MOLECULAR SIGNATURE®
Anti-Nitrotyrosine Antibody [39B6]

Mouse Anti- Nitrotyrosine Monoclonal IgG2a

Catalog No. SMC-154

5 out of 5 based on 1 customer rating
Species Reactivity ALL
Applications WB IHC ICC/IF FCM IP
SKU: SMC-154 Categories: ,

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SMC-154_Nitrotyrosine_Antibody_39B6_IHC_Mouse_Retinal-Injury-Model_1.png
Mouse Anti-Nitrotyrosine Antibody [39B6] used in Immunohistochemistry (IHC) on Mouse inflamed colon (SMC-154)Mouse Anti-Nitrotyrosine Antibody [39B6] used in Immunohistochemistry (IHC) on Rat liver tissue  (SMC-154)Mouse Anti-Nitrotyrosine Antibody [39B6] used in Immunohistochemistry (IHC) on Mouse backskin (SMC-154)Mouse Anti-Nitrotyrosine Antibody [39B6] used in Immunohistochemistry (IHC) on Human colon carcinoma (SMC-154)Mouse Anti-Nitrotyrosine Antibody [39B6] used in Western Blot (WB) on Human A549 cells (SMC-154)Mouse Anti-Nitrotyrosine Antibody [39B6] used in Western Blot (WB) on Human Recombinant Protein (SMC-154)
Product Name Nitrotyrosine Antibody
Description

Mouse Anti- Nitrotyrosine Monoclonal IgG2a

Species Reactivity Species Independent
Applications WB, IHC, ICC/IF, IP, ELISA, FCM, AM
Antibody Dilution WB (1:1400), IHC (1:100); optimal dilutions for assays should be determined by the user.
Host Species Mouse
Immunogen 3-(4-hydroxy-3-nitrophenylacetamido) propionic acid-bovine serum albumin
Concentration 1 mg/ml
Conjugates APC, ATTO 390, ATTO 488, ATTO 594, Biotin, FITC, HRP, PerCP, RPE, Unconjugated
Dylight 488
Overview:

  • High fluorescence yield
  • High photostability
  • Less pH-sensitive
  • Excellent batch-to-batch reproducibility
  • Stringently QC tested
  • Molecular weight: 1011 g/mol

Dylight 488 Datasheet

Dylight 488 Fluorophore Absorption and Emission Spectrum Optical Properties:

λex = 493 nm

λem = 518 nm

εmax = 7.0×104

Laser = 488 nm

 

APC/Cy7
Overview:

  • High quantum yield
  • Excellent batch-to-batch reproducibility
  • Stringently QC tested

APC-Cy7 Datasheet

 

ACP-Cy7 Fluorophore Absorption and Emission Spectrum Optical Properties:

λex = 652 nm

λem = 790 nm

Laser = 594 or 633 nm

 

 

  Dylight 350
Overview:

  • High fluorescence intensity
  • High photostability
  • Less pH-sensitive
  • Excellent solubility in water
  • Stringently QC tested
  • Excellent batch-to-batch reproducibility
  • Molecular weight: 874 g/mol

Dylight 350 Datasheet

Dylight 350 Fluorophore Absorption and Emission Spectrum Optical Properties:

λex = 353 nm

λem = 432 nm

εmax = 1.5×104

 

 

  Dylight 405
Overview:

  • High fluorescence intensity
  • High photostability
  • Less pH-sensitive
  • Excellent batch-to-batch reproducibility
  • Stringently QC tested
  • Molecular weight: 793 g/mol

Dylight 405 Datasheet

Dylight 405 Fluorophore Absorption and Emission Spectrum Optical Properties:

λex = 400 nm

λem = 420 nm

εmax = 3.0×104

Laser = 405 nm

 

Dylight 594
Overview:

  • High fluorescence yield
  • High photostability
  • Less pH-sensitive
  • Excellent batch-to-batch reproducibility
  • Stringently QC tested
  • Molecular weight: 1078 g/mol

Dylight 594 Datasheet

Dylight 594 Fluorophore Absorption and Emission Spectrum Optical Properties:

λex = 593 nm

λem = 618 nm

εmax = 8.0×104

Laser = 526 nm

 

 Dylight 633
Overview:

  • High fluorescence yield
  • High photostability
  • Less pH-sensitive
  • Excellent batch-to-batch reproducibility
  • Stringently QC tested
  • Molecular weight: 1066 g/mol

Dylight 633 Datasheet

Dylight 633 Fluorophore Absorption and Emission Spectrum Optical Properties:

λex = 638 nm

λem = 658 nm

εmax = 1.7×105

Laser = 633 nm

 

 PerCP 
Overview:

  • Peridinin-Chlorophyll-Protein Complex
  • Small phycobiliprotein
  • Isolated from red algae
  • Large stokes shift (195 nm)
  • Molecular Weight: 35 kDa

PerCP Datasheet

 PerCP Fluorophore Absorption and Emission Spectrum Optical Properties:

λex = 482 nm

λem = 677 nm

εmax = 1.96 x 106

Laser = 488 nm

 

 PE/ATTO 594
PE/ATTO 594 is a tandem conjugate, where PE is excited at 535 nm and transfers energy to ATTO 594 via FRET (fluorescence resonance energy transfer), which emits at 627 nm.
Overview:

  • High fluorescence yield
  • High photostability
  • Very hydrophilic
  • Excellent solubility in water
  • Very little aggregation

PE/ATTO 594 Datasheet

PE-ATTO 594 Fluorophore Conjugate Excitation and Emission Spectra Optical Properties:

λex = 535 nm

λem = 627 nm

Laser = 488 to 561 nm

 

  FITC (Fluorescein)
Overview:

  • Excellent fluorescence quantum yield
  • High rate of photobleaching
  • Good solubility in water
  • Broad emission spectrum
  • pH dependent spectra
  • Molecular formula: C20H12O5
  • Molar mass: 332.3 g/mol

FITC-Fluorescent-conjugate

FITC Fluorescein Fluorophore Excitation and Emission Spectra Optical Properties:

λex = 494 nm

λem = 520 nm

εmax = 7.3×104

Φf = 0.92

τfl = 5.0 ns

Brightness = 67.2

Laser = 488 nm

Filter set = FITC

 

 ATTO 700
Overview:

  • High fluorescence yield
  • Excellent thermal and photostability
  • Quenched by electron donors
  • Very hydrophilic
  • Good solubility in polar solvents
  • Zwitterionic dye
  • Molar Mass: 575 g/mol

ATTO 700 Datasheet

 ATTO 700 Fluorophore Absorption and Emission Spectrum Optical Properties:

λex = 700 nm

λem = 719 nm

εmax = 1.25×105

Φf = 0.25

τfl = 1.6 ns

Brightness = 31.3

Laser = 676 nm

Filter set = Cy®5.5

 

 ATTO 680
Overview:

  • High fluorescence yield
  • Excellent thermal and photostability
  • Quenched by electron donors
  • Very hydrophilic
  • Good solubility in polar solvents
  • Zwitterionic dye
  • Molar Mass: 631 g/mol

ATTO 680 Datasheet

 ATTO 680 Fluorophore Absorption and Emission Spectrum Optical Properties:

λex = 680 nm

λem = 700 nm

εmax = 1.25×105

Φf = 0.30

τfl = 1.7 ns

Brightness = 37.5

Laser = 633 – 676 nm

Filter set = Cy®5.5

 

 ATTO 655
Overview:

  • High fluorescence yield
  • High thermal and photostability
  • Excellent ozone resistance
  • Quenched by electron donors
  • Very hydrophilic
  • Good solubility in polar solvents
  • Zwitterionic dye
  • Molar Mass: 634 g/mol

ATTO 655 Datasheet

ATTO 655 Fluorophore Absorption and Emission Spectrum Optical Properties:

λex = 663 nm

λem = 684 nm

εmax = 1.25×105

Φf = 0.30

τfl = 1.8 ns

Brightness = 37.5

Laser = 633 – 647 nm

Filter set = Cy®5

 

 ATTO 633
Overview:

  • High fluorescence yield
  • High thermal and photostability
  • Moderately hydrophilic
  • Good solubility in polar solvents
  • Stable at pH 4 – 11
  • Cationic dye, perchlorate salt
  • Molar Mass: 652.2 g/mol

ATTO 633 Datasheet

ATTO 633 Fluorophore Absorption and Emission Spectrum Optical Properties:

λex = 629 nm

λem = 657 nm

εmax = 1.3×105

Φf = 0.64

τfl = 3.2 ns

Brightness = 83.2

Laser = 633 nm

Filter set = Cy®5

 

 ATTO 594
Overview:

  • High fluorescence yield
  • High photostability
  • Very hydrophilic
  • Excellent solubility in water
  • Very little aggregation
  • New dye with net charge of -1
  • Molar Mass: 1137 g/mol

ATTO 594 Datasheet

 ATTO 594 Fluorophore Excitation and Emission Spectrum Optical Properties:

λex = 601 nm

λem = 627 nm

εmax = 1.2×105

Φf = 0.85

τfl = 3.5 ns

Brightness = 102

Laser = 594 nm

Filter set = Texas Red®

 

 ATTO 565
Overview:

  • High fluorescence yield
  • High thermal and photostability
  • Good solubility in polar solvents
  • Excellent solubility in water
  • Very little aggregation
  • Rhodamine dye derivative
  • Molar Mass: 611 g/mol

ATTO 565 Datasheet

 ATTO 565 Fluorophore Excitation and Emission Spectra Optical Properties:

λex = 563 nm

λem = 592 nm

εmax = 1.2×105

Φf = 0.9

τfl = 3.4 n

Brightness = 10

Laser = 532 nm

Filter set = TRITC

 

  ATTO 488
Overview:

  • High fluorescence yield
  • High photostability
  • Very hydrophilic
  • Excellent solubility in water
  • Very little aggregation
  • New dye with net charge of -1
  • Molar Mass: 804 g/mol 

ATTO 488 Datasheet

  ATTO 488 Fluorophore Excitation and Emission Spectra Optical Properties:

λex = 501 nm

λem = 523 nm

εmax = 9.0×104

Φf = 0.80

τfl = 4.1 ns

Brightness = 72

Laser = 488 nm

Filter set = FITC

 

  ATTO 390
Overview:

  • High fluorescence yield
  • Large Stokes-shift (89 nm)
  • Good photostability
  • Moderately hydrophilic
  • Good solubility in polar solvents
  • Coumarin derivate, uncharged
  • Low molar mass: 343.42 g/mol 

ATTO 390 Datasheet

ATTO 390 Fluorescent Dye Excitation and Emission Spectra Optical Properties:

λex = 390 nm

λem = 479 nm

εmax = 2.4×104

Φf = 0.90

τfl = 5.0 ns

Brightness = 21.6

Laser = 365 or 405 nm

 

APC (Allophycocyanin)
Overview:

  • High quantum yield
  • Large phycobiliprotein
  • 6 chromophores per molecule
  • Isolated from red algae
  • Molecular Weight: 105 kDa

APC Datasheet

 APC Fluorophore Absorption and Emission Spectrum Optical Properties:

λex = 650 nm

λem = 660 nm

εmax = 7.0×105

Φf = 0.68

Brightness = 476

Laser = 594 or 633 nm

Filter set = Cy®5

 

Streptavidin

Properties:

  • Homo-tetrameric protein purified from Streptomyces avidinii which binds four biotin molecules with extremely high affinity
  • Molecular weight: 53 kDa
  • Formula: C10H16N2O3S
  • Applications: Western blot, immunohistochemistry, and ELISA

Streptavidin Datasheet

BiotinBiotin Conjugate Structure

Properties:

  • Binds tetrameric avidin proteins including Streptavidin and neuravidin with very high affinity
  • Molar mass: 244.31 g/mol
  • Formula: C10H16N2O3S
  • Applications: Western blot, immunohistochemistry, and ELISA

Biotin Datasheet

HRP (Horseradish peroxidase)

Properties:

  • Enzymatic activity is used to amplify weak signals and increase visibility of a target
  • Readily combines with hydrogen peroxide (H2O2) to form HRP-H2O2 complex which can oxidize various hydrogen donors
  • Catalyzes the conversion of:
    • Chromogenic substrates (e.g. TMB, DAB, ABTS) into coloured products
    • Chemiluminescent substrates (e.g. luminol and isoluminol) into light emitting products via enhanced chemiluminescence (ECL)
    • Fluorogenic substrates (e.g. tyramine, homovanillic acid, and 4-hydroxyphenyl acetic acid) into fluorescent products
  • High turnover rate enables rapid generation of a strong signal
  • 44 kDa glycoprotein
  • Extinction coefficient: 100 (403 nm)
  • Applications: Western blot, immunohistochemistry, and ELISA

HRP Datasheet

AP (Alkaline Phosphatase)

Properties:

  • Broad enzymatic activity for phosphate esters of alcohols, amines, pyrophosphate, and phenols
  • Commonly used to dephosphorylate the 5’-termini of DNA and RNA to prevent self-ligation
  • Catalyzes the conversion of:
    • Chromogenic substrates (e.g. pNPP, naphthol AS-TR phosphate, BCIP) into coloured products
    • Fluorogenic substrates (e.g. 4-methylumbelliferyl phosphate) into fluorescent products
  • Molecular weight: 140 kDa
  • Applications: Western blot, immunohistochemistry, and ELISA

AP Datasheet

  R-PE (R-Phycoerythrin)
Overview:

  • Broad excitation spectrum
  • High quantum yield
  • Photostable
  • Member of the phycobiliprotein family
  • Isolated from red algae
  • Excellent solubility in water
  • Molecular Weight: 250 kDa

R-PE Datasheet

 R-PE Fluorophore Excitation and Emission Spectra Optical Properties:

λex = 565 nm

λem = 575 nm

εmax = 2.0×106

Φf = 0.84

Brightness = 1.68 x 103

Laser = 488 to 561 nm

Filter set = TRITC

 

Field of Use Not for use in humans. Not for use in diagnostics or therapeutics. For in vitro research use only.

Properties

Storage Buffer PBS, 50% glycerol, 0.09% sodium azide *Storage buffer may change when conjugated
Storage Temperature -20ºC, Conjugated antibodies should be stored according to the product label
Shipping Temperature Blue Ice or 4ºC
Purification Protein G Purified
Clonality Monoclonal
Clone Number 39B6
Isotype IgG2a
Specificity Recognizes 3-nitrotyrosine moieties. No detectable cross-reactivity with non-nitrated tyrosine. Not species specific.
Cite This Product StressMarq Biosciences Cat# SMC-154, RRID: AB_904533
Certificate of Analysis 0.7 µg/ml of SMC-154 was sufficient for detection of 5 µg SIN-1 treated BSA by Western Blot analysis using Goat anti-mouse IgG:HRP as the secondary antibody.

Biological Description

Alternative Names Nitro tyrosine Antibody, 3-Nitrotyrosine Antibody
Research Areas Alzheimer's Disease, Cancer, Cell Signaling, Neurodegeneration, Neuroscience, Nitration, Oxidative Stress, Parkinson's Disease, Post-translational Modifications
Scientific Background Protein tyrosine nitration results in a post-translational modification that is increasingly receiving attention as an important component of nitric oxide signaling (2). While multiple nonenzymatic mechanisms are known to be capable of producing nitrated tyrosine residues, most tyrosine nitration events involve catalysis by metalloproteins such as myeloperoxidase, eosino-philperoxidase (3), myoglobin, the cytochrome P-450s, superoxide dismutase and prostacyclin synthase. Nitrotyrosine may also serve as a biomarker for the effects of reactive nitrogen oxides, based on tyrosine residues becoming nitrated in proteins at sites of inflammation induced tissue injury (1). The presence of nitro tyrosine-containing proteins therefore has shown high correlation to disease states such as atherosclerosis, Alzheimer's disease, Parkinson's disease and amyotrophic lateral sclerosis (4).
References 1. Girault I. et al. (2001). Free Radical Biology and Medicine, 31 (11): 1375-1387.
2. Gow AJ, Farkouh CR, Munson DA, Posencheq MA, and Ischiropoulos H. (2004). Am J Physiol Lung Cell Mol Physiol. 287(2): L262-8.
3. Takemoto K. et al (2007). Acta Med Okayama 61(1): 17-30.
4. Reynolds MR. et al. (2006) J Nerosci. 26(42): 10636-45.
5. Pfister H., et al. (2002) Vet Pathol. 39: 190-199.
6. Khan J. et al. (1998) Biochem J. 330(2): 795-801.

Product Images

<p>Immunohistochemistry analysis using Mouse Anti-Nitrotyrosine Monoclonal Antibody, Clone 39B6 (SMC-154). Tissue: Retinal Injury Model. Species: Mouse. Primary Antibody: Mouse Anti-Nitrotyrosine Monoclonal Antibody (SMC-154) at 1:1000. Secondary Antibody: Alexa Fluor 594 Goat Anti-Mouse (red). Courtesy of: Dr. Rajashekhar Gangaraju, University of Indiana, Department of Ophthalmology, Eugene and Marilyn Glick Eye Institute.</p>

Immunohistochemistry analysis using Mouse Anti-Nitrotyrosine Monoclonal Antibody, Clone 39B6 (SMC-154). Tissue: Retinal Injury Model. Species: Mouse. Primary Antibody: Mouse Anti-Nitrotyrosine Monoclonal Antibody (SMC-154) at 1:1000. Secondary Antibody: Alexa Fluor 594 Goat Anti-Mouse (red). Courtesy of: Dr. Rajashekhar Gangaraju, University of Indiana, Department of Ophthalmology, Eugene and Marilyn Glick Eye Institute.

<p>Immunohistochemistry analysis using Mouse Anti-Nitrotyrosine Monoclonal Antibody, Clone 39B6 (SMC-154). Tissue: inflamed colon. Species: Mouse. Fixation: Formalin. Primary Antibody: Mouse Anti-Nitrotyrosine Monoclonal Antibody (SMC-154) at 1:1000000 for 12 hours at 4°C. Secondary Antibody: Biotin Goat Anti-Mouse at 1:2000 for 1 hour at RT. Counterstain: Mayer Hematoxylin (purple/blue) nuclear stain at 200 µl for 2 minutes at RT. Magnification: 40x. With anti-microbial. This image was produced using an amplifying IHC wash buffer. The antibody has therefore been diluted more than is recommended for other applications.</p>

Immunohistochemistry analysis using Mouse Anti-Nitrotyrosine Monoclonal Antibody, Clone 39B6 (SMC-154). Tissue: inflamed colon. Species: Mouse. Fixation: Formalin. Primary Antibody: Mouse Anti-Nitrotyrosine Monoclonal Antibody (SMC-154) at 1:1000000 for 12 hours at 4°C. Secondary Antibody: Biotin Goat Anti-Mouse at 1:2000 for 1 hour at RT. Counterstain: Mayer Hematoxylin (purple/blue) nuclear stain at 200 µl for 2 minutes at RT. Magnification: 40x. With anti-microbial. This image was produced using an amplifying IHC wash buffer. The antibody has therefore been diluted more than is recommended for other applications.

<p>Immunohistochemistry analysis using Mouse Anti-Nitrotyrosine Monoclonal Antibody, Clone 39B6 (SMC-154). Tissue: liver tissue . Species: Rat. Primary Antibody: Mouse Anti-Nitrotyrosine Monoclonal Antibody (SMC-154) at 1:1000. Secondary Antibody: FITC Goat Anti-Mouse (green).</p>

Immunohistochemistry analysis using Mouse Anti-Nitrotyrosine Monoclonal Antibody, Clone 39B6 (SMC-154). Tissue: liver tissue . Species: Rat. Primary Antibody: Mouse Anti-Nitrotyrosine Monoclonal Antibody (SMC-154) at 1:1000. Secondary Antibody: FITC Goat Anti-Mouse (green).

<p>Immunohistochemistry analysis using Mouse Anti-Nitrotyrosine Monoclonal Antibody, Clone 39B6 (SMC-154). Tissue: backskin. Species: Mouse. Fixation: Bouin’s Fixative and paraffin-embedded. Primary Antibody: Mouse Anti-Nitrotyrosine Monoclonal Antibody (SMC-154) at 1:100 for 1 hour at RT. Secondary Antibody: FITC Goat Anti-Mouse (green) at 1:50 for 1 hour at RT. Backskin obtained from transgenic mice.</p>

Immunohistochemistry analysis using Mouse Anti-Nitrotyrosine Monoclonal Antibody, Clone 39B6 (SMC-154). Tissue: backskin. Species: Mouse. Fixation: Bouin’s Fixative and paraffin-embedded. Primary Antibody: Mouse Anti-Nitrotyrosine Monoclonal Antibody (SMC-154) at 1:100 for 1 hour at RT. Secondary Antibody: FITC Goat Anti-Mouse (green) at 1:50 for 1 hour at RT. Backskin obtained from transgenic mice.

<p>Immunohistochemistry analysis using Mouse Anti-Nitrotyrosine Monoclonal Antibody, Clone 39B6 (SMC-154). Tissue: colon carcinoma. Species: Human. Fixation: Formalin. Primary Antibody: Mouse Anti-Nitrotyrosine Monoclonal Antibody (SMC-154) at 1:25000 for 12 hours at 4°C. Secondary Antibody: Biotin Goat Anti-Mouse at 1:2000 for 1 hour at RT. Counterstain: Mayer Hematoxylin (purple/blue) nuclear stain at 200 µl for 2 minutes at RT. Magnification: 40x.</p>

Immunohistochemistry analysis using Mouse Anti-Nitrotyrosine Monoclonal Antibody, Clone 39B6 (SMC-154). Tissue: colon carcinoma. Species: Human. Fixation: Formalin. Primary Antibody: Mouse Anti-Nitrotyrosine Monoclonal Antibody (SMC-154) at 1:25000 for 12 hours at 4°C. Secondary Antibody: Biotin Goat Anti-Mouse at 1:2000 for 1 hour at RT. Counterstain: Mayer Hematoxylin (purple/blue) nuclear stain at 200 µl for 2 minutes at RT. Magnification: 40x.

<p>Western Blot analysis of Human A549 cells showing detection of Multiple Bands Nitrotyrosine protein using Mouse Anti-Nitrotyrosine Monoclonal Antibody, Clone 39B6 (SMC-154). Lane 1: MW ladder. Lane 2: Human A549 Cells 15 ug). Load: 15 ug. Block: 5% Skim Milk Powder in TBST. Primary Antibody: Mouse Anti-Nitrotyrosine Monoclonal Antibody (SMC-154) at 1:1000 for 2.5 hours at RT with shaking . Secondary Antibody: Goat anti-mouse IgG:HRP at 1:1000 for 1 hour at RT with shaking . Color Development: Chemiluminescent for HRP (Moss) for 5 min in RT. Predicted/Observed Size: Multiple Bands.</p>

Western Blot analysis of Human A549 cells showing detection of Multiple Bands Nitrotyrosine protein using Mouse Anti-Nitrotyrosine Monoclonal Antibody, Clone 39B6 (SMC-154). Lane 1: MW ladder. Lane 2: Human A549 Cells 15 ug). Load: 15 ug. Block: 5% Skim Milk Powder in TBST. Primary Antibody: Mouse Anti-Nitrotyrosine Monoclonal Antibody (SMC-154) at 1:1000 for 2.5 hours at RT with shaking . Secondary Antibody: Goat anti-mouse IgG:HRP at 1:1000 for 1 hour at RT with shaking . Color Development: Chemiluminescent for HRP (Moss) for 5 min in RT. Predicted/Observed Size: Multiple Bands.

<p>Western Blot analysis of Human Recombinant Protein showing detection of Multiple Bands Nitrotyrosine protein using Mouse Anti-Nitrotyrosine Monoclonal Antibody, Clone 39B6 (SMC-154). Lane 1: MW Ladder. Lane 2: hASYN Monomer (3.84 ug). Lane 3: Nitrosylated hASYN (3.84 ug).. Block: 5% Skim Milk Powder in TBST. Primary Antibody: Mouse Anti-Nitrotyrosine Monoclonal Antibody (SMC-154) at 1:1000 for 2 hours at RT with shaking . Secondary Antibody: Goat anti-mouse IgG:HRP at 1:4000 for 2 hour at RT with shaking . Color Development: Chemiluminescent for HRP (Moss) for 5 min in RT. Predicted/Observed Size: Multiple Bands.</p>

Western Blot analysis of Human Recombinant Protein showing detection of Multiple Bands Nitrotyrosine protein using Mouse Anti-Nitrotyrosine Monoclonal Antibody, Clone 39B6 (SMC-154). Lane 1: MW Ladder. Lane 2: hASYN Monomer (3.84 ug). Lane 3: Nitrosylated hASYN (3.84 ug).. Block: 5% Skim Milk Powder in TBST. Primary Antibody: Mouse Anti-Nitrotyrosine Monoclonal Antibody (SMC-154) at 1:1000 for 2 hours at RT with shaking . Secondary Antibody: Goat anti-mouse IgG:HRP at 1:4000 for 2 hour at RT with shaking . Color Development: Chemiluminescent for HRP (Moss) for 5 min in RT. Predicted/Observed Size: Multiple Bands.

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