Revision 6
Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Applications:

WB, IP, IHC-Bond, IHC-P, IF-F, IF-IC, FC-FP

REACTIVITY:

H M R Mk

SENSITIVITY:

Endogenous

MW (kDa):

120 to 220

Source/Isotype:

Rabbit IgG

UniProt ID:

#P13591

Entrez-Gene Id:

4684

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000
Immunoprecipitation 1:50
IHC Leica Bond 1:50 - 1:200
Immunohistochemistry (Paraffin) 1:100 - 1:400
Immunofluorescence (Frozen) 1:50 - 1:100
Immunofluorescence (Immunocytochemistry) 1:50 - 1:100
Flow Cytometry (Fixed/Permeabilized) 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.

For a carrier free (BSA and azide free) version of this product see product #88856.

Specificity / Sensitivity

NCAM1 (CD56) (E7X9M) XP® Rabbit mAb recognizes endogenous levels of total NCAM1/CD56 protein.

Species Reactivity:

Human, Mouse, Rat, Monkey

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Pro799 of human NCAM1/CD56 protein.

Background

NCAM (neural cell adhesion molecule, CD56) is an adhesion glycoprotein with five extracellular immunoglobulin-like domains followed by two fibronectin type III repeats. Structural diversity is introduced by alternative splicing resulting in different cytoplasmic domains (1). NCAM mediates neuronal attachment, neurite extension, and cell-cell interactions through homo and heterophilic interactions. PSA (polysialic acid) post-translationally modifies NCAM and increases the metastatic potential of small cell lung carcinoma, Wilms' tumor, neuroblastoma, and rhabdomyosarcoma (2). CD56 is commonly used along with CD3 and CD16 to identify human natural killer (NK) cells (mouse NK cells do not express CD56) (3). Human NK cells are CD3-CD56+. The large subset with high CD16 expression are mature cytotoxic NK cells, while those with low CD16 expression are immature precursors and cytokine producers (4,5).

  1. Cunningham, B. A. et al. (1987) Science 236, 799-806.
  2. Seidenfaden, R. et al. (2003) Mol. Cell. Biol. 23, 5908-5918.
  3. Hayakawa, Y. et al. (2006) Immunol Rev 214, 47-55.
  4. Robertson, M.J. and Ritz, J. (1990) Blood 76, 2421-38.
  5. Poli, A. et al. (2009) Immunology 126, 458-65.

Species Reactivity

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

Western Blot Buffer

IMPORTANT: For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.

Applications Key

WB: Western Blotting IP: Immunoprecipitation IHC-Bond: IHC Leica Bond IHC-P: Immunohistochemistry (Paraffin) IF-F: Immunofluorescence (Frozen) IF-IC: Immunofluorescence (Immunocytochemistry) FC-FP: Flow Cytometry (Fixed/Permeabilized)

Cross-Reactivity Key

H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus 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 GP: Guinea Pig Rab: rabbit All: all species expected

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
Alexa Fluor is a registered trademark of Life Technologies Corporation.
All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

使用に関する制限

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Revision 6
#99746

NCAM1 (CD56) (E7X9M) XP® Rabbit mAb

Western Blotting Image 1: NCAM1 (CD56) (E7X9M) XP® Rabbit mAb Expand Image
Western blot analysis of extracts from various cell lines and tissues using NCAM1 (CD56) (E7X9M) XP® Rabbit mAb (upper) or GAPDH (D16H11) XP® Rabbit mAb #5174 (lower).
Immunoprecipitation Image 1: NCAM1 (CD56) (E7X9M) XP® Rabbit mAb Expand Image
Immunoprecipitation of NCAM1 protein from SH-SY5Y cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is NCAM1 (CD56) (E7X9M) XP® Rabbit mAb. Western blot analysis was performed using NCAM1 (CD56) (123C3) Mouse mAb #3576. Anti-mouse IgG, HRP-linked Antibody #7076 was used as the secondary antibody.
Immunohistochemistry Image 1: NCAM1 (CD56) (E7X9M) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human prostate adenocarcinoma with staining of peripheral nerve using NCAM1 (CD56) (E7X9M) XP® Rabbit mAb performed on the Leica® BOND Rx.
Immunohistochemistry Image 1: NCAM1 (CD56) (E7X9M) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human colon adenocarcinoma using NCAM1 (CD56) (E7X9M) XP® Rabbit mAb.
Immunohistochemistry Image 2: NCAM1 (CD56) (E7X9M) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human renal cell carcinoma using NCAM1 (CD56) (E7X9M) XP® Rabbit mAb.
Immunohistochemistry Image 3: NCAM1 (CD56) (E7X9M) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human endometrioid adenocarcinoma using NCAM1 (CD56) (E7X9M) XP® Rabbit mAb.
Immunohistochemistry Image 4: NCAM1 (CD56) (E7X9M) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human gastrointestinal stromal tumor using NCAM1 (CD56) (E7X9M) XP® Rabbit mAb.
Immunohistochemistry Image 5: NCAM1 (CD56) (E7X9M) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded normal rhesus monkey spleen using NCAM1 (CD56) (E7X9M) XP® Rabbit mAb.
Immunohistochemistry Image 6: NCAM1 (CD56) (E7X9M) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded mouse ovary using NCAM1 (CD56) (E7X9M) XP® Rabbit mAb.
Immunohistochemistry Image 7: NCAM1 (CD56) (E7X9M) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded mouse prostate using NCAM1 (CD56) (E7X9M) XP® Rabbit mAb.
Immunohistochemistry Image 8: NCAM1 (CD56) (E7X9M) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded mouse colon using NCAM1 (CD56) (E7X9M) XP® Rabbit mAb.
Immunohistochemistry Image 9: NCAM1 (CD56) (E7X9M) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded endometrioid adenocarcinoma using NCAM1 (CD56) (E7X9M) XP® Rabbit mAb (left) or NCAM (CD56) (123C3) Mouse mAb #3576 (right). These two antibodies detect independent, unique epitopes on human NCAM1. The similar staining patterns obtained with both antibodies help to confirm the specificity of the staining.
Immunohistochemistry Image 10: NCAM1 (CD56) (E7X9M) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded human ductal breast carcinoma using NCAM1 (CD56) (E7X9M) XP® Rabbit mAb (left) compared to concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (right).
Immunohistochemistry Image 11: NCAM1 (CD56) (E7X9M) XP® Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded SH-SY5Y cell pellet (left, positive) or MCF7 cell pellet (right, negative) using NCAM1 (CD56) (E7X9M) XP® Rabbit mAb.
Immunofluorescence Image 1: NCAM1 (CD56) (E7X9M) XP® Rabbit mAb Expand Image
Confocal immunofluorescent analysis of mouse spinal cord (left), or rat brainstem (right) using NCAM1 (CD56) (E7X9M) XP® Rabbit mAb (green) and S6 Ribosomal Protein (54D2) Mouse mAb (Alexa Fluor® 647 Conjugate) #5548 (red). Tissue was mounted in ProLong® Gold Antifade Reagent with DAPI #8961 (blue).
Immunofluorescence Image 1: NCAM1 (CD56) (E7X9M) XP® Rabbit mAb Expand Image
Confocal immunofluorescent analysis of SH-SY5Y cells (left, positive) or HeLa cells (right, negative) using NCAM1 (CD56) (E7X9M) XP® Rabbit mAb (green) and S6 Ribosomal Protein (54D2) Mouse mAb (Alexa Fluor® 647 Conjugate) #5548 (red). Cells were mounted in ProLong® Gold Antifade Reagent with DAPI #8961 (blue).
Immunofluorescence Image 2: NCAM1 (CD56) (E7X9M) XP® Rabbit mAb Expand Image
Confocal immunofluorescent analysis of Neuro-2a cells (left, high-expressing) or C2C12 cells (right, low-expressing) using NCAM1 (CD56) (E7X9M) XP® Rabbit mAb (green) and S6 Ribosomal Protein (54D2) Mouse mAb (Alexa Fluor® 647 Conjugate) #5548 (red). Cells were mounted in ProLong® Gold Antifade Reagent with DAPI #8961 (blue).
Flow Cytometry Image 1: NCAM1 (CD56) (E7X9M) XP® Rabbit mAb Expand Image
Flow cytometric analysis of HeLa cells (blue) and SH-SY5Y cells (green) using NCAM1 (CD56) (E7X9M) XP® 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')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.