datasheet with Images Cell Signaling Techn...

31/07/2020

https://www.cellsignal.com/datasheet.jsp?productId=33443&images=1&protocol=0

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#33443

SHMT2 (E7F4Q) Rabbit mAb

Immunohistochemical analysis of paraffin-embedded human non-small cell lung carcinoma using SHMT2 (E7F4Q) Rabbit mAb.

Immunohistochemical analysis of paraffin-embedded RH-30 cell pellet (left, high-expressing) or MDA-MB-231 cell pellet (right, low-expressing) using SHMT2 (E7F4Q) Rabbit mAb.

Immunohistochemical analysis of paraffin-embedded human granulosa cell tumor of the ovary using SHMT2 (E7F4Q) Rabbit mAb.

Immunohistochemical analysis of paraffin-embedded human endometrioid adenocarcinoma using SHMT2 (E7F4Q) Rabbit mAb (left) or SHMT2 (E8O8J) Rabbit mAb (IHC Specific)

#37004 (right). These two antibodies detect independent, unique epitopes on human SHMT2. The similar staining patterns obtained with both antibodies help to confirm the specificity of the staining.

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Orders: 877-616-CELL (2355) • [email protected] • Support: 877-678-TECH (8324) • [email protected] • Web:

Page 6 of 6 SHMT2 (E7F4Q) Rabbit mAb (#33443) Datasheet with Images Cell Signaling Techn...

31/07/2020

https://www.cellsignal.com/datasheet.jsp?productId=33443&images=1&protocol=0

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Product Usage Information

Application Dilution

Western Blotting 1:1000

Immunofluorescence (Immunocytochemistry) 1:800

Flow Cytometry 1:200

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50%

glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

Specificity / Sensitivity

c-Myc (D84C12) Rabbit mAb detects endogenous levels of total c-Myc protein. This antibody is not recommended for detection of Myc-tagged fusion proteins (use Cell Signaling Technology cat. #2276 or #2278).

Species Reactivity:

Human, Mouse, Rat

Species predicted to react based on 100% sequence homology:

Dog, Pig

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to amino-terminal residues of c-Myc.

Background

Members of the Myc/Max/Mad network function as transcriptional regulators with roles in various aspects of cell behavior including proliferation, differentiation and apoptosis (1). These proteins share a common basic-helix-loop-helix leucine zipper (bHLH-ZIP) motif required for dimerization and DNA-binding. Max was originally discovered based on its ability to associate with c-Myc and found to be required for the ability of Myc to bind DNA and activate transcription (2). Subsequently, Max has been viewed as a central component of the transcriptional network, forming homodimers as well as heterodimers with other members of the Myc and Mad families (1). The association between Max and either Myc or Mad can have opposing effects on transcriptional regulation and cell behavior (1). The Mad family consists of four related proteins;

Mad1, Mad2 (Mxi1), Mad3 and Mad4, and the more distantly related members of the bHLH-ZIP family, Mnt and Mga. Like Myc, the Mad proteins are tightly regulated with

short half-lives. In general, Mad family members interfere with Myc-mediated processes such as proliferation, transformation and prevention of apoptosis by inhibiting transcription (3,4).

1. Baudino, T.A. and Cleveland, J.L. (2001) Mol Cell Biol 2. Blackwood, E.M. and Eisenman, R.N. (1991) Science 3. Henriksson, M. and Lüscher, B. (1996) Adv Cancer Res 4. Grandori, C. et al. (2000) Annu Rev Cell Dev Biol16, 653

For Research Use Only. Not For Use In Diagnostic Procedures.

Applications:

WB, IF-IC, F

Reactivity:

H M R

Sensitivity:

Endogenous

MW (kDa):

57-65

Source/Isotype:

Rabbit IgG

UniProt ID:

P01106

Species reactivity is determined by testing in at least one approved application (e.g., western blot).

IMPORTANT: For primary antibodies recommended for western blotting applications, we recommend incubating the membrane with diluted antibody at 4°C with gentle shaking overnight. Please refer to the western blot protocol found on the product web page for the antibody-specific diluent recommendation.

c-Myc (D84C12) Rabbit mAb

Orders:

Support:

Web:

3 Trask Lane Danvers Massachusetts

(12)

APPLICATIONS KEY WB: Western Blot IP: Immunoprecipitation IHC: Immunohistochemistry ChIP:

Chromatin Immunoprecipitation IF: Immunofluorescence F: Flow Cytometry E-P: ELISA-Peptide CROSS-REACTIVITY KEY H: human M: mouse R: rat Hm: hamster Mk: monkey Mi: mink C:

chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S.

cerevisiae Ce: C. elegans Hr: horse All: all species expected

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

Page 2 of 4 c-Myc (D84C12) Rabbit mAb (#5605) Datasheet with Images Cell Signaling Technol...

31/07/2020

https://www.cellsignal.com/datasheet.jsp?productId=5605&images=1&protocol=0

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#5605

c-Myc (D84C12) Rabbit mAb

Western blot analysis of extracts from control HEK293 cells (lane 1) or c-Myc knockout HEK293 cells (lane 2) using c-Myc (D84C12) Rabbit mAb Antibody, #5605 (upper) or β-actin (13E5) Rabbit mAb, #4970 (lower). The absence of signal in the c-Myc knockout HEK293 cells confirms specificity of the antibody for c-Myc.

Confocal immunofluorescent analysis of HeLa cells, mock- transfected (left) or transfected with SignalSilence c-Myc siRNA I #6341 (right), using c-Myc (D84C12) Rabbit mAb (green). Actin filaments have been labeled wth DY-554 phalloidin (red).

®

Western blot analysis of extracts from HeLa cells, mock transfected or transfected with SignalSilence c-Myc siRNA I

#6341, using c-Myc (D84C12) Rabbit mAb.

Flow cytometric analysis of Raji cells using c-Myc (D84C12) Rabbit mAb (blue) compared to a nonspecific negative control antibody (red).

®

Western blot analysis of extracts from various cell lines using c-Myc (D84C12) Rabbit mAb.

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Orders: 877-616-CELL (2355) • [email protected] • Support: 877-678-TECH (8324) • [email protected] • Web:

Page 4 of 4 c-Myc (D84C12) Rabbit mAb (#5605) Datasheet with Images Cell Signaling Technol...

31/07/2020

https://www.cellsignal.com/datasheet.jsp?productId=5605&images=1&protocol=0

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Product Usage Information

For optimal ChIP results, use 10 μl of antibody and 10 μg of chromatin (approximately 4 × 10^6 cells) per IP. This antibody has been validated using SimpleChIP® Enzymatic Chromatin IP Kits

Application Dilution

Western Blotting 1:1000

Immunoprecipitation 1:100

Immunofluorescence (Immunocytochemistry) 1:50 - 1:200

Flow Cytometry 1:200 - 1:800

Chromatin IP 1:100

Chromatin IP-seq 1:100

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50%

glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

Specificity / Sensitivity

c-Myc (E5Q6W) Rabbit mAb recognizes endogenous levels of total c-Myc protein.

Species Reactivity:

Human, Mouse, Rat

Source / Purification

Monoclonal antibody is produced by immunizing animals with recombinant protein specific to the amino terminus of human c-Myc protein.

Background

Members of the Myc/Max/Mad network function as transcriptional regulators with roles in various aspects of cell behavior including proliferation, differentiation and apoptosis (1). These proteins share a common basic-helix-loop-helix leucine zipper (bHLH-ZIP) motif required for dimerization and DNA-binding. Max was originally discovered based on its ability to associate with c-Myc and found to be required for the ability of Myc to bind DNA and activate transcription (2). Subsequently, Max has been viewed as a central component of the transcriptional network, forming homodimers as well as heterodimers with other members of the Myc and Mad families (1). The association between Max and either Myc or Mad can have opposing effects on transcriptional

regulation and cell behavior (1). The Mad family consists of four related proteins;

Mad1, Mad2 (Mxi1), Mad3 and Mad4, and the more distantly related members of the bHLH-ZIP family, Mnt and Mga. Like Myc, the Mad proteins are tightly regulated with short half-lives. In general, Mad family members interfere with Myc-mediated processes such as proliferation, transformation and prevention of apoptosis by inhibiting transcription (3,4).

1. Baudino, T.A. and Cleveland, J.L. (2001) Mol Cell Biol 2. Blackwood, E.M. and Eisenman, R.N. (1991) Science 3. Henriksson, M. and Lüscher, B. (1996) Adv Cancer Res 4. Grandori, C. et al. (2000) Annu Rev Cell Dev Biol16, 653

For Research Use Only. Not For Use In Diagnostic Procedures.

Applications:

WB, IP, IF-IC, F, ChIP, ChIP-seq

Reactivity:

H M R

Sensitivity:

Endogenous

MW (kDa):

57-65

Source/Isotype:

Rabbit IgG

UniProt ID:

P01106

Species reactivity is determined by testing in at least one approved application (e.g., western blot).

IMPORTANT: For primary antibodies recommended for western blotting applications, we recommend incubating the membrane with diluted antibody at 4°C with gentle shaking overnight. Please refer to the western blot protocol found on the product web page for the antibody-specific diluent recommendation.

APPLICATIONS KEY WB: Western Blot IP: Immunoprecipitation IHC: Immunohistochemistry ChIP:

Chromatin Immunoprecipitation IF: Immunofluorescence F: Flow Cytometry E-P: ELISA-Peptide

c-Myc (E5Q6W) Rabbit mAb

Orders:

Support:

Web:

3 Trask Lane Danvers Massachusetts

(16)

CROSS-REACTIVITY KEY H: human M: mouse R: rat Hm: hamster Mk: monkey Mi: mink C:

chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S.

cerevisiae Ce: C. elegans Hr: horse All: all species expected

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

SimpleChIP is a registered trademark of Cell Signaling Technology, Inc.

XP is a registered trademark of Cell Signaling Technology, Inc.

Alexa Fluor is a registered trademark of Life Technologies Corporation.

DyLight is a trademark of Thermo Fisher Scientific, Inc. and its subsidiaries.

Tween is a registered trademark of ICI Americas, Inc.

Page 2 of 4 c-Myc (E5Q6W) Rabbit mAb (#18583) Datasheet with Images Cell Signaling Techno...

31/07/2020

https://www.cellsignal.com/datasheet.jsp?productId=18583&images=1&protocol=0

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#18583

c-Myc (E5Q6W) Rabbit mAb

Western blot analysis of extracts from various cell lines using c-Myc (E5Q6W) Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).

Flow cytometric analysis of IMR-32 cells (blue, negative) and RPMI 8226 cells (green, positive) using c-Myc (E5Q6W) Rabbit mAb (solid lines) or a concentration-matched Rabbit (DA1E) mAb IgG XP Isotype Control #3900 (dashed lines). Anti-rabbit IgG (H+L), F(ab') Fragment (Alexa Fluor 488 Conjugate)

#4412 was used as a secondary antibody.

®

2 ®

Immunoprecipitation of c-Myc from KG-1 cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP Isotype Control #3900, and lane 3 is c-Myc (E5Q6W) Rabbit mAb.

Western blot was performed using c-Myc (E5Q6W) Rabbit mAb. Mouse Anti-rabbit IgG (Conformation Specific) (L27A9) mAb (HRP Conjugate) #5127 was used as a secondary antibody.

Chromatin immunoprecipitations were performed with cross- linked chromatin from NCCIT and either c-Myc (E5Q6W) Rabbit mAb or Normal Rabbit IgG #2729 using SimpleChIP Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. The enriched DNA was quantified by real-time PCR using human ATF-4 Promoter Primers, SimpleChIP Human NPM1 Intron 1 Primers #4779, and SimpleChIP Human αSatellite Repeat Primers #4486. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.

®

®

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®

Confocal immunofluorescent analysis of SCLC-21H cells (left, positive) and IMR-32 cells (right, negative) using c-Myc (E5Q6W) Rabbit mAb (green). Actin filaments were labeled with DyLight™ 554 Phalloidin #13054 (red).

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Orders: 877-616-CELL (2355) • [email protected] • Support: 877-678-TECH (8324) • [email protected] • Web:

Page 4 of 4 c-Myc (E5Q6W) Rabbit mAb (#18583) Datasheet with Images Cell Signaling Techno...

31/07/2020

https://www.cellsignal.com/datasheet.jsp?productId=18583&images=1&protocol=0

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