Revision 9

#97778Store at -20C

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, IHC-Bond, IHC-P, IF-F, IF-IC, FC-FP, FC-L

REACTIVITY:

M R Hm

SENSITIVITY:

Endogenous

MW (kDa):

70-80, 130-140, 200

Source/Isotype:

Rabbit IgG

UniProt ID:

#P31996

Entrez-Gene Id:

12514

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000
IHC Leica Bond 1:600 - 1:2400
Immunohistochemistry (Paraffin) 1:150 - 1:600
Immunofluorescence (Frozen) 1:400 - 1:1600
Immunofluorescence (Immunocytochemistry) 1:200 - 1:400
Flow Cytometry (Fixed/Permeabilized) 1:50 - 1:200
Flow Cytometry (Live) 1:50 - 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.

For a carrier free (BSA and azide free) version of this product see product #29176.

Specificity / Sensitivity

CD68 (E3O7V) Rabbit mAb recognizes endogenous levels of total CD68 protein. Staining of uncertain specificity in mouse endometrial epithelium has been observed by immunohistochemistry.

Species Reactivity:

Mouse, Rat, Hamster

Source / Purification

Monoclonal antibody is produced by immunizing animals with mouse CD68 recombinant protein and reacts with an epitope surrounding Gly113.

Background

CD68 (macrosialin) is a heavily glycosylated transmembrane protein that is expressed by and commonly used as a marker for monocytes and macrophages (1,2). It is found on the plasma membrane, as well as endosomal and lysosomal membranes (1-3). It is proposed to bind OxLDL and has been observed as a homodimer (3,4).

  1. Rabinowitz, S.S. and Gordon, S. (1991) J Exp Med 174, 827-36.
  2. Holness, C.L. and Simmons, D.L. (1993) Blood 81, 1607-13.
  3. Ramprasad, M.P. et al. (1995) Proc Natl Acad Sci U S A 92, 9580-4.
  4. Ramprasad, M.P. et al. (1996) Proc Natl Acad Sci U S A 93, 14833-8.

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 IHC-Bond: IHC Leica Bond IHC-P: Immunohistochemistry (Paraffin) IF-F: Immunofluorescence (Frozen) IF-IC: Immunofluorescence (Immunocytochemistry) FC-FP: Flow Cytometry (Fixed/Permeabilized) FC-L: Flow Cytometry (Live)

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 9
#97778

CD68 (E3O7V) Rabbit mAb

Western Blotting Image 1: CD68 (E3O7V) Rabbit mAb Expand Image
Western blot analysis of extracts from various mouse cells using CD68 (E3O7V) Rabbit mAb (upper) or GAPDH (D16H11) XP® Rabbit mAb #5174 (lower).
Western Blotting Image 2: CD68 (E3O7V) Rabbit mAb Expand Image
Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with a construct expressing Myc/DDK-tagged full-length mouse CD68 protein (mCD68-Myc/DDK; +), using CD68 (E3O7V) Rabbit mAb (upper), Myc-Tag (71D10) Rabbit mAb #2278 (middle), and β-Actin (D6A8) Rabbit mAb #8457 (lower).
Immunohistochemistry Image 1: CD68 (E3O7V) Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded mouse spleen using CD68 (E3O7V) Rabbit mAb performed on the Leica® BOND Rx.
Immunohistochemistry Image 2: CD68 (E3O7V) Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded mouse colon using CD68 (E3O7V) Rabbit mAb performed on the Leica® BOND Rx.
Immunohistochemistry Image 3: CD68 (E3O7V) Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded mouse lung using CD68 (E3O7V) Rabbit mAb performed on the Leica® BOND Rx.
Immunohistochemistry Image 4: CD68 (E3O7V) Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded mouse testis using CD68 (E3O7V) Rabbit mAb performed on the Leica® BOND Rx.
Immunohistochemistry Image 1: CD68 (E3O7V) Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded A20 syngeneic tumor using CD68 (E3O7V) Rabbit mAb.
Immunohistochemistry Image 2: CD68 (E3O7V) Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded 4T1 syngeneic mammary tumor using CD68 (E3O7V) Rabbit mAb.
Immunohistochemistry Image 3: CD68 (E3O7V) Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded Renca syngeneic tumor using CD68 (E3O7V) Rabbit mAb.
Immunohistochemistry Image 4: CD68 (E3O7V) Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded mouse liver using CD68 (E3O7V) Rabbit mAb.
Immunohistochemistry Image 5: CD68 (E3O7V) Rabbit mAb Expand Image
Immunohistochemical anaylsis of paraffin-embedded rat small intestine using CD68 (E3O7V) Rabbit mAb.
Immunohistochemistry Image 6: CD68 (E3O7V) Rabbit mAb Expand Image
Immunohistochemical anaylsis of paraffin-embedded Syrian hamster colon using CD68 (E3O7V) Rabbit mAb.
Immunohistochemistry Image 7: CD68 (E3O7V) Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded mouse small intestine using CD68 (E3O7V) Rabbit mAb (left) compared to concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (right).
Immunohistochemistry Image 8: CD68 (E3O7V) Rabbit mAb Expand Image
Immunohistochemical analysis of paraffin-embedded RAW 264.7 cell pellet (left, positive), Renca cell pellet (middle, positive), or Neuro-2a cell pellet (right, negative) using CD68 (E3O7V) Rabbit mAb.
Immunofluorescence Image 1: CD68 (E3O7V) Rabbit mAb Expand Image
Confocal immunofluorescent analysis of mouse subiculum using CD68 (E3O7V) Rabbit mAb (red). Free secondary binding sites were then blocked with Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 prior to colabeling with GFAP (GA5) Mouse mAb (Alexa Fluor® 488 Conjugate) #3655 (right, green), β-Amyloid (D54D2) XP® Rabbit mAb (Alexa Fluor® 647 Conjugate) #42284 (right, cyan pseudocolor), and DAPI #4083 (right, blue).
Immunofluorescence Image 2: CD68 (E3O7V) Rabbit mAb Expand Image
Confocal immunofluorescent analysis of mouse subiculum from the 5XFAD mouse model of Alzheimer's disease using CD68 (E3O7V) Rabbit mAb (red). Free secondary binding sites were then blocked with Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 prior to colabeling with GFAP (GA5) Mouse mAb (Alexa Fluor® 488 Conjugate) #3655 (right, green), β-Amyloid (D54D2) XP® Rabbit mAb (Alexa Fluor® 647 Conjugate) #42284 (right, cyan pseudocolor), and DAPI #4083 (right, blue).
Immunofluorescence Image 3: CD68 (E3O7V) Rabbit mAb Expand Image
Confocal immunofluorescent analysis of mouse colon (left) and liver (right) using CD68 (E3O7V) Rabbit mAb (red). After blocking free secondary antibody binding sites with Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, the tissue was then labeled using HS1 (D5A9) XP® Rabbit mAb (Rodent Specific) (Alexa Fluor® 488 Conjugate) #68206 (green). Sections were mounted in ProLong® Gold Antifade Reagent with DAPI #8961 (blue).
Immunofluorescence Image 1: CD68 (E3O7V) Rabbit mAb Expand Image
Confocal immunofluorescent analysis of RAW 264.7 cells (left, positive) and Neuro-2a cells (right, negative) using CD68 (E3O7V) Rabbit mAb (green). Cells were mounted in ProLong® Gold Antifade Reagent with DAPI #8961 (blue).
Flow Cytometry Image 1: CD68 (E3O7V) Rabbit mAb Expand Image
Flow cytometric analysis of RAW 264.7 cells, untreated (blue) or treated with TPA (12-O-Tetradecanoylphorbol-13-Acetate) #4174 (100 ng/mL, 24 h; green) using CD68 (E3O7V) Rabbit mAb (solid lines) or 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.